2024-03-29T10:20:36Zhttp://repository.helmholtz-hzi.de/oai/requestoai:repository.helmholtz-hzi.de:10033/85042019-08-30T11:32:38Zcom_10033_620652col_10033_620666
2002-03
urn:hdl:10033/8504
Simultaneous Extraction from Bacterioplankton of Total RNA and DNA Suitable for Quantitative Structure and Function Analyses
Weinbauer, Markus G.
Fritz, Ingo
Wenderoth, Dirk F.
Höfle, Manfred G.
2007-02-19T08:58:53Z
2002-03
2007-02-19T08:58:53Z
2002-03
Applied and Environmental Microbiology 2002 68(3):1082-1087
0099-2240
1098-5336
11872453
10.1128/AEM.68.3.1082-1087.2002
http://hdl.handle.net/10033/8504
123726
en_US
Copyright © 2002, American Society for Microbiology
American Society for Microbiology
oai:repository.helmholtz-hzi.de:10033/85112019-08-30T11:26:40Zcom_10033_620652col_10033_620666
2002-10
urn:hdl:10033/8511
Characterization of a Novel Intracellularly Activated Gene from Salmonella enterica Serovar Typhi
Basso, Holger
Rharbaoui, Faiza
Staendner, Lothar H.
Medina, Eva
García-Del Portillo, Francisco
Guzmán, Carlos A.
2007-02-19T09:25:59Z
2002-10
2007-02-19T09:25:59Z
2002-10
Infection and Immunity 2002 70(10):5404-5411
0019-9567
1098-5522
12228264
10.1128/IAI.70.10.5404-5411.2002
http://hdl.handle.net/10033/8511
128351
en_US
Copyright © 2002, American Society for Microbiology
American Society for Microbiology
oai:repository.helmholtz-hzi.de:10033/85182019-08-30T11:32:38Zcom_10033_620652col_10033_620666
2002-12
urn:hdl:10033/8518
The Viable but Nonculturable State and Starvation Are Different Stress Responses of Enterococcus faecalis, as Determined by Proteome Analysis
Heim, Sabina
Del Mar Lleo, Maria
Bonato, Barbara
Guzman, Carlos A.
Canepari, Pietro
2007-02-19T09:43:06Z
2002-12
2007-02-19T09:43:06Z
2002-12
Journal of Bacteriology 2002 184(23):6739-6745
0021-9193
1098-5530
12426365
10.1128/JB.184.23.6739-6745.2002
http://hdl.handle.net/10033/8518
135411
en_US
Copyright © 2002, American Society for Microbiology
American Society for Microbiology
oai:repository.helmholtz-hzi.de:10033/85192019-08-30T11:32:38Zcom_10033_620652col_10033_620666
1996-01-01
urn:hdl:10033/8519
TRANSFAC: a database on transcription factors and their DNA binding sites.
Wingender, E
Dietze, P
Karas, H
Knüppel, R
2007-02-19T09:44:36Z
1996-01-01
2007-02-19T09:44:36Z
1996-01-01
Nucleic Acids Research 1996 24(1):238-241
0305-1048
1362-4962
8594589
http://hdl.handle.net/10033/8519
145586
en_US
oai:repository.helmholtz-hzi.de:10033/86102019-08-30T11:32:15Zcom_10033_620652col_10033_620666
2003-03
urn:hdl:10033/8610
Isolation of Novel Ultramicrobacteria Classified as Actinobacteria from Five Freshwater Habitats in Europe and Asia
Hahn, Martin W.
Lünsdorf, Heinrich
Wu, Qinglong
Schauer, Michael
Höfle, Manfred G.
Boenigk, Jens
Stadler, Peter
2007-02-20T13:05:38Z
2003-03
2007-02-20T13:05:38Z
2003-03
Applied and Environmental Microbiology 2003 69(3):1442-1451
0099-2240
1098-5336
12620827
10.1128/AEM.69.3.1442-1451.2003
http://hdl.handle.net/10033/8610
150105
en_US
Copyright © 2003, American Society for Microbiology
American Society for Microbiology
oai:repository.helmholtz-hzi.de:10033/85252019-08-30T11:31:47Zcom_10033_620652col_10033_620666
1997-01
urn:hdl:10033/8525
Stable expression of pertussis toxin in Bordetella bronchiseptica under the control of a tightly regulated promoter.
Suarez, A
Staendner, L H
Rohde, Manfred
Piatti, G
Timmis, K N
Guzmán, C A
2007-02-19T10:24:12Z
1997-01
2007-02-19T10:24:12Z
1997-01
Applied and Environmental Microbiology 1997 63(1):122-127
0099-2240
1098-5336
8979346
http://hdl.handle.net/10033/8525
168309
en_US
oai:repository.helmholtz-hzi.de:10033/86162019-08-30T11:32:39Zcom_10033_620652col_10033_620666
1997-11
urn:hdl:10033/8616
Development and application of monoclonal antibodies for in situ detection of indigenous bacterial strains in aquatic ecosystems.
Faude, U C
Höfle, M G
2007-02-20T13:11:06Z
1997-11
2007-02-20T13:11:06Z
1997-11
Applied and Environmental Microbiology 1997 63(11):4534-4542
0099-2240
1098-5336
9361440
http://hdl.handle.net/10033/8616
168773
en_US
oai:repository.helmholtz-hzi.de:10033/85352019-08-30T11:24:30Zcom_10033_620652col_10033_620666
1997-06
urn:hdl:10033/8535
Construction and characterization of a live attenuated vaccine candidate against Shigella dysenteriae type 1.
Klee, S R
Tzschaschel, B D
Fält, I
Kärnell, A
Lindberg, A A
Timmis, K N
Guzmán, C A
2007-02-19T10:40:20Z
1997-06
2007-02-19T10:40:20Z
1997-06
Infection and Immunity 1997 65(6):2112-2118
0019-9567
1098-5522
9169740
http://hdl.handle.net/10033/8535
175292
en_US
oai:repository.helmholtz-hzi.de:10033/85502019-08-30T11:24:30Zcom_10033_620652col_10033_620666
1997-02
urn:hdl:10033/8550
Further characterization and in situ localization of chain-like aggregates of the gliding bacteria Myxococcus fulvus and Myxococcus xanthus.
Freese, A
Reichenbach, H
Lünsdorf, H
2007-02-19T12:14:48Z
1997-02
2007-02-19T12:14:48Z
1997-02
Journal of Bacteriology 1997 179(4):1246-1252
0021-9193
1098-5530
9023208
http://hdl.handle.net/10033/8550
178822
en_US
oai:repository.helmholtz-hzi.de:10033/86052019-08-30T11:26:12Zcom_10033_620652col_10033_620666
1992-10
urn:hdl:10033/8605
Specific lung mucosal and systemic immune responses after oral immunization of mice with Salmonella typhimurium aroA, Salmonella typhi Ty21a, and invasive Escherichia coli expressing recombinant pertussis toxin S1 subunit.
Walker, M J
Rohde, Manfred
Timmis, K N
Guzmán, C A
Images
2007-02-20T12:57:13Z
1992-10
2007-02-20T12:57:13Z
1992-10
Infection and Immunity 1992 60(10):4260-4268
0019-9567
1098-5522
1398937
http://hdl.handle.net/10033/8605
257461
en_US
oai:repository.helmholtz-hzi.de:10033/86222019-08-30T11:26:12Zcom_10033_620652col_10033_620666
1991-12
urn:hdl:10033/8622
Antibody responses in the lungs of mice following oral immunization with Salmonella typhimurium aroA and invasive Escherichia coli strains expressing the filamentous hemagglutinin of Bordetella pertussis.
Guzmán, C A
Brownlie, R M
Kadurugamuwa, J
Walker, M J
Timmis, K N
Images
2007-02-20T13:17:22Z
1991-12
2007-02-20T13:17:22Z
1991-12
Infection and Immunity 1991 59(12):4391-4397
0019-9567
1098-5522
1937797
http://hdl.handle.net/10033/8622
259054
en_US
oai:repository.helmholtz-hzi.de:10033/86302019-08-30T11:25:43Zcom_10033_620652col_10033_620666
1994-12
urn:hdl:10033/8630
Invasion and intracellular survival of Bordetella bronchiseptica in mouse dendritic cells.
Guzman, C A
Rohde, Manfred
Bock, M
Timmis, K N
Images
2007-02-20T13:28:24Z
1994-12
2007-02-20T13:28:24Z
1994-12
Infection and Immunity 1994 62(12):5528-5537
0019-9567
1098-5522
7960135
http://hdl.handle.net/10033/8630
303298
en_US
oai:repository.helmholtz-hzi.de:10033/86312019-08-30T11:27:16Zcom_10033_620652col_10033_620666
1994-12
urn:hdl:10033/8631
Mechanisms involved in uptake of Bordetella bronchiseptica by mouse dendritic cells.
Guzman, C A
Rohde, Manfred
Timmis, K N
Images
2007-02-20T13:28:45Z
1994-12
2007-02-20T13:28:45Z
1994-12
Infection and Immunity 1994 62(12):5538-5544
0019-9567
1098-5522
7960136
http://hdl.handle.net/10033/8631
303299
en_US
oai:repository.helmholtz-hzi.de:10033/86612019-08-30T11:24:26Zcom_10033_620652col_10033_620666
2004-03
urn:hdl:10033/8661
Hepatitis C Virus Isolates from Argentina Disclose a Novel Genotype 1-Associated Restriction Pattern
Gismondi, María Inés
Staendner, Lothar Heinrich
Grinstein, Saúl
Guzmán, Carlos Alberto
Preciado, María Victoria
2007-02-20T14:34:20Z
2004-03
2007-02-20T14:34:20Z
2004-03
Journal of Clinical Microbiology 2004 42(3):1298-1301
0095-1137
1098-660X
15004102
10.1128/JCM.42.3.1298-1301.2004
http://hdl.handle.net/10033/8661
356882
en_US
Copyright © 2004, American Society for Microbiology
American Society for Microbiology
oai:repository.helmholtz-hzi.de:10033/86692019-08-30T11:24:26Zcom_10033_620652col_10033_620666
2004-06
urn:hdl:10033/8669
Establishment of a Real-Time PCR-Based Approach for Accurate Quantification of Bacterial RNA Targets in Water, Using Salmonella as a Model Organism
Fey, Axel
Eichler, Stefan
Flavier, Sébastien
Christen, Richard
Höfle, Manfred G.
Guzmán, Carlos A.
2007-02-20T14:39:26Z
2004-06
2007-02-20T14:39:26Z
2004-06
Applied and Environmental Microbiology 2004 70(6):3618-3623
0099-2240
1098-5336
15184165
10.1128/AEM.70.6.3618-3623.2004
http://hdl.handle.net/10033/8669
427797
en_US
Copyright © 2004, American Society for Microbiology
American Society for Microbiology
oai:repository.helmholtz-hzi.de:10033/87512019-08-30T11:25:09Zcom_10033_620652col_10033_620666
2006-02
urn:hdl:10033/8751
Identification of a Thiomicrospira denitrificans-Like Epsilonproteobacterium as a Catalyst for Autotrophic Denitrification in the Central Baltic Sea†
Brettar, Ingrid
Labrenz, Matthias
Flavier, Sébastien
Bötel, Julia
Kuosa, Harri
Christen, Richard
Höfle, Manfred G.
2007-02-22T14:50:22Z
2006-02
2007-02-22T14:50:22Z
2006-02
Applied and Environmental Microbiology 2006 72(2):1364-1372
0099-2240
1098-5336
16461688
10.1128/AEM.72.2.1364-1372.2006
http://hdl.handle.net/10033/8751
1392969
en_US
Copyright © 2006, American Society for Microbiology
American Society for Microbiology
oai:repository.helmholtz-hzi.de:10033/87842019-08-30T11:32:15Zcom_10033_620652col_10033_620666
2006-06
urn:hdl:10033/8784
Phylogenetic Analysis of Previously Nontypeable Hepatitis C Virus Isolates from Argentina†
Gismondi, María Inés
Becker, Pablo Daniel
Valva, Pamela
Guzmán, Carlos Alberto
Preciado, María Victoria
2007-02-22T16:02:01Z
2006-06
2007-02-22T16:02:01Z
2006-06
Journal of Clinical Microbiology 2006 44(6):2229-2232
0095-1137
1098-660X
16757625
10.1128/JCM.02569-05
http://hdl.handle.net/10033/8784
1489453
en_US
Copyright © 2006, American Society for Microbiology
American Society for Microbiology
oai:repository.helmholtz-hzi.de:10033/124722019-08-30T11:37:00Zcom_10033_620652col_10033_620666
2007-07-02T14:45:20Z
urn:hdl:10033/12472
The response of Vibrio- and Rhodobacter-related populations of the NW Mediterranean Sea to additions of dissolved organic matter, phages, or dilution.
Weinbauer, Markus G
Christen, Richard
Höfle, Manfred G
We investigated the growth response of the heterotrophic prokaryotic community focusing on Vibrio- and Rhodobacter-related populations (SRF3) to variation in the availability of dissolved organic matter (DOM), population density-dependent effects, and prokaryotic virus (phage) infection in coastal and offshore waters of the NW Mediterranean Sea. We tested the response of the prokaryotic community to three different DOM fractions prepared by ultrafiltration. One of the DOM fractions contained phages (<0.2 m), a second was virus-free (<100 kDa), and a third contained only low molecular weight (<1 kDa). The proportion of Vibrio and SRF3 populations as determined by fluorescent in situ hybridization in the community ranged from <1 to 6.2% and from 3.2 to 6.3%, respectively. Based on changes in cell numbers, growth rates ranged from 2.1 to 3.1 day(-1) for Vibrio and from 0.8 to 1.2 day(-1) for SRF3. Growth rates of Vibrio were similar or higher than those of the total prokaryotic community, whereas the ability of Vibrio to use high molecular weight (HMW) DOM and the responses to additions of phage-rich material were lower. Growth rates of SRF3 were lower than that of the community. Susceptibility to infection of SRF3 was sometimes lower than in the community, whereas the growth stimulation of HMW DOM was similar or lower. Reducing the cell concentrations of the prokaryotic community by dilution stimulated the overall growth of the community, including that of its constituent Vibrio and SRF3 populations, but the effect was smaller on the SRF3 and greater on Vibrio populations than for the total community. Comparisons with the community also revealed that life strategy traits of bacterial populations differed between coastal and offshore waters. Overall, our data suggest that Vibrio is an r-strategist or opportunistic population in the NW Mediterranean Sea, whereas SRF3 is a K-strategist or equilibrium population.
2007-07-02T14:45:20Z
2007-07-02T14:45:20Z
2006-04-01
Article
Microb. Ecol. 2006, 51(3):336-44
0095-3628
16598637
10.1007/s00248-006-9028-x
http://hdl.handle.net/10033/12472
en
oai:repository.helmholtz-hzi.de:10033/133712019-08-30T11:37:00Zcom_10033_620652col_10033_620666
2007-08-27T14:01:46Z
urn:hdl:10033/13371
Rheinheimera perlucida sp. nov., a marine bacterium of the Gammaproteobacteria isolated from surface water of the central Baltic Sea.
Brettar, Ingrid
Christen, Richard
Höfle, Manfred G
A bacterial isolate from the Baltic Sea, BA131(T), was characterized for its physiological and biochemical features, fatty acid profile, G+C content and phylogenetic position based on comparative 16S rRNA gene sequence analysis. The strain was isolated from surface water of the central Baltic Sea during the decay of a plankton bloom. Phylogenetic analyses of the 16S rRNA gene sequence revealed a clear affiliation with the Gammaproteobacteria, and showed closest phylogenetic relationships with the genera Alishewanella and Rheinheimera. The G+C content of the DNA of strain BA131(T) was 48.9 mol%. Cells were non-pigmented, Gram-negative, rod-shaped, motile by means of a single polar flagellum and catalase- and oxidase-positive. Growth was observed at salinities from 0 to 8 %, with an optimum at 1-3 %. Temperature for growth ranged from 4 to 37 degrees C, with an optimum around 25 degrees C. The fatty acids were dominated by 16 : 0 (17-18 %) and by unsaturated compounds (>61 % of the total): 16 : 1omega7c (24-33 %), 17 : 1omega8c (14-18 %) and 18 : 1omega7c (9-12 %). Based on the data presented, BA131(T) is proposed as the type strain of a novel species of the genus Rheinheimera, Rheinheimera perlucida sp. nov. The type strain is BA131(T) (=LMG 23581(T)=CIP 109200(T)).
2007-08-27T14:01:46Z
2007-08-27T14:01:46Z
2006-09-01
Article
Int. J. Syst. Evol. Microbiol. 2006, 56(Pt 9):2177-83
1466-5026
16957117
10.1099/ijs.0.64172-0
http://hdl.handle.net/10033/13371
en
oai:repository.helmholtz-hzi.de:10033/156762019-08-30T11:25:43Zcom_10033_620652col_10033_620666
2008-01-04T15:13:11Z
urn:hdl:10033/15676
Quantitative reverse transcription polymerase chain reaction analysis of Vibrio cholerae cells entering the viable but non-culturable state and starvation in response to cold shock.
González-Escalona, Narjol
Fey, Axel
Höfle, Manfred G
Espejo, Romilio T
A Guzmán, Carlos
Vaccine Research Group, Division of Microbiology, GBF-German Research Centre for Biotechnology, Braunschweig, Germany.
We performed a comparative analysis of the Vibrio cholerae strain El Tor 3083 entering the viable but non-culturable (VBNC) state and starvation after incubation in artificial seawater (ASW) at 4 and 15 degrees C respectively. To this end, we determined bacterial culturability and membrane integrity, as well as the cellular levels of 16S rRNA and mRNA for the tuf, rpoS and relA genes, which were assessed by real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR). Bacterial cells entering the VBNC state showed a 154, 5.1 x 10(3), 24- and 23-fold reduction in the number of copies of 16S rRNA and mRNA for tuf, rpoS and relA, in comparison to exponentially growing cells. The differences were less striking between cells in the VBNC and starvation states. The mRNA for relA was selectively increased in VBNC cells (3.2-folds), whereas a 3.9-fold reduction was observed for 16S rRNA. The obtained results confirmed that key activities of the cellular metabolism (i.e. tuf representing protein synthesis, and relA or rpoS stress response) were still detected in bacteria entering the VBNC state and starvation. These data suggest that the new Q-RT-PCR methodology, based on the selected RNA targets, could be successfully exploited for the identification (rRNA) of V. cholerae and assessment of its metabolic activity (tuf, rpoS, relA mRNA) in environmental samples.
2008-01-04T15:13:11Z
2008-01-04T15:13:11Z
2006-04
Article
Quantitative reverse transcription polymerase chain reaction analysis of Vibrio cholerae cells entering the viable but non-culturable state and starvation in response to cold shock. 2006, 8 (4):658-66 Environ. Microbiol.
1462-2912
16584477
10.1111/j.1462-2920.2005.00943.x
http://hdl.handle.net/10033/15676
Environmental microbiology
en
oai:repository.helmholtz-hzi.de:10033/158552019-08-30T11:33:29Zcom_10033_620652col_10033_620666
2008-01-09T10:03:02Z
urn:hdl:10033/15855
Vaccines against typhoid fever.
Guzman, Carlos A
Borsutzky, Stefan
Griot-Wenk, Monika
Metcalfe, Ian C
Pearman, Jon
Collioud, Andre
Favre, Didier
Dietrich, Guido
Vaccine Research Group, Division of Microbiology, GBF-German Research Centre for Biotechnology, Mascheroder Weg 1, Braunschweig, Germany.
Because of high infectivity and significant disease burden, typhoid fever constitutes a major global health problem. Implementation of adequate food handling practices and establishment of safe water supplies are the cornerstone for the development of an effective prevention program. However, vaccination against typhoid fever remains an essential tool for the effective management of this disease. Currently, there are two well tolerated and effective licensed vaccines. One is based on defined subunit virulence (Vi) polysaccharide antigen and can be administered either intramuscularly or subcutaneously and the other is based on the use of live attenuated bacteria for oral administration. The advantages and disadvantages of the various approaches taken in the development of a vaccine against typhoid fever are discussed, along with the potential for future vaccine candidates.
2008-01-09T10:03:02Z
2008-01-09T10:03:02Z
2006-05-01
Article
Vaccines against typhoid fever. 2006, 24 (18):3804-11 Vaccine
0264-410X
16278037
10.1016/j.vaccine.2005.07.111
http://hdl.handle.net/10033/15855
Vaccine
en
oai:repository.helmholtz-hzi.de:10033/159132019-08-30T11:34:48Zcom_10033_620652col_10033_620666
2008-01-10T09:50:59Z
urn:hdl:10033/15913
An SopB-mediated immune escape mechanism of Salmonella enterica can be subverted to optimize the performance of live attenuated vaccine carrier strains.
Link, Claudia
Ebensen, Thomas
Ständner, Lothar
Déjosez, Marion
Reinhard, Elena
Rharbaoui, Faiza
Guzmán, Carlos A
Department of Vaccinology, Division of Microbiology, GBF-German Research Centre for Biotechnology, Mascheroder Weg 1, D-38124 Braunschweig, Germany.
Salmonellae have evolved several mechanisms to evade host clearance. Here, we describe the influence on bacterial immune escape of the effector protein SopB, which is translocated into the cytosol through a type III secretion system. Wild-type bacteria, as well as the sseC and aroA attenuated mutants exerted a stronger cytotoxic effect on dendritic cells (DC) than their SopB-deficient derivatives. Cells infected with the double sseC sopB, phoP sopB and aroA sopB mutants also exhibited higher expression of MHC, CD80, CD86 and CD54 molecules, and showed a stronger capacity to process and present an I-E(d)-restricted epitope from the influenza hemagglutinin (HA) to CD4+ cells from TCR-HA transgenic mice in vitro. The incorporation of an additional mutation into the sopB locus of the attenuated sseC, phoP and aroA mutants resulted in the stimulation of improved humoral and cellular immune responses following oral vaccination. The obtained results define a new potential immune escape strategy of this important pathogen, and also demonstrate that this mechanism can be subverted to optimize the immune responses elicited using Salmonella as a live vaccine carrier.
2008-01-10T09:50:59Z
2008-01-10T09:50:59Z
2006-07
Article
An SopB-mediated immune escape mechanism of Salmonella enterica can be subverted to optimize the performance of live attenuated vaccine carrier strains. 2006, 8 (8):2262-9 Microbes Infect.
1286-4579
16793312
10.1016/j.micinf.2006.04.013
http://hdl.handle.net/10033/15913
Microbes and infection / Institut Pasteur
en
oai:repository.helmholtz-hzi.de:10033/171922019-08-30T11:34:22Zcom_10033_620652col_10033_620666
2008-01-31T10:20:24Z
urn:hdl:10033/17192
Intranasal vaccination with recombinant outer membrane protein CD and adamantylamide dipeptide as the mucosal adjuvant enhances pulmonary clearance of Moraxella catarrhalis in an experimental murine model.
Becker, Pablo D
Bertot, Gustavo M
Souss, David
Ebensen, Thomas
Guzmán, Carlos A
Grinstein, Saúl
Virology Laboratory, Ricardo Gutiérrez Children's Hospital, Gallo 1330, 1425 Buenos Aires, Argentina.
Moraxella catarrhalis causes acute otitis media in children and lower respiratory tract infections in adults and elderly. In children the presence of antibodies against the highly conserved outer membrane protein CD correlates with protection against infection, suggesting that this protein may be useful as a vaccine antigen. However, native CD is difficult to purify, and it is still unclear if recombinant CD (rCD) is a valid alternative. We performed a side-by-side comparison of the immunogenicities and efficacies of vaccine formulations containing native CD and rCD with adamantylamide dipeptide as the mucosal adjuvant. Intranasal vaccination of mice stimulated the production of high CD-specific antibody titers in sera and of secretory immunoglobulin A in mucosal lavages, which cross-recognized both antigens. While vaccination with native CD increased the number of interleukin-2 (IL-2)- and gamma interferon-producing cells, rCD mainly stimulated IL-4-secreting cells. Nevertheless, efficient bacterial clearance was observed in the lungs of challenged mice receiving native CD and in the lungs of challenged mice receiving rCD (96% and 99%, respectively). Thus, rCD is a promising candidate for incorporation in vaccine formulations for use against M. catarrhalis.
2008-01-31T10:20:24Z
2008-01-31T10:20:24Z
2007-04
Article
Intranasal vaccination with recombinant outer membrane protein CD and adamantylamide dipeptide as the mucosal adjuvant enhances pulmonary clearance of Moraxella catarrhalis in an experimental murine model. 2007, 75 (4):1778-84 Infect. Immun.
0019-9567
17101651
10.1128/IAI.01081-06
http://hdl.handle.net/10033/17192
Infection and immunity
en
oai:repository.helmholtz-hzi.de:10033/189402019-08-30T11:37:00Zcom_10033_620652col_10033_620666
2008-02-22T09:37:33Z
urn:hdl:10033/18940
The bacterial second messenger cdiGMP exhibits promising activity as a mucosal adjuvant.
Ebensen, Thomas
Schulze, Kai
Riese, Peggy
Morr, Michael
Guzmán, Carlos A
Department of Vaccinology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, D-38124 Braunschweig, Germany.
The development of mucosal adjuvants is still a critical need in vaccinology. In the present work, we show that bis(3',5')-cyclic dimeric GMP (cdiGMP), a second messenger that modulates cell surface properties of several microorganisms, exerts potent activity as a mucosal adjuvant. BALB/c mice were immunized intranasally with the model antigen beta-galactosidase (beta-Gal) coadministered with cdiGMP. Animals receiving cdiGMP as an adjuvant showed significantly higher anti-beta-Gal immunoglobulin G (IgG) titers in sera than controls (i.e., 512-fold [P < 0.05]). Coadministration of cdiGMP also stimulated efficient beta-Gal-specific secretory IgA production in the lung (P < 0.016) and vagina (P < 0.036). Cellular immune responses were observed in response to both the beta-Gal protein and a peptide encompassing its major histocompatibility complex class I-restricted epitope. The IgG1-to-IgG2a ratio of anti-beta-Gal antibodies and the observed profiles of secreted cytokines suggest that a dominant Th1 response pattern is promoted by mucosal coadministration of cdiGMP. Finally, the use of cdiGMP as a mucosal adjuvant also led to the stimulation of in vivo cytotoxic T-lymphocyte responses in C57BL/6 mice intranasally immunized with ovalbumin and cdiGMP (up to 30% of specific lysis). The results obtained indicate that cdiGMP is a promising tool for the development of mucosal vaccines.
2008-02-22T09:37:33Z
2008-02-22T09:37:33Z
2007-08
Article
The bacterial second messenger cdiGMP exhibits promising activity as a mucosal adjuvant. 2007, 14 (8):952-8 Clin. Vaccine Immunol.
1556-6811
17567766
10.1128/CVI.00119-07
http://hdl.handle.net/10033/18940
Clinical and vaccine immunology : CVI
en
oai:repository.helmholtz-hzi.de:10033/197542019-08-30T11:30:58Zcom_10033_620652col_10033_620666
2008-03-05T09:45:09Z
urn:hdl:10033/19754
Replication-deficient mutant Herpes Simplex Virus-1 targets professional antigen presenting cells and induces efficient CD4+ T helper responses.
Fiorentini, Simona
Marconi, Peggy
Avolio, Manuela
Marini, Elena
Garrafa, Emirena
Caracciolo, Sonia
Rossi, Daniele
Bozac, Alexandra
Becker, Pablo D
Gentili, Francesca
Facchetti, Fabio
Guzman, Carlos A
Manservigi, Roberto
Caruso, Arnaldo
Department of Experimental and Applied Medicine, Section of Microbiology, University of Brescia Medical School, Piazzale Spedali Civili, 1, I-25123 Brescia, Italy.
Both neutralizing antibodies and cytotoxic T-cells are necessary to control a viral infection. However, vigorous T helper responses are essential for their elicitation and maintenance. Here we show that a recombinant replication-deficient Herpes Simplex Virus (HSV)-1 vector encoding the Human Immunodeficiency Virus (HIV)-1 matrix protein p17 (T0-p17) was capable of infecting professional antigen presenting cells (APCs) in vitro and in vivo. The injection of T0-p17 in the mouse dermis generated a strong p17-specific CD4+ T helper response preceding both p17-specific humoral and effector T cell responses. Moreover, we show that T0-p17 infection did not interfere with the endogenous processing of the transgene encoded antigen, since infected APCs were able to evoke a strong recall response in vitro. Our results demonstrate that replication-deficient HSV vectors can be appealing candidates for the development of vaccines able to trigger T helper responses.
2008-03-05T09:45:09Z
2008-03-05T09:45:09Z
2007-07
Article
Replication-deficient mutant Herpes Simplex Virus-1 targets professional antigen presenting cells and induces efficient CD4+ T helper responses. 2007, 9 (8):988-96 Microbes Infect.
1286-4579
17553721
10.1016/j.micinf.2007.04.001
http://hdl.handle.net/10033/19754
Microbes and infection / Institut Pasteur
en
oai:repository.helmholtz-hzi.de:10033/223322019-08-30T11:31:23Zcom_10033_620652col_10033_620666
2008-04-04T12:35:48Z
urn:hdl:10033/22332
Modulation of chemokine gene expression by Shiga-toxin producing Escherichia coli belonging to various origins and serotypes.
Gobert, Alain P
Coste, Alix
Guzman, Carlos A
Vareille, Marjolaine
Hindré, Thomas
de Sablet, Thibaut
Girardeau, Jean-Pierre
Martin, Christine
Institut National de la Recherche Agronomique (INRA), UR454 Unité de Microbiologie, Centre de Theix, 63122 Saint-Genès-Champanelle, France.
Infection with Shiga-toxin producing Escherichia coli (STEC) may result in the development of the haemolytic-uremic syndrome (HUS), the main cause of acute renal failure in children. While O157:H7 STEC are associated with large outbreaks of HUS, it is difficult to predict whether a non-O157:H7 isolate can be pathogenic for humans. The mucosal innate immune response plays a central role in the pathogenesis of HUS; therefore, we compared the induction of IL-8 and CCL20 in human colon epithelial cells infected with strains belonging to different serotypes, isolated from cattle or from HUS patients. No correlation was observed between strain virulence and chemokine gene expression. Rather, the genetic background of the strains seems to determine the chemokine gene expression profile. Investigating the contribution of different bacterial factors in this process, we show that the type III secretion system of O157:H7 bacteria, but not the intimate adhesion, is required to stimulate the cells. In addition, H7, H10, and H21 flagellins are potent inducers of chemokine gene expression when synthesized in large amount.
2008-04-04T12:35:48Z
2008-04-04T12:35:48Z
2008-02
Article
Modulation of chemokine gene expression by Shiga-toxin producing Escherichia coli belonging to various origins and serotypes. 2008, 10 (2):159-65 Microbes Infect.
1286-4579
18248761
10.1016/j.micinf.2007.10.018
http://hdl.handle.net/10033/22332
Microbes and infection / Institut Pasteur
en
oai:repository.helmholtz-hzi.de:10033/230722019-08-30T11:37:44Zcom_10033_620652col_10033_620666
2008-04-11T12:43:20Z
urn:hdl:10033/23072
Oral vaccination with Salmonella enterica as a cruzipain-DNA delivery system confers protective immunity against Trypanosoma cruzi.
Cazorla, Silvia I
Becker, Pablo D
Frank, Fernanda M
Ebensen, Thomas
Sartori, María J
Corral, Ricardo S
Malchiodi, Emilio L
Guzmán, Carlos A
Instituto de Estudios de la Inmunidad Humoral, CONICET-UBA, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
To stimulate both local and systemic immune responses against Trypanosoma cruzi, Salmonella enterica serovar Typhimurium aroA was exploited as a DNA delivery system for cruzipain (SCz). In a murine model we compared SCz alone (GI) or coadministered with Salmonella carrying a plasmid encoding granulocyte-macrophage colony-stimulating factor (GII), as well as protocols in which SCz priming was followed by boosting with recombinant cruzipain (rCz) admixed with either CpG-ODN (GIII) or MALP-2, a synthetic derivative of a macrophage-activating lipopeptide of 2 kDa from Mycoplasma fermentans (GIV). The results showed that protocols that included four oral doses of SCz (GI) elicited mainly a mucosal response characterized by immunoglobulin A (IgA) secretion and proliferation of gut-associated lymphoid tissue cells, with weak systemic responses. In contrast, the protocol that included a boost with rCz plus CpG (GIII) triggered stronger systemic responses in terms of Cz-specific serum IgG titers, splenocyte proliferation, gamma interferon (IFN-gamma) secretion, and delayed-type hypersensitivity response. Trypomastigote challenge of vaccinated mice resulted in significantly lower levels of parasitemia compared to controls. Protection was abolished by depletion of either CD4+ or CD8+ T cells. Parasite control was also evident from the reduction of tissue damage, as revealed by histopathologic studies and serum levels of enzymes that are markers of muscle injury in chronic Chagas' disease (i.e., creatine kinase, aspartate aminotransferase, and lactate dehydrogenase). Enhanced release of IFN-gamma and interleukin-2 was observed in GI and GII upon restimulation of splenocytes in the nonparasitic phase of infection. Our results indicate that Salmonella-mediated delivery of Cz-DNA by itself promotes the elicitation of an immune response that controls T. cruzi infection, thereby reducing parasite loads and subsequent damage to muscle tissues.
2008-04-11T12:43:20Z
2008-04-11T12:43:20Z
2008-01
Article
Oral vaccination with Salmonella enterica as a cruzipain-DNA delivery system confers protective immunity against Trypanosoma cruzi. 2008, 76 (1):324-33 Infect. Immun.
1098-5522
17967857
10.1128/IAI.01163-07
http://hdl.handle.net/10033/23072
Infection and immunity
en
oai:repository.helmholtz-hzi.de:10033/238522019-08-30T11:34:22Zcom_10033_620652col_10033_620666
2008-04-21T08:57:33Z
urn:hdl:10033/23852
In vivo gene regulation in Salmonella spp. by a salicylate-dependent control circuit.
Royo, José Luis
Becker, Pablo Daniel
Camacho, Eva María
Cebolla, Angel
Link, Claudia
Santero, Eduardo
Guzmán, Carlos Alberto
Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide-Consejo Superior de Investigaciones Científicas, Carretera, Utrera, Km 1, E-41013 Sevilla, Spain.
Systems allowing tightly regulated expression of prokaryotic genes in vivo are important for performing functional studies of bacterial genes in host-pathogen interactions and establishing bacteria-based therapies. We integrated a regulatory control circuit activated by acetyl salicylic acid (ASA) in attenuated Salmonella enterica that carries an expression module with a gene of interest under control of the XylS2-dependent Pm promoter. This resulted in 20-150-fold induction ex vivo. The regulatory circuit was also efficiently induced by ASA when the bacteria resided in eukaryotic cells, both in vitro and in vivo. To validate the circuit, we administered Salmonella spp., carrying an expression module encoding the 5-fluorocytosine-converting enzyme cytosine deaminase in the bacterial chromosome or in a plasmid, to mice with tumors. Induction with ASA before 5-fluorocytosine administration resulted in a significant reduction of tumor growth. These results demonstrate the usefulness of the regulatory control circuit to selectively switch on gene expression during bacterial infection.
2008-04-21T08:57:33Z
2008-04-21T08:57:33Z
2007-11
Article
In vivo gene regulation in Salmonella spp. by a salicylate-dependent control circuit. 2007, 4 (11):937-42 Nat. Methods
1548-7091
17922017
10.1038/nmeth1107
http://hdl.handle.net/10033/23852
Nature methods
en
oai:repository.helmholtz-hzi.de:10033/250802019-08-30T11:34:48Zcom_10033_620652col_10033_620666
2008-05-08T10:46:42Z
urn:hdl:10033/25080
Prime-boost immunization with cruzipain co-administered with MALP-2 triggers a protective immune response able to decrease parasite burden and tissue injury in an experimental Trypanosoma cruzi infection model.
Cazorla, SI
Frank, FM
Becker, PD
Corral, RS
Guzmán, CA
Malchiodi, EL
Cátedra de Inmunología and Instituto de Estudios de la Inmunidad Humoral (IDEHU), CONICET-UBA, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junin 956 4to P, 1113 Buenos Aires, Argentina; Departamento de Microbiología, Parasitología e Inmunología, Facultad de Medicina, Universidad de Buenos Aires, Argentina; Department of Vaccinology, Helmholtz Centre for Infection Research, Inhoffenstraße 7, D-38124 Braunschweig, Germany.
Cruzipain (Cz), a key Trypanosoma cruzi enzyme, is a main candidate antigen for vaccines against Chagas' disease. We evaluated a vaccination protocol based on intradermal priming with recombinant Cz and intranasal boosting with rCz co-administered with a derivative of the TLR2/6 agonist MALP-2. Vaccination triggered strong systemic and mucosal antibody responses, and a vigorous cell-mediated immunity characterized by lymphoproliferation, DTH reactivity and IFN-gamma production. The immune responses protected against a lethal trypomastigote challenge and, upon sub-lethal infection, immunized mice showed reduction of tissue damage and normal enzymatic markers of muscle injury. This prime-boost regimen appears promising for further development, since warranted survival, provided efficient control of parasite load and restricted inflammatory myopathy.
2008-05-08T10:46:42Z
2008-05-08T10:46:42Z
2008-04-07
Article
Prime-boost immunization with cruzipain co-administered with MALP-2 triggers a protective immune response able to decrease parasite burden and tissue injury in an experimental Trypanosoma cruzi infection model. 2008, 26 (16):1999-2009 Vaccine
0264-410X
18342408
10.1016/j.vaccine.2008.02.011
http://hdl.handle.net/10033/25080
Vaccine
oai:repository.helmholtz-hzi.de:10033/303932019-08-30T11:35:13Zcom_10033_620652col_10033_620666
2008-06-24T11:32:05Z
urn:hdl:10033/30393
Intramammary application of non-methylated-CpG oligodeoxynucleotides (CpG) inhibits both local and systemic mammary carcinogenesis in female BALB/c Her-2/neu transgenic mice.
Mastini, Cristina
Becker, Pablo D
Iezzi, Manuela
Curcio, Claudia
Musiani, Piero
Forni, Guido
Cavallo, Federica
Guzmán, Carlos A
Molecular Biotechnology Center, Department of Clinical and Biological Sciences, University of Torino, 10126 Torino, Italy.
CpG are powerful drugs activating the innate immune system. In this study, the ability of their intramammary administration in impeding the devastating progression of carcinogenesis in all the mammary glands of female BALB/c mice transgenic for the rat neu transforming oncogene was assessed. Starting when in situ carcinomas were scattered over all their mammary glands (week 10), mice received CpG injections in the stroma of the fourth left gland. Local neoplastic progression was inhibited by six monthly administrations. CpG not only delayed the onset of carcinomas in the injected gland, but also hampered their progression. Extended latency was observed for tumors in glands both close to and far from the injection site. When the experiment ended (week 45), no tumors were palpable in 67% of the injected glands and a markedly impaired tumor growth was evident in the others. An impressive local infiltrate of CD11b(+) cells with the morphologic features of macrophages, plasma cells, B220(+) B cells, and CD4(+) and CD8(+) T cells was quickly recruited to the CpG-treated glands. High quantities of IFN-gamma producing cells were only present in the ipsilateral axillary draining lymph nodes of the treated glands. Enhanced natural killer (NK) lytic activity was also detected in the spleens. Inhibition of progression was weaker when only four injections were given, and abolished by in vivo depletion of NK cells. CpG monotherapy is thus effective in an aggressive model of autochthonous cancer. The results strongly support the administration of CpG as a local monotherapy of multiple invasive microscopic lesions.
2008-06-24T11:32:05Z
2008-06-24T11:32:05Z
2008-05
Article
Intramammary application of non-methylated-CpG oligodeoxynucleotides (CpG) inhibits both local and systemic mammary carcinogenesis in female BALB/c Her-2/neu transgenic mice. 2008, 8 (3):230-42notCurr Cancer Drug Targets
1873-5576
18473736
http://hdl.handle.net/10033/30393
Current cancer drug targets
en
oai:repository.helmholtz-hzi.de:10033/422542019-08-30T11:36:05Zcom_10033_620652col_10033_620666
2008-12-11T14:58:24Z
urn:hdl:10033/42254
The missing link: Bordetella petrii is endowed with both the metabolic versatility of environmental bacteria and virulence traits of pathogenic Bordetellae.
Gross, Roy
Guzman, Carlos A
Sebaihia, Mohammed
dos Santos, Vítor A P Martins
Pieper, Dietmar H
Koebnik, Ralf
Lechner, Melanie
Bartels, Daniela
Buhrmester, Jens
Choudhuri, Jomuna V
Ebensen, Thomas
Gaigalat, Lars
Herrmann, Stefanie
Khachane, Amit N
Larisch, Christof
Link, Stefanie
Linke, Burkhard
Meyer, Folker
Mormann, Sascha
Nakunst, Diana
Rückert, Christian
Schneiker-Bekel, Susanne
Schulze, Kai
Vorhölter, Frank-Jörg
Yevsa, Tetyana
Engle, Jacquelyn T
Goldman, William E
Pühler, Alfred
Göbel, Ulf B
Goesmann, Alexander
Blöcker, Helmut
Kaiser, Olaf
Martinez-Arias, Rosa
Chair of Microbiology, Biocenter, University of Würzburg, Am Hubland, D-97074 Würzburg, Germany. roy@biozentrum.uni-wuerzburg.de
BACKGROUND: Bordetella petrii is the only environmental species hitherto found among the otherwise host-restricted and pathogenic members of the genus Bordetella. Phylogenetically, it connects the pathogenic Bordetellae and environmental bacteria of the genera Achromobacter and Alcaligenes, which are opportunistic pathogens. B. petrii strains have been isolated from very different environmental niches, including river sediment, polluted soil, marine sponges and a grass root. Recently, clinical isolates associated with bone degenerative disease or cystic fibrosis have also been described. RESULTS: In this manuscript we present the results of the analysis of the completely annotated genome sequence of the B. petrii strain DSMZ12804. B. petrii has a mosaic genome of 5,287,950 bp harboring numerous mobile genetic elements, including seven large genomic islands. Four of them are highly related to the clc element of Pseudomonas knackmussii B13, which encodes genes involved in the degradation of aromatics. Though being an environmental isolate, the sequenced B. petrii strain also encodes proteins related to virulence factors of the pathogenic Bordetellae, including the filamentous hemagglutinin, which is a major colonization factor of B. pertussis, and the master virulence regulator BvgAS. However, it lacks all known toxins of the pathogenic Bordetellae. CONCLUSION: The genomic analysis suggests that B. petrii represents an evolutionary link between free-living environmental bacteria and the host-restricted obligate pathogenic Bordetellae. Its remarkable metabolic versatility may enable B. petrii to thrive in very different ecological niches.
2008-12-11T14:58:24Z
2008-12-11T14:58:24Z
2008
Article
The missing link: Bordetella petrii is endowed with both the metabolic versatility of environmental bacteria and virulence traits of pathogenic Bordetellae. 2008, 9:449 BMC Genomics
1471-2164
18826580
10.1186/1471-2164-9-449
http://hdl.handle.net/10033/42254
BMC genomics
en
http://www.biomedcentral.com/1471-2164/9/449
oai:repository.helmholtz-hzi.de:10033/483532019-08-30T11:36:04Zcom_10033_620652col_10033_620666
2009-02-03T09:09:46Z
urn:hdl:10033/48353
Lactobacillus reuteri DSM 20016 produces cobalamin-dependent diol dehydratase in metabolosomes and metabolizes 1,2-propanediol by disproportionation.
Sriramulu, Dinesh Diraviam
Liang, Mingzhi
Hernandez-Romero, Diana
Raux-Deery, Evelyne
Lünsdorf, Heinrich
Parsons, Joshua B
Warren, Martin J
Prentice, Michael B
Department of Microbiology, University College Cork, Cork, Ireland.
A Lactobacillus reuteri strain isolated from sourdough is known to produce the vitamin cobalamin. The organism requires this for glycerol cofermentation by a cobalamin-dependent enzyme, usually termed glycerol dehydratase, in the synthesis of the antimicrobial substance reuterin. We show that the cobalamin-synthesizing capacity of another L. reuteri strain (20016, the type strain, isolated from the human gut and recently sequenced as F275) is genetically and phenotypically linked, as in the Enterobacteriaceae, to the production of a cobalamin-dependent enzyme which is associated with a bacterial microcompartment (metabolosome) and known as diol dehydratase. We show that this enzyme allows L. reuteri to carry out a disproportionation reaction converting 1,2-propanediol to propionate and propanol. The wide distribution of this operon suggests that it is adapted to horizontal transmission between bacteria. However, there are significant genetic and phenotypic differences between the Lactobacillus background and the Enterobacteriaceae. Electron microscopy reveals that the bacterial microcompartment in L. reuteri occupies a smaller percentage of the cytoplasm than in gram-negative bacteria. DNA sequence data show evidence of a regulatory control mechanism different from that in gram-negative bacteria, with the presence of a catabolite-responsive element (CRE) sequence immediately upstream of the pdu operon encoding diol dehydratase and metabolosome structural genes in L. reuteri. The metabolosome-associated diol dehydratase we describe is the only candidate glycerol dehydratase present on inspection of the L. reuteri F275 genome sequence.
2009-02-03T09:09:46Z
2009-02-03T09:09:46Z
2008-07
Article
Lactobacillus reuteri DSM 20016 produces cobalamin-dependent diol dehydratase in metabolosomes and metabolizes 1,2-propanediol by disproportionation. 2008, 190 (13):4559-67 J. Bacteriol.
1098-5530
18469107
10.1128/JB.01535-07
http://hdl.handle.net/10033/48353
Journal of bacteriology
en
oai:repository.helmholtz-hzi.de:10033/521932019-08-30T11:36:05Zcom_10033_620652col_10033_620666
2009-03-05T09:01:48Z
urn:hdl:10033/52193
Structural and functional characterization of human Iba proteins.
Schulze, Jörg O
Quedenau, Claudia
Roske, Yvette
Adam, Thomas
Schüler, Herwig
Behlke, Joachim
Turnbull, Andrew P
Sievert, Volker
Scheich, Christoph
Mueller, Uwe
Heinemann, Udo
Büssow, Konrad
Max Delbrück Center for Molecular Medicine, Berlin, Germany.
Iba2 is a homolog of ionized calcium-binding adapter molecule 1 (Iba1), a 17-kDa protein that binds and cross-links filamentous actin (F-actin) and localizes to membrane ruffles and phagocytic cups. Here, we present the crystal structure of human Iba2 and its homodimerization properties, F-actin cross-linking activity, cellular localization and recruitment upon bacterial invasion in comparison with Iba1. The Iba2 structure comprises two central EF-hand motifs lacking bound Ca2+. Iba2 crystallized as a homodimer stabilized by a disulfide bridge and zinc ions. Analytical ultracentrifugation revealed a different mode of dimerization under reducing conditions that was independent of Ca2+. Furthermore, no binding of Ca2+ up to 0.1 mM was detected by equilibrium dialysis. Correspondingly, Iba EF-hand motifs lack residues essential for strong Ca2+ coordination. Sedimentation experiments and microscopy detected pronounced, indistinguishable F-actin binding and cross-linking activity of Iba1 and Iba2 with induction of F-actin bundles. Fluorescent Iba fusion proteins were expressed in HeLa cells and co-localized with F-actin. Iba1 was recruited into cellular projections to a larger extent than Iba2. Additionally, we studied Iba recruitment in a Shigella invasion model that induces cytoskeletal rearrangements. Both proteins were recruited into the bacterial invasion zone and Iba1 was again concentrated slightly higher in the cellular extensions.
2009-03-05T09:01:48Z
2009-03-05T09:01:48Z
2008-09
Article
Structural and functional characterization of human Iba proteins. 2008, 275 (18):4627-40 FEBS J.
1742-464X
18699778
10.1111/j.1742-4658.2008.06605.x
http://hdl.handle.net/10033/52193
The FEBS journal
en
oai:repository.helmholtz-hzi.de:10033/522532019-08-30T11:36:05Zcom_10033_620652col_10033_620666
2009-03-05T09:31:56Z
urn:hdl:10033/52253
PspA can form large scaffolds in Escherichia coli.
Standar, Kerstin
Mehner, Denise
Osadnik, Hendrik
Berthelmann, Felix
Hause, Gerd
Lünsdorf, Heinrich
Brüser, Thomas
Institute of Biology/Microbiology, University of Halle-Wittenberg, Kurt-Mothes-Strasse 3, D-06120 Halle, Germany.
The phage shock protein A (PspA) of Escherichia coli stabilizes the cytoplasmic membrane under stress conditions. Here we demonstrate that PspA can form hollow spherical or prolate spheroidal particles of about 30-40nm diameter with a scaffold-like arrangement of protein subunits at the surface. The 'PspA-scaffold' is the basic structure that is common to all particles. The PspA-scaffold may be of fundamental importance, as it could allow PspA to stabilize the integrity of membranes through numerous contact points over a large surface area.
2009-03-05T09:31:56Z
2009-03-05T09:31:56Z
2008-10-29
Article
PspA can form large scaffolds in Escherichia coli. 2008, 582 (25-26):3585-9 FEBS Lett.
0014-5793
18789328
10.1016/j.febslet.2008.09.002
http://hdl.handle.net/10033/52253
FEBS letters
en
oai:repository.helmholtz-hzi.de:10033/712132019-08-30T11:35:39Zcom_10033_620652col_10033_620666
2009-06-23T08:07:33Z
urn:hdl:10033/71213
Evolution of hepatitis C virus hypervariable region 1 in immunocompetent children born to HCV-infected mothers.
Gismondi, M I
Becker, P D
Díaz Carrasco, J M
Guzmán, C A
Campos, R H
Preciado, M V
Laboratorio de Biología Molecular, División Patología, Hospital de Niños Ricardo Gutiérrez, Buenos Aires, Argentina.
Hepatitis C virus (HCV) hypervariable region 1 (HVR1) is the most variable region of the viral genome and its heterogeneity reflects the virus-host interplay during chronicity. Paediatric HCV-infected patients develop liver disease with typical clinical features. Here, the evolution of HVR1 and its adjacent regions were ascertained in plasma samples of two HCV-positive children during a 5-year follow-up period. We report an almost complete conservation of the HVR1 amino acid sequence over time, with underlying nucleotide variability both within and outside HVR1, suggesting some kind of constraint on virus evolution, particularly within HVR1. Although overall d(N)/d(S) rates [rates of nonsynonymous nucleotide substitutions per nonsynonymous site (d(N)) and synonymous nucleotide substitutions per synonymous site (d(S))] were <1 in both patients, a high resolution analysis of selection pressures exerted at the codon level revealed few sites subject to selection and an absolute predominance of invariable positions within HVR1. The HVR1 amino acid sequences showed the antigenic properties expected for this region. Taken together, these data suggest peculiar evolutionary dynamics in our patients, which could be attributed to a mechanism of nucleotide invariability along with purifying selection operating on the HVR1. The lack of HVR1 variability may reflect the adaptation of the virus to a particular environment within each patient or a phenomenon of immune tolerance generated in these immunocompetent patients earlier in life.
2009-06-23T08:07:33Z
2009-06-23T08:07:33Z
2009-05
Article
Evolution of hepatitis C virus hypervariable region 1 in immunocompetent children born to HCV-infected mothers. 2009, 16 (5):332-9 J. Viral Hepat.
1365-2893
19228286
10.1111/j.1365-2893.2009.01071.x
http://hdl.handle.net/10033/71213
Journal of viral hepatitis
en
oai:repository.helmholtz-hzi.de:10033/874132019-08-30T11:34:48Zcom_10033_620652col_10033_620666
2009-12-04T12:59:14Z
urn:hdl:10033/87413
A prime-boost vaccination protocol optimizes immune responses against the nucleocapsid protein of the SARS coronavirus.
Schulze, Kai
Staib, Caroline
Schätzl, Hermann M
Ebensen, Thomas
Erfle, Volker
Guzman, Carlos A
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, D-38124 Braunschweig, Germany.
Severe acute respiratory syndrome (SARS) is a serious infectious disease caused by the SARS coronavirus. We assessed the potential of prime-boost vaccination protocols based on the nucleocapsid (NC) protein co-administered with a derivative of the mucosal adjuvant MALP-2 or expressed by modified Vaccinia virus Ankara (MVA-NC) to stimulate humoral and cellular immune responses at systemic and mucosal levels. The obtained results demonstrated that strong immune responses can be elicited both at systemic and mucosal levels following a heterologous prime-boost vaccination protocol consisting in priming with NC protein add-mixed with MALP-2 by intranasal route and boosting with MVA-NC by intramuscular route.
2009-12-04T12:59:14Z
2009-12-04T12:59:14Z
2008-12-02
Article
A prime-boost vaccination protocol optimizes immune responses against the nucleocapsid protein of the SARS coronavirus. 2008, 26 (51):6678-84 Vaccine
0264-410X
18805454
10.1016/j.vaccine.2008.09.006
http://hdl.handle.net/10033/87413
Vaccine
en
oai:repository.helmholtz-hzi.de:10033/1163242019-08-30T11:35:39Zcom_10033_620652col_10033_620666
2010-11-25T15:41:46Z
urn:hdl:10033/116324
Modified vaccinia virus Ankara exerts potent immune modulatory activities in a murine model.
Nörder, Miriam
Becker, Pablo D
Drexler, Ingo
Link, Claudia
Erfle, Volker
Guzmán, Carlos A
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
BACKGROUND: Modified vaccinia virus Ankara (MVA), a highly attenuated strain of vaccinia virus, has been used as vaccine delivery vector in preclinical and clinical studies against infectious diseases and malignancies. Here, we investigated whether an MVA which does not encode any antigen (Ag) could be exploited as adjuvant per se. METHODOLOGY/PRINCIPAL FINDINGS: We showed that dendritic cells infected in vitro with non-recombinant (nr) MVA expressed maturation and activation markers and were able to efficiently present exogenously pulsed Ag to T cells. In contrast to the dominant T helper (Th) 1 biased responses elicited against Ags produced by recombinant MVA vectors, the use of nrMVA as adjuvant for the co-administered soluble Ags resulted in a long lasting mixed Th1/Th2 responses. CONCLUSIONS/SIGNIFICANCE: These findings open new ways to potentiate and modulate the immune responses to vaccine Ags depending on whether they are co-administered with MVA or encoded by recombinant viruses.
2010-11-25T15:41:46Z
2010-11-25T15:41:46Z
2010
Article
Modified vaccinia virus Ankara exerts potent immune modulatory activities in a murine model. 2010, 5 (6):e11400 PLoS ONE
1932-6203
20628596
10.1371/journal.pone.0011400
http://hdl.handle.net/10033/116324
PloS one
en
oai:repository.helmholtz-hzi.de:10033/1193872019-08-30T11:37:00Zcom_10033_620652col_10033_620666
2011-01-14T11:31:08Z
urn:hdl:10033/119387
Redirection of the immune response to the functional catalytic domain of the cystein proteinase cruzipain improves protective immunity against Trypanosoma cruzi infection.
Cazorla, Silvia I
Frank, Fernanda M
Becker, Pablo D
Arnaiz, María
Mirkin, Gerardo A
Corral, Ricardo S
Guzmán, Carlos A
Malchiodi, Emilio L
Cátedra de Inmunología and Instituto de Estudios de la Inmunidad Humoral (IDEHU), Universidad de BuenosAires, 1113 Buenos Aires, Argentina.
Despite the strong immune responses elicited after natural infection with Trypanosoma cruzi or vaccination against it, parasite survival suggests that these responses are insufficient or inherently inadequate. T. cruzi contains a major cystein proteinase, cruzipain, which has a catalytic N-terminal domain and a C-terminal extension. Immunizations that employed recombinant cruzipain or its N- and C-terminal domains allowed evaluation of the ability of cruzipain to circumvent responses against the catalytic domain. This phenomenon is not a property of the parasite but of cruzipain itself, because recombinant cruzipain triggers a response similar to that of cruzipain during natural or experimental infection. Cruzipain is not the only antigen with a highly immunogenic region of unknown function that somehow protects an essential domain for parasite survival. However, our studies show that this can be reverted by using the N-terminal domain as a tailored immunogen able to redirect host responses to provide enhanced protection.
2011-01-14T11:31:08Z
2011-01-14T11:31:08Z
2010-07-01
Article
Redirection of the immune response to the functional catalytic domain of the cystein proteinase cruzipain improves protective immunity against Trypanosoma cruzi infection. 2010, 202 (1):136-44 J. Infect. Dis.
1537-6613
20497050
10.1086/652872
http://hdl.handle.net/10033/119387
The Journal of infectious diseases
en
oai:repository.helmholtz-hzi.de:10033/1233332019-08-30T11:36:59Zcom_10033_620652col_10033_620666
2011-03-02T12:55:22Z
urn:hdl:10033/123333
Immune modulators with defined molecular targets: cornerstone to optimize rational vaccine design.
Ebensen, Thomas
Guzmán, Carlos A
Department of Vaccinology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
Vaccination remains the most valuable tool for preventing infectious diseases. However, the performance of many existing vaccines should be improved and there are diseases for which vaccines are still not available. The use of well-defined antigens for the generation of subunit vaccines has led to products with an improved safety profile. However, purified antigens are usually poorly immunogenic, making essential the use of adjuvants. Despite the fact that adjuvants have been used to increase the immunogenicity of vaccines for more than 70 years, only a handful has been licensed for human use (e.g., aluminium salts, the micro-fluidized squalene-in-water emulsion MF59 and monophosphoryl lipid A). Thus, the development of new adjuvants which are able to promote broad and sustained immune responses at systemic and mucosal levels still remains as a major challenge in vaccinology. Recent advances in our understanding of the immune system have facilitated the identification of new biological targets for screening programs aimed at the discovery of novel immune stimulators. This resulted in the identification of new candidate adjuvants, which made possible the modulation of the immune responses elicited according to specific needs. A number of promising adjuvants which are currently under preclinical or clinical development will be described in this review.
2011-03-02T12:55:22Z
2011-03-02T12:55:22Z
2011-03-02T12:55:22Z
Article
Immune modulators with defined molecular targets: cornerstone to optimize rational vaccine design., 4 (1):13-22 Hum Vaccin
1554-8619
18376145
http://hdl.handle.net/10033/123333
Human vaccines
en
oai:repository.helmholtz-hzi.de:10033/1238822019-08-30T11:37:23Zcom_10033_620652col_10033_620666
2011-03-08T15:08:42Z
urn:hdl:10033/123882
High-resolution in situ genotyping of Legionella pneumophila populations in drinking water by multiple-locus variable-number tandem-repeat analysis using environmental DNA.
Kahlisch, Leila
Henne, Karsten
Draheim, Josefin
Brettar, Ingrid
Höfle, Manfred G
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research (HZI), 38124 Braunschweig, Germany.
Central to the understanding of infections by the waterborne pathogen Legionella pneumophila is its detection at the clonal level. Currently, multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) of L. pneumophila isolates can be used as a tool for high-resolution genotyping. Since L. pneumophila is difficult to isolate, the isolation of outbreak strains often fails due to a viable but nonculturable (VBNC) state of the respective environmental population. Therefore, we developed a cultivation-independent approach to detect single clones in drinking water. This approach is based on the extraction of DNA from drinking water followed by PCR using a set of eight VNTR primer pairs necessary for MLVA genotyping of L. pneumophila. The PCR amplicons were analyzed by single-strand conformation polymorphism (SSCP) and capillary electrophoresis to obtain the respective MLVA profiles. Parallel to the high-resolution analysis, we used the same environmental DNA to quantify the number of L. pneumophila cells in drinking water using real-time PCR with 16S rRNA gene-targeted primers. We used a set of drinking water samples from a small-scale drinking water network to test our approach. With these samples we demonstrated that the developed approach was directly applicable to DNA obtained from drinking water. We were able to detect more L. pneumophila MLVA genotypes in drinking water than we could detect by isolation. Our approach could be a valuable tool to identify outbreak strains even after the outbreak has occurred and has the potential to be applied directly to clinical material.
2011-03-08T15:08:42Z
2011-03-08T15:08:42Z
2010-09
Article
High-resolution in situ genotyping of Legionella pneumophila populations in drinking water by multiple-locus variable-number tandem-repeat analysis using environmental DNA. 2010, 76 (18):6186-95 Appl. Environ. Microbiol.
1098-5336
20656879
10.1128/AEM.00416-10
http://hdl.handle.net/10033/123882
Applied and environmental microbiology
en
oai:repository.helmholtz-hzi.de:10033/2039122019-08-30T11:37:00Zcom_10033_620652col_10033_620666
2012-01-20T14:33:28Z
urn:hdl:10033/203912
Generation of human antigen-specific monoclonal IgM antibodies using vaccinated "human immune system" mice.
Becker, Pablo D
Legrand, Nicolas
van Geelen, Caroline M M
Noerder, Miriam
Huntington, Nicholas D
Lim, Annick
Yasuda, Etsuko
Diehl, Sean A
Scheeren, Ferenc A
Ott, Michael
Weijer, Kees
Wedemeyer, Heiner
Di Santo, James P
Beaumont, Tim
Guzman, Carlos A
Spits, Hergen
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research (HZI), Braunschweig, Germany.
Passive transfer of antibodies not only provides immediate short-term protection against disease, but also can be exploited as a therapeutic tool. However, the 'humanization' of murine monoclonal antibodies (mAbs) is a time-consuming and expensive process that has the inherent drawback of potentially altering antigenic specificity and/or affinity. The immortalization of human B cells represents an alternative for obtaining human mAbs, but relies on the availability of biological samples from vaccinated individuals or convalescent patients. In this work we describe a novel approach to generate fully human mAbs by combining a humanized mouse model with a new B cell immortalization technique.
2012-01-20T14:33:28Z
2012-01-20T14:33:28Z
2010
Article
Generation of human antigen-specific monoclonal IgM antibodies using vaccinated "human immune system" mice. 2010, 5 (10) PLoS ONE
1932-6203
20957227
10.1371/journal.pone.0013137
http://hdl.handle.net/10033/203912
PloS one
en
oai:repository.helmholtz-hzi.de:10033/1346202019-08-30T11:29:47Zcom_10033_620652col_10033_620666
2011-06-27T14:21:00Z
urn:hdl:10033/134620
Porcine pulmonary angiotensin I-converting enzyme--biochemical characterization and spatial arrangement of the N- and C-domains by three-dimensional electron microscopic reconstruction.
Chen, Hui-Ling
Lünsdorf, Heinrich
Hecht, Hans-Jürgen
Tsai, Hsin
Development Center for Biotechnology, Taipei County 221, Taiwan, ROC.
The somatic angiotensin I-converting enzyme (sACE; peptidyl-dipeptidase A; EC 3.4.15.1) was isolated from pig lung and purified to homogeneity. The purified enzyme has a molecular mass of about 180 kDa. Upon proteolytic cleavage, two approximately 90 kDa fragments were obtained and identified by amino-terminal sequence analysis as the N- and C-domains of sACE. Both purified domains were shown to be catalytically active. A 2.3 nm resolution model of sACE was obtained by three-dimensional electron microscopic reconstruction of negatively stained sACE particles, based on atomic X-ray data fitting. Our model shows for the first time the relative orientation of the sACE catalytically active domains and their spatial distance.
2011-06-27T14:21:00Z
2011-06-27T14:21:00Z
2010-08
Article
Porcine pulmonary angiotensin I-converting enzyme--biochemical characterization and spatial arrangement of the N- and C-domains by three-dimensional electron microscopic reconstruction. 2010, 41 (6):674-85 Micron
1878-4291
20427191
10.1016/j.micron.2010.01.005
http://hdl.handle.net/10033/134620
Micron (Oxford, England : 1993)
en
oai:repository.helmholtz-hzi.de:10033/1350592019-08-30T11:25:11Zcom_10033_620652col_10033_620666
2011-07-01T13:53:14Z
urn:hdl:10033/135059
The NKT cell ligand αgalactosylceramide suppresses allergic airway inflammation by induction of a Th1 response.
Knothe, S
Mutschler, V
Rochlitzer, S
Winkler, C
Ebensen, T
Guzman, C A
Hohlfeld, J
Braun, A
Muller, M
Fraunhofer Institute for Toxicology and Experimental Medicine, Department of Immunology, Allergology and Immunotoxicology, Nikolai-Fuchs-Str. 1, 30625 Hannover, Germany.
One experimental approach for the treatment of allergic reactions is the stimulation of immunoregulatory NKT cells with the synthetic glycolipid αgalactosylceramide. For a first evaluation of the immunomodulatory potential of αGalCerMPEG a human in vitro allergy model was exploited. Acting as an adjuvant, the glycolipid induced an enhanced Th1-biased allergen-specific immune response of autologous lymphocytes. In a mouse model of allergic airway inflammation, αGalCerMPEG-activated NKT cells promoted a cytokine environment in the spleen, leading to priming of Th1 cells. The shift towards a Th1-dominated allergen-specific immune response thus might mediate the abrogation of allergic airway inflammation and thereby might provide a valid option for therapeutic intervention.
2011-07-01T13:53:14Z
2011-07-01T13:53:14Z
2011-06-06
Article
The NKT cell ligand αgalactosylceramide suppresses allergic airway inflammation by induction of a Th1 response. 2011, 29 (25):4249-55 Vaccine
1873-2518
21463684
10.1016/j.vaccine.2011.03.068
http://hdl.handle.net/10033/135059
Vaccine
en
oai:repository.helmholtz-hzi.de:10033/1355742019-08-30T11:30:58Zcom_10033_620652col_10033_620666
2011-07-07T14:19:36Z
urn:hdl:10033/135574
Local treatment with BPPcysMPEG reduces allergic airway inflammation in sensitized mice.
Knothe, S
Mutschler, V
Rochlitzer, S
Winkler, C
Ebensen, T
Guzman, C A
Hohlfeld, J
Braun, A
Muller, M
Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM), Department of Immunology, Allergology and Immunotoxicology, Hannover, Germany.
According to the hygiene hypothesis, triggering the immune system with microbial components during childhood balances the inherent Th2 bias. In contrast, specific immunotherapy involves exposure of the patient to the allergen in order to achieve desensitization to subsequent contact. In a human in vitro allergy model the potential of the TLR2/6 agonist BPPcysMPEG to modulate antigen presenting cells and allergen-specific immune responses was evaluated. Specific immunomodulation via co-administration of the allergen and BPPcysMPEG enhanced expression of co-stimulatory molecules on DC and increased secretion of the proinflammatory cytokine TNF-α. Acting as an adjuvant, BPPcysMPEG elevated allergen-specific immune responses in co-culture with autologous lymphocytes. Although administration of BPPcysMPEG alone enhanced expression of co-stimulatory molecules on DC, proliferation of autologous lymphocytes was not induced. Based on this finding, the potential of BPPcysMPEG to reduce allergic airway inflammation by preventive modulation of the innate immune system via TLR2/6 agonization was investigated in mice. Local administration of BPPcysMPEG altered cellular influx and cell composition in BAL fluid. Furthermore, the Th2-associated cytokines IL-4 and IL-5 were diminished. Allergen-specific restimulation of cells from mediastinal lymph nodes and splenocytes suggested an alteration of immune responses. The treatment with BPPcysMPEG induced a Th1-dominated cytokine milieu in mediastinal lymph nodes, while allergen-specific immune responses in splenocytes were diminished. The co-administration of allergen and BPPcysMPEG reduced cytokine secretion upon restimulation in mediastinal lymph nodes and splenocytes. From these data we conclude that BPPcysMPEG was able to influence the immune system with regard to subsequent allergen contact by TLR2/6 agonization.
2011-07-07T14:19:36Z
2011-07-07T14:19:36Z
2011-07-07T14:19:36Z
Article
Local treatment with BPPcysMPEG reduces allergic airway inflammation in sensitized mice., 216 (1-2):110-7 Immunobiology
1878-3279
20619481
10.1016/j.imbio.2010.05.003
http://hdl.handle.net/10033/135574
Immunobiology
en
oai:repository.helmholtz-hzi.de:10033/1405942019-08-30T11:37:23Zcom_10033_620652col_10033_620666
2011-08-24T12:11:03Z
urn:hdl:10033/140594
Peritoneal Cavity Is Dominated by IFNγ-Secreting CXCR3 Th1 Cells.
Zygmunt, Beata M
Groebe, Lothar
Guzman, Carlos A
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
The chemokine receptor CXCR3, which was shown to take part in many inflammatory processes, is considered as a Th1 specific marker. Here, we show in a mouse model that CXCR3 expressing CD4(+) cells preferentially migrate to the peritoneal cavity under steady-state conditions. The peritoneal cavity milieu leads to an up-regulated expression of CXCR3. However, blocking of known ligands of this chemokine receptor did not alter the preferential migration. The peritoneal cavity environment also results in an increased percentage of memory cells producing cytokines. Up-regulation of IFNγ production occurs mostly in CXCR3(+) cells considered as Th1, whereas the up-regulation of IL-4 affects mostly in CXCR3(-) cells which are considered as Th2. We conclude that the peritoneal cavity does not change the Th-lineage of the cells, but that domination of this anatomic niche by Th1 cells rather results from preferential migration to this compartment.
2011-08-24T12:11:03Z
2011-08-24T12:11:03Z
2011
Article
Peritoneal Cavity Is Dominated by IFNγ-Secreting CXCR3 Th1 Cells. 2011, 6 (7):e18032 PLoS ONE
1932-6203
21789162
10.1371/journal.pone.0018032
http://hdl.handle.net/10033/140594
PloS one
en
oai:repository.helmholtz-hzi.de:10033/1431792019-08-30T11:37:44Zcom_10033_620652col_10033_620666
2011-09-26T14:05:08Z
urn:hdl:10033/143179
Peritoneal cavity is dominated by IFNγ-secreting CXCR3+ Th1 cells.
Zygmunt, Beata M
Groebe, Lothar
Guzman, Carlos A
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
The chemokine receptor CXCR3, which was shown to take part in many inflammatory processes, is considered as a Th1 specific marker. Here, we show in a mouse model that CXCR3 expressing CD4(+) cells preferentially migrate to the peritoneal cavity under steady-state conditions. The peritoneal cavity milieu leads to an up-regulated expression of CXCR3. However, blocking of known ligands of this chemokine receptor did not alter the preferential migration. The peritoneal cavity environment also results in an increased percentage of memory cells producing cytokines. Up-regulation of IFNγ production occurs mostly in CXCR3(+) cells considered as Th1, whereas the up-regulation of IL-4 affects mostly in CXCR3(-) cells which are considered as Th2. We conclude that the peritoneal cavity does not change the Th-lineage of the cells, but that domination of this anatomic niche by Th1 cells rather results from preferential migration to this compartment.
2011-09-26T14:05:08Z
2011-09-26T14:05:08Z
2011
Article
Peritoneal cavity is dominated by IFNγ-secreting CXCR3+ Th1 cells. 2011, 6 (7):e18032 PLoS ONE
1932-6203
21789162
10.1371/journal.pone.0018032
http://hdl.handle.net/10033/143179
PloS one
en
oai:repository.helmholtz-hzi.de:10033/2104912019-08-30T11:30:32Zcom_10033_620652col_10033_620666
2012-02-10T15:36:17Z
urn:hdl:10033/210491
Pidotimod promotes functional maturation of dendritic cells and displays adjuvant properties at the nasal mucosa level.
Giagulli, Cinzia
Noerder, Miriam
Avolio, Manuela
Becker, Pablo D
Fiorentini, Simona
Guzman, Carlos A
Caruso, Arnaldo
Department of Experimental and Applied Medicine, Section of Microbiology, University of Brescia, Medical School, Brescia, Italy.
Mucosal dendritic cells (DCs) are very important in the process of antigen presentation to T cells, playing a key role in the induction of primary and secondary immune responses. Pidotimod is a synthetic substance capable of modulating immune cell functions, but the effect of pidotimod on human DCs has not been investigated yet. Here we demonstrate the ability of pidotimod to induce DC maturation and up-regulate the expression of HLA-DR and co-stimulatory molecules CD83 and CD86, which are fundamental for communication with adaptative immunity cells. Pidotimod also stimulated DCs to release high amounts of pro-inflammatory molecules such as MCP-1 and TNF-alpha cytokines and to drive T cell proliferation and differentiation towards a Th1 phenotype. Moreover, we demonstrate that pidotimod in vivo promotes strong and specific humoral and cellular immune response when co-administered intranasally with a model antigen. Taken together our data suggest the possibility to use pidotimod as adjuvant molecule to facilitate the activation of the innate immune system as well as to promote an effective mucosal and systemic immune response.
2012-02-10T15:36:17Z
2012-02-10T15:36:17Z
2009-11
Article
Pidotimod promotes functional maturation of dendritic cells and displays adjuvant properties at the nasal mucosa level. 2009, 9 (12):1366-73 Int. Immunopharmacol.
1878-1705
19712757
10.1016/j.intimp.2009.08.010
http://hdl.handle.net/10033/210491
International immunopharmacology
en
oai:repository.helmholtz-hzi.de:10033/2168642019-08-30T11:35:13Zcom_10033_620652col_10033_620666
2012-03-28T13:19:57Z
urn:hdl:10033/216864
Gene expression profiles of T cells from hepatitis E virus infected patients in acute and resolving phase.
TrehanPati, Nirupma
Sukriti, Sukriti
Geffers, Robert
Hissar, Syed
Riese, Peggy
Toepfer, Tanja
Guzman, Carlos A
Sarin, Shiv Kumar
Institute of Liver and Biliary Sciences, D-1, Vasant Kunj, 110 070, New Delhi, India. trehanpati@gmail.com
Approximately 50% of acute viral hepatitis in young adults and in pregnant women is due to hepatitis E virus (HEV) infection in developing countries. T cell-mediated immune injury probably plays a key role in the pathogenesis of acute hepatitis illness. However, there is a paucity of data on the global gene expression programs activated on T cells, which are subsequently responsible for T cell recruitment to the liver and triggering of immune injury.
2012-03-28T13:19:57Z
2012-03-28T13:19:57Z
2011-06
Article
Gene expression profiles of T cells from hepatitis E virus infected patients in acute and resolving phase. 2011, 31 (3):498-508 J. Clin. Immunol.
1573-2592
21287396
10.1007/s10875-010-9506-2
http://hdl.handle.net/10033/216864
Journal of clinical immunology
en
Archived with thanks to Journal of clinical immunology
oai:repository.helmholtz-hzi.de:10033/2255832019-08-30T11:36:05Zcom_10033_620652col_10033_620666
2012-05-23T13:13:50Z
urn:hdl:10033/225583
Cyanoethylation of the glucans dextran and pullulan: Substitution pattern and formation of nanostructures and entrapment of magnetic nanoparticles.
Fiege, Kathrin
Lünsdorf, Heinrich
Atarijabarzadeh, Sevil
Mischnick, Petra
Institute for Food Chemistry, Technische Universität Braunschweig, Schleinitzstraße 20, D-38106 Braunschweig, Germany.
Cyanoethylglucans with a degree of substitution in the range of 0.74 to 2.40 for dextran and 0.84 to 2.42 for pullulan were obtained by Michael addition of acrylonitrile to the glucans under various conditions. Products were thoroughly characterized, comprising elementary analysis, NMR and ATR-IR spectroscopy, and analysis of the substituent distribution in the glucosyl units by GC-FID and GC-MS of the constituting monosaccharide derivatives. Nanostructuring of the highly substituted cyanoethylpolysaccharides was performed by dialysis against a non-solvent. In the presence of ferromagnetic iron-oxide nanoparticles, multicore cyanoethylglucan-coated ferromagnetic nanoparticles were formed by selective entrapment. The specific interaction between cyano groups and iron could be proven. The size distribution and morphology of the nanoparticles were analyzed by dynamic light scattering (DLS), scanning electron microscopy (SEM) and energy-filtered transmission electron microscopy (EF-TEM) with parallel electron energy loss spectroscopy (PEELS).
2012-05-23T13:13:50Z
2012-05-23T13:13:50Z
2012
Article
Cyanoethylation of the glucans dextran and pullulan: Substitution pattern and formation of nanostructures and entrapment of magnetic nanoparticles. 2012, 8:551-66 Beilstein J Org Chem
1860-5397
22563354
10.3762/bjoc.8.63
http://hdl.handle.net/10033/225583
Beilstein journal of organic chemistry
en
Archived with thanks to Beilstein journal of organic chemistry
oai:repository.helmholtz-hzi.de:10033/2296352019-08-30T11:36:33Zcom_10033_620652col_10033_620666
2012-06-19T11:40:47Z
urn:hdl:10033/229635
NKT cell stimulation with α-galactosylceramide results in a block of Th17 differentiation after intranasal immunization in mice.
Zygmunt, Beata M
Weissmann, Sebastian F
Guzman, Carlos A
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany. beata.zygmunt@bbsrc.ac.uk
In a previous study we demonstrated that intranasal (i.n.) vaccination promotes a Th17 biased immune response. Here, we show that co-administration of a pegylated derivative of α-galactosylceramide (αGCPEG) with an antigen, even in the presence of Th17-polarizing compounds, results in a strong blocking of Th17 differentiation. Additional studies demonstrated that this phenomenon is specifically dependent on soluble factors, like IL-4 and IFNγ, which are produced by NKT cells. Even NK1.1 negative NKT cells, which by themselves produce IL-17A, are able to block Th17 differentiation. It follows that the use of αGCPEG as adjuvant would enable to tailor Th17 responses, according to the specific clinical needs. This knowledge expands our understanding of the role played by NKT cells in overall control of the cytokine microenvironment, as well as in the overall shaping of adaptive immune responses.
2012-06-19T11:40:47Z
2012-06-19T11:40:47Z
2012
Article
NKT cell stimulation with α-galactosylceramide results in a block of Th17 differentiation after intranasal immunization in mice. 2012, 7 (1):e30382 PLoS ONE
1932-6203
22291945
10.1371/journal.pone.0030382
http://hdl.handle.net/10033/229635
PloS one
en
Archived with thanks to PloS one
oai:repository.helmholtz-hzi.de:10033/2318512019-08-30T11:25:43Zcom_10033_620652col_10033_620666
2012-07-03T08:22:53Z
urn:hdl:10033/231851
Long-term effects of ocean warming on the prokaryotic community: evidence from the vibrios.
Vezzulli, Luigi
Brettar, Ingrid
Pezzati, Elisabetta
Reid, Philip C
Colwell, Rita R
Höfle, Manfred G
Pruzzo, Carla
Department for the Study of Territory and its Resources, University of Genoa, Genoa, Italy. luigi.vezzulli@unige.it
The long-term effects of ocean warming on prokaryotic communities are unknown because of lack of historical data. We overcame this gap by applying a retrospective molecular analysis to the bacterial community on formalin-fixed samples from the historical Continuous Plankton Recorder archive, which is one of the longest and most geographically extensive collections of marine biological samples in the world. We showed that during the last half century, ubiquitous marine bacteria of the Vibrio genus, including Vibrio cholerae, increased in dominance within the plankton-associated bacterial community of the North Sea, where an unprecedented increase in bathing infections related to these bacteria was recently reported. Among environmental variables, increased sea surface temperature explained 45% of the variance in Vibrio data, supporting the view that ocean warming is favouring the spread of vibrios and may be the cause of the globally increasing trend in their associated diseases.
2012-07-03T08:22:53Z
2012-07-03T08:22:53Z
2012-01
Article
Long-term effects of ocean warming on the prokaryotic community: evidence from the vibrios. 2012, 6 (1):21-30 ISME J
1751-7370
21753799
10.1038/ismej.2011.89
http://hdl.handle.net/10033/231851
The ISME journal
en
Archived with thanks to The ISME journal
oai:repository.helmholtz-hzi.de:10033/2358312019-08-30T11:37:00Zcom_10033_620652col_10033_620666
2012-07-26T08:18:45Z
urn:hdl:10033/235831
Evaluation of the sublingual route for administration of influenza H5N1 virosomes in combination with the bacterial second messenger c-di-GMP.
Pedersen, Gabriel Kristian
Ebensen, Thomas
Gjeraker, Ingrid Hjetland
Svindland, Signe
Bredholt, Geir
Guzmán, Carlos Alberto
Cox, Rebecca Jane
The Gade Institute, University of Bergen, Norway. gabriel.pedersen@gades.uib.no
Avian influenza A H5N1 is a virus with pandemic potential. Mucosal vaccines are attractive as they have the potential to block viruses at the site of entry, thereby preventing both disease and further transmission. The intranasal route is safe for the administration of seasonal live-attenuated influenza vaccines, but may be less suitable for administration of pandemic vaccines. Research into novel mucosal routes is therefore needed. In this study, a murine model was used to compare sublingual administration with intranasal and intramuscular administration of influenza H5N1 virosomes (2 µg haemagglutinin; HA) in combination with the mucosal adjuvant (3',5')-cyclic dimeric guanylic acid (c-di-GMP). We found that sublingual immunisation effectively induced local and systemic H5N1-specific humoral and cellular immune responses but that the magnitude of response was lower than after intranasal administration. However, both the mucosal routes were superior to intramuscular immunisation for induction of local humoral and systemic cellular immune responses including high frequencies of splenic H5N1-specific multifunctional (IL-2+TNF-α+) CD4+ T cells. The c-di-GMP adjuvanted vaccine elicited systemic haemagglutination inhibition (HI) antibody responses (geometric mean titres ≥ 40) both when administered sublingually, intranasally and inramuscularly. In addition, salivary HI antibodies were elicited by mucosal, but not intramuscular vaccination. We conclude that the sublingual route is an attractive alternative for administration of pandemic influenza vaccines.
2012-07-26T08:18:45Z
2012-07-26T08:18:45Z
2011
Article
Evaluation of the sublingual route for administration of influenza H5N1 virosomes in combination with the bacterial second messenger c-di-GMP. 2011, 6 (11):e26973 PLoS ONE
1932-6203
22069479
10.1371/journal.pone.0026973
http://hdl.handle.net/10033/235831
PloS one
en
Archived with thanks to PloS one
oai:repository.helmholtz-hzi.de:10033/2445912019-08-30T11:28:24Zcom_10033_620652col_10033_620666
2012-09-18T08:22:06Z
urn:hdl:10033/244591
Assessing the viability of bacterial species in drinking water by combined cellular and molecular analyses.
Kahlisch, Leila
Henne, Karsten
Gröbe, Lothar
Brettar, Ingrid
Höfle, Manfred G
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany.
The question which bacterial species are present in water and if they are viable is essential for drinking water safety but also of general relevance in aquatic ecology. To approach this question we combined propidium iodide/SYTO9 staining ("live/dead staining" indicating membrane integrity), fluorescence-activated cell sorting (FACS) and community fingerprinting for the analysis of a set of tap water samples. Live/dead staining revealed that about half of the bacteria in the tap water had intact membranes. Molecular analysis using 16S rRNA and 16S rRNA gene-based single-strand conformation polymorphism (SSCP) fingerprints and sequencing of drinking water bacteria before and after FACS sorting revealed: (1) the DNA- and RNA-based overall community structure differed substantially, (2) the community retrieved from RNA and DNA reflected different bacterial species, classified as 53 phylotypes (with only two common phylotypes), (3) the percentage of phylotypes with intact membranes or damaged cells were comparable for RNA- and DNA-based analyses, and (4) the retrieved species were primarily of aquatic origin. The pronounced difference between phylotypes obtained from DNA extracts (dominated by Betaproteobacteria, Bacteroidetes, and Actinobacteria) and from RNA extracts (dominated by Alpha-, Beta-, Gammaproteobacteria, Bacteroidetes, and Cyanobacteria) demonstrate the relevance of concomitant RNA and DNA analyses for drinking water studies. Unexpected was that a comparable fraction (about 21%) of phylotypes with membrane-injured cells was observed for DNA- and RNA-based analyses, contradicting the current understanding that RNA-based analyses represent the actively growing fraction of the bacterial community. Overall, we think that this combined approach provides an interesting tool for a concomitant phylogenetic and viability analysis of bacterial species of drinking water.
2012-09-18T08:22:06Z
2012-09-18T08:22:06Z
2012-02
Article
Assessing the viability of bacterial species in drinking water by combined cellular and molecular analyses. 2012, 63 (2):383-97 Microb. Ecol.
1432-184X
21845446
10.1007/s00248-011-9918-4
http://hdl.handle.net/10033/244591
Microbial ecology
en
Archived with thanks to Microbial ecology
oai:repository.helmholtz-hzi.de:10033/2445922019-08-30T11:28:24Zcom_10033_620652col_10033_620666
2012-09-18T08:34:00Z
urn:hdl:10033/244592
Analysis of bacterial core communities in the central Baltic by comparative RNA-DNA-based fingerprinting provides links to structure-function relationships.
Brettar, Ingrid
Christen, Richard
Höfle, Manfred G
Department of Vaccinology and Applied Microbiology, Helmholtz Centre of Infection Research (HZI), Braunschweig, Germany.
Understanding structure-function links of microbial communities is a central theme of microbial ecology since its beginning. To this end, we studied the spatial variability of the bacterioplankton community structure and composition across the central Baltic Sea at four stations, which were up to 450 km apart and at a depth profile representative for the central part (Gotland Deep, 235 m). Bacterial community structure was followed by 16S ribosomal RNA (rRNA)- and 16S rRNA gene-based fingerprints using single-strand conformation polymorphism (SSCP) electrophoresis. Species composition was determined by sequence analysis of SSCP bands. High similarities of the bacterioplankton communities across several hundred kilometers were observed in the surface water using RNA- and DNA-based fingerprints. In these surface communities, the RNA- and DNA-based fingerprints resulted in very different pattern, presumably indicating large difference between the active members of the community as represented by RNA-based fingerprints and the present members represented by the DNA-based fingerprints. This large discrepancy changed gradually over depth, resulting in highly similar RNA- and DNA-based fingerprints in the anoxic part of the water column below 130 m depth. A conceivable mechanism explaining this high similarity could be the reduced oxidative stress in the anoxic zone. The stable communities on the surface and in the anoxic zone indicate the strong influence of the hydrography on the bacterioplankton community structure. Comparative analysis of RNA- and DNA-based community structure provided criteria for the identification of the core community, its key members and their links to biogeochemical functions.
2012-09-18T08:34:00Z
2012-09-18T08:34:00Z
2012-01
Article
Analysis of bacterial core communities in the central Baltic by comparative RNA-DNA-based fingerprinting provides links to structure-function relationships. 2012, 6 (1):195-212 ISME J
1751-7370
21697960
10.1038/ismej.2011.80
http://hdl.handle.net/10033/244592
The ISME journal
en
Archived with thanks to The ISME journal
oai:repository.helmholtz-hzi.de:10033/2452312019-08-30T11:28:51Zcom_10033_620652col_10033_620666
2012-09-20T11:17:16Z
urn:hdl:10033/245231
Microevolution of pandemic Vibrio parahaemolyticus assessed by the number of repeat units in short sequence tandem repeat regions.
García, Katherine
Gavilán, Ronnie G
Höfle, Manfred G
Martínez-Urtaza, Jaime
Espejo, Romilio T
Instituto de Nutrición y Tecnología de los Alimentos, Universidad de Chile, Santiago, Chile.
The emergence of the pandemic strain Vibrio parahaemolyticus O3:K6 in 1996 caused a large increase of diarrhea outbreaks related to seafood consumption in Southeast Asia, and later worldwide. Isolates of this strain constitutes a clonal complex, and their effectual differentiation is possible by comparison of their variable number tandem repeats (VNTRs). The differentiation of the isolates by the differences in VNTRs will allow inferring the population dynamics and microevolution of this strain but this requires knowing the rate and mechanism of VNTRs' variation. Our study of mutants obtained after serial cultivation of clones showed that mutation rates of the six VNTRs examined are on the order of 10(-4) mutant per generation and that difference increases by stepwise addition of single mutations. The single stepwise mutation (SSM) was deduced because mutants with 1, 2, 3, or more repeat unit deletions or insertions follow a geometric distribution. Plausible phylogenetic trees are obtained when, according to SSM, the genetic distance between clusters with different number of repeats is assessed by the absolute differences in repeats. Using this approach, mutants originated from different isolates of pandemic V. parahaemolyticus after serial cultivation are clustered with their parental isolates. Additionally, isolates of pandemic V. parahaemolyticus from Southeast Asia, Tokyo, and northern and southern Chile are clustered according their geographical origin. The deepest split in these four populations is observed between the Tokyo and southern Chile populations. We conclude that proper phylogenetic relations and successful tracing of pandemic V. parahaemolyticus requires measuring the differences between isolates by the absolute number of repeats in the VNTRs considered.
2012-09-20T11:17:16Z
2012-09-20T11:17:16Z
2012
Article
Microevolution of pandemic Vibrio parahaemolyticus assessed by the number of repeat units in short sequence tandem repeat regions. 2012, 7 (1):e30823 PLoS ONE
1932-6203
22292049
10.1371/journal.pone.0030823
http://hdl.handle.net/10033/245231
PloS one
en
Archived with thanks to PloS one
oai:repository.helmholtz-hzi.de:10033/2541722019-08-30T11:28:51Zcom_10033_620652col_10033_620666
2012-11-30T20:32:40Z
urn:hdl:10033/254172
NKT cell stimulation with α-galactosylceramide results in a block of Th17 differentiation after intranasal immunization in mice.
Zygmunt, Beata M
Weissmann, Sebastian F
Guzman, Carlos A
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany. beata.zygmunt@bbsrc.ac.uk
In a previous study we demonstrated that intranasal (i.n.) vaccination promotes a Th17 biased immune response. Here, we show that co-administration of a pegylated derivative of α-galactosylceramide (αGCPEG) with an antigen, even in the presence of Th17-polarizing compounds, results in a strong blocking of Th17 differentiation. Additional studies demonstrated that this phenomenon is specifically dependent on soluble factors, like IL-4 and IFNγ, which are produced by NKT cells. Even NK1.1 negative NKT cells, which by themselves produce IL-17A, are able to block Th17 differentiation. It follows that the use of αGCPEG as adjuvant would enable to tailor Th17 responses, according to the specific clinical needs. This knowledge expands our understanding of the role played by NKT cells in overall control of the cytokine microenvironment, as well as in the overall shaping of adaptive immune responses.
2012-11-30T20:32:40Z
2012-11-30T20:32:40Z
2012
Article
NKT cell stimulation with α-galactosylceramide results in a block of Th17 differentiation after intranasal immunization in mice. 2012, 7 (1):e30382 PLoS ONE
1932-6203
22291945
10.1371/journal.pone.0030382
http://hdl.handle.net/10033/254172
PloS one
en
Archived with thanks to PloS one
oai:repository.helmholtz-hzi.de:10033/2552432019-08-30T11:28:51Zcom_10033_620652col_10033_620666
2012-12-11T15:38:47Z
urn:hdl:10033/255243
Vaccines: from empirical development to rational design.
Rueckert, Christine
Guzmán, Carlos A
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
Infectious diseases are responsible for an overwhelming number of deaths worldwide and their clinical management is often hampered by the emergence of multi-drug-resistant strains. Therefore, prevention through vaccination currently represents the best course of action to combat them. However, immune escape and evasion by pathogens often render vaccine development difficult. Furthermore, most currently available vaccines were empirically designed. In this review, we discuss why rational design of vaccines is not only desirable but also necessary. We introduce recent developments towards specifically tailored antigens, adjuvants, and delivery systems, and discuss the methodological gaps and lack of knowledge still hampering true rational vaccine design. Finally, we address the potential and limitations of different strategies and technologies for advancing vaccine development.
2012-12-11T15:38:47Z
2012-12-11T15:38:47Z
2012-11
Article
Vaccines: from empirical development to rational design. 2012, 8 (11):e1003001 PLoS Pathog.
1553-7374
23144616
10.1371/journal.ppat.1003001
http://hdl.handle.net/10033/255243
PLoS pathogens
en
Archived with thanks to PLoS pathogens
oai:repository.helmholtz-hzi.de:10033/2837792019-08-30T11:30:32Zcom_10033_620652col_10033_620666
2013-04-24T14:32:39Z
urn:hdl:10033/283779
Development and characterization of attenuated metabolic mutants of Bordetella bronchiseptica for applications in vaccinology.
Yevsa, Tetyana
Ebensen, Thomas
Fuchs, Barbara
Zygmunt, Beata
Libanova, Rimma
Gross, Roy
Schulze, Kai
Guzmán, Carlos A
Department of Vaccinology and Applied Microbiology, Helmholtz Centre for Infection Research, D-38124 Braunschweig, Germany.
Bordetella bronchiseptica is an important pathogen causing a number of veterinary respiratory syndromes in agriculturally important and food-producing confinement-reared animals, resulting in great economic losses annually amounting to billions of euros worldwide. Currently available live vaccines are incompletely satisfactory in terms of efficacy and safety. An efficient vaccine for livestock animals would allow reducing the application of antibiotics, thereby preventing the massive release of pharmaceuticals into the environment. Here, we describe two new potential vaccine strains based on the BB7865 strain. Two independent attenuating mutations were incorporated by homologous recombination in order to make negligible the risk of recombination and subsequent reversion to the virulent phenotype. The mutations are critical for bacterial metabolism, resistance to oxidative stress, intracellular survival and in vivo persistence. The resulting double mutants BB7865 risA aroA and BB7865 risA dapE were characterized as promising vaccine candidates, which are able to confer protection against colonization of the lower respiratory tract after sublethal challenge with the wild-type strain.
2013-04-24T14:32:39Z
2013-04-24T14:32:39Z
2013-01
Article
Development and characterization of attenuated metabolic mutants of Bordetella bronchiseptica for applications in vaccinology. 2013, 15 (1):64-76 Environ. Microbiol.
1462-2920
22676396
10.1111/j.1462-2920.2012.02779.x
http://hdl.handle.net/10033/283779
Environmental microbiology
en
Archived with thanks to Environmental microbiology
oai:repository.helmholtz-hzi.de:10033/3112972019-08-30T11:37:24Zcom_10033_620652col_10033_620666
2014-01-14T15:11:57Z
urn:hdl:10033/311297
A study of Chitosan and c-di-GMP as mucosal adjuvants for intranasal influenza H5N1 vaccine.
Svindland, Signe C
Pedersen, Gabriel K
Pathirana, Rishi D
Bredholt, Geir
Nøstbakken, Jane K
Jul-Larsen, Åsne
Guzmán, Carlos A
Montomoli, Emanuele
Lapini, Giulia
Piccirella, Simona
Jabbal-Gill, Inderjit
Hinchcliffe, Michael
Cox, Rebecca J
department of vaccinology and applied microbiology, Helmholtz Centre for infection research, D38124 Braunschweig, Germany
Highly pathogenic avian influenza A/H5N1 virus remains a potential pandemic threat, and it is essential to continue vaccine development against this subtype. A local mucosal immune response in the upper respiratory tract may stop influenza transmission. It is therefore important to develop effective intranasal pandemic influenza vaccines that induce mucosal immunity at the site of viral entry.
2014-01-14T15:11:57Z
2014-01-14T15:11:57Z
2013-11
Article
A study of Chitosan and c-di-GMP as mucosal adjuvants for intranasal influenza H5N1 vaccine. 2013, 7 (6):1181-93 Influenza Other Respir Viruses
1750-2659
23170900
10.1111/irv.12056
http://hdl.handle.net/10033/311297
Influenza and other respiratory viruses
en
Archived with thanks to Influenza and other respiratory viruses
oai:repository.helmholtz-hzi.de:10033/3116222019-08-30T11:27:46Zcom_10033_620652col_10033_620666
2014-01-20T14:17:27Z
urn:hdl:10033/311622
Dynamic changes in viral population structure and compartmentalization during chronic hepatitis C virus infection in children.
Gismondi, María Inés
Díaz Carrasco, Juan María
Valva, Pamela
Becker, Pablo Daniel
Guzmán, Carlos Alberto
Campos, Rodolfo Héctor
Preciado, María Victoria
Helmholtz Centre for infection research, D-38124 Braunschweig, Germany
Classic phylogenetic and modern population-based clustering methods were used to analyze hepatitis C virus (HCV) evolution in plasma and to assess viral compartmentalization within peripheral blood mononuclear cells (PBMCs) in 6 children during 3.2-9.6yr of follow-up. Population structure analysis of cloned amplicons encompassing hypervariable region 1 led to the distinction of two evolutionary patterns, one highly divergent and another one genetically homogeneous. Viral adaptability was reflected by co-evolution of viral communities switching rapidly from one to another in the context of divergence and stability associated with highly homogeneous communities which were replaced by new ones after long periods. Additionally, viral compartmentalization of HCV in PBMCs was statistically demonstrated, suggesting their role as a pool of genetic variability. Our results support the idea of a community-based structure of HCV viral populations during chronic infection and highlight a role of the PBMC compartment in the persistence of such structure.
2014-01-20T14:17:27Z
2014-01-20T14:17:27Z
2013-12
Article
Dynamic changes in viral population structure and compartmentalization during chronic hepatitis C virus infection in children. 2013, 447 (1-2):187-96 Virology
1096-0341
24210114
10.1016/j.virol.2013.09.002
http://hdl.handle.net/10033/311622
Virology
en
Archived with thanks to Virology
oai:repository.helmholtz-hzi.de:10033/3128722019-08-30T11:33:29Zcom_10033_620652col_10033_620666
2014-02-14T14:54:49Z
urn:hdl:10033/312872
Seasonal dynamics of bacterial community structure and composition in cold and hot drinking water derived from surface water reservoirs.
Henne, Karsten
Kahlisch, Leila
Höfle, Manfred G
Brettar, Ingrid
Dept of Vacciology, Helmholtz Centre for infection research, D-38124 Braunschweig, Germany
In temperate regions, seasonal variability of environmental factors affects the bacterial community in source water and finished drinking water. Therefore, the bacterial core community and its seasonal variability in cold and the respective hot drinking water was investigated. The bacterial core community was studied by 16S rRNA-based SSCP fingerprint analyses and band sequencing of DNA and RNA extracts of cold and hot water (60 °C). The bacterial communities of cold and hot drinking water showed a highly different structure and phylogenetic composition both for RNA and DNA extracts. For cold drinking water substantial seasonal dynamics of the bacterial community was observed related to environmental factors such as temperature and precipitation affecting source and drinking water. Phylogenetic analyses of the cold water community indicated that the majority of phylotypes were very closely affiliated with those detected in former studies of the same drinking water supply system (DWSS) in the preceding 6 years, indicating a high stability over time. The hot water community was very stable over time and seasons and highly distinct from the cold water with respect to structure and composition. The hot water community displayed a lower diversity and its phylotypes were mostly affiliated with bacteria of high temperature habitats with high growth rates indicated by their high RNA content. The conversion of the cold to the hot water bacterial community is considered as occurring within a few hours by the following two processes, i) by decay of most of the cold water bacteria due to heating, and ii) rapid growth of the high temperature adapted bacteria present in the hot water (co-heated with the cold water in the same device) using the nutrients released from the decaying cold water bacteria. The high temperature adapted bacteria originated partially from low abundant but beforehand detected members of the cold water; additionally, the rare members ("seed bank ") of the cold water are considered as a source.
2014-02-14T14:54:49Z
2014-02-14T14:54:49Z
2014-02-14
Article
Seasonal dynamics of bacterial community structure and composition in cold and hot drinking water derived from surface water reservoirs. 2013, 47 (15):5614-30 Water Res.
1879-2448
23890873
10.1016/j.watres.2013.06.034
http://hdl.handle.net/10033/312872
Water research
en
Archived with thanks to Water research
oai:repository.helmholtz-hzi.de:10033/3172602019-08-30T11:32:17Zcom_10033_620652col_10033_620666
2014-05-21T13:59:10Z
urn:hdl:10033/317260
The Mucosal Adjuvant Cyclic di-AMP Exerts Immune Stimulatory Effects on Dendritic Cells and Macrophages.
Skrnjug, Ivana
Rueckert, Christine
Libanova, Rimma
Lienenklaus, Stefan
Weiss, Siegfried
Guzmán, Carlos A
The cyclic di-nucleotide bis-(3',5')-cyclic dimeric adenosine monophosphate (c-di-AMP) is a candidate mucosal adjuvant with proven efficacy in preclinical models. It was shown to promote specific humoral and cellular immune responses following mucosal administration. To date, there is only fragmentary knowledge on the cellular and molecular mode of action of c-di-AMP. Here, we report on the identification of dendritic cells and macrophages as target cells of c-di-AMP. We show that c-di-AMP induces the cell surface up-regulation of T cell co-stimulatory molecules as well as the production of interferon-β. Those responses were characterized by in vitro experiments with murine and human immune cells and in vivo studies in mice. Analyses of dendritic cell subsets revealed conventional dendritic cells as principal responders to stimulation by c-di-AMP. We discuss the impact of the reported antigen presenting cell activation on the previously observed adjuvant effects of c-di-AMP in mouse immunization studies.
2014-05-21T13:59:10Z
2014-05-21T13:59:10Z
2014
Article
The Mucosal Adjuvant Cyclic di-AMP Exerts Immune Stimulatory Effects on Dendritic Cells and Macrophages. 2014, 9 (4):e95728 PLoS ONE
1932-6203
24755640
10.1371/journal.pone.0095728
http://hdl.handle.net/10033/317260
PloS one
en
Archived with thanks to PloS one
oai:repository.helmholtz-hzi.de:10033/3208112019-08-30T11:35:13Zcom_10033_620652col_10033_620666
2014-06-11T14:30:02Z
urn:hdl:10033/320811
Analysis of structure and composition of bacterial core communities in mature drinking water biofilms and bulk water of a citywide network in Germany.
Henne, Karsten
Kahlisch, Leila
Brettar, Ingrid
Höfle, Manfred G
The bacterial core communities of bulk water and corresponding biofilms of a more than 20-year-old drinking water network were compared using 16S rRNA single-strand confirmation polymorphism (SSCP) fingerprints based on extracted DNA and RNA. The structure and composition of the bacterial core community in the bulk water was highly similar (>70%) across the city of Braunschweig, Germany, whereas all biofilm samples contained a unique community with no overlapping phylotypes from bulk water. Biofilm samples consisted mainly of Alphaproteobacteria (26% of all phylotypes), Gammaproteobacteria (11%), candidate division TM6 (11%), Chlamydiales (9%), and Betaproteobacteria (9%). The bulk water community consisted primarily of Bacteroidetes (25%), Betaproteobacteria (20%), Actinobacteria (16%), and Alphaproteobacteria (11%). All biofilm communities showed higher relative abundances of single phylotypes and a reduced richness compared to bulk water. Only biofilm communities sampled at nearby sampling points showed similar communities irrespective of support materials. In all of our bulk water studies, the community composition determined from 16S rRNA was completely different from the 16S rRNA gene-based community composition, whereas in biofilms both molecular fractions resulted in community compositions that were similar to each other. We hypothesize that a higher fraction of active bacterial phylotypes and a better protection from oxidative stress in drinking water biofilms are responsible for this higher similarity.
2014-06-11T14:30:02Z
2014-06-11T14:30:02Z
2012-05
Article
Analysis of structure and composition of bacterial core communities in mature drinking water biofilms and bulk water of a citywide network in Germany. 2012, 78 (10):3530-8 Appl. Environ. Microbiol.
1098-5336
22389373
10.1128/AEM.06373-11
http://hdl.handle.net/10033/320811
Applied and environmental microbiology
en
Archived with thanks to Applied and environmental microbiology
oai:repository.helmholtz-hzi.de:10033/3221872019-08-30T11:25:11Zcom_10033_620652col_10033_620666
2014-06-23T14:23:39Z
urn:hdl:10033/322187
A new adjuvanted nanoparticle-based H1N1 influenza vaccine induced antigen-specific local mucosal and systemic immune responses after administration into the lung.
Neuhaus, Vanessa
Chichester, Jessica A
Ebensen, Thomas
Schwarz, Katharina
Hartman, Caitlin E
Shoji, Yoko
Guzmán, Carlos A
Yusibov, Vidadi
Sewald, Katherina
Braun, Armin
Annually influenza virus infections are responsible for hospitalization and mortality, especially in high risk groups. Constant antigenic changes in seasonal influenza viruses resulted from antigenic shifts and antigenic drifts, enable emerging of novel virus subtypes that may reduce current vaccine efficacy and impose the continuous revision of vaccine component. Currently available vaccines are usually limited by their production processes in terms of rapid adaptation to new circulating subtypes in high quantities meeting the global demand. Thus, new approaches to rapidly manufacture high yields of influenza vaccines are required. New technologies to reach maximal protection with minimal vaccine doses also need to be developed. In this study, we evaluated the systemic and local immunogenicity of a new double-adjuvanted influenza vaccine administered at the site of infection, the respiratory tract. This vaccine combines a plant-produced H1N1 influenza hemagglutinin antigen (HAC1), a silica nanoparticle-based (SiO₂) drug delivery system and the mucosal adjuvant candidate bis-(3',5')-cyclic dimeric guanosine monophosphate (c-di-GMP). Mice were vaccinated by intratracheal route with HAC1/SiO₂ or HAC1/c-di-GMP (single-adjuvanted vaccine) or HAC1/SiO₂/c-di-GMP (double-adjuvanted vaccine) and evaluated for target-specific immune responses, such as hemagglutination inhibition and hemagglutinin-specific IgG titers, as well as local antibody (IgG and IgA) titers in the bronchoalveolar lavage (BAL). Furthermore, the HAC1-specific T-cell re-stimulation potential was assessed using precision-cut lung slices (PCLS) of vaccinated mice. The double-adjuvanted vaccine induced high systemic antibody responses comparable to the systemic vaccination control. In addition, it induced local IgG and IgA responses in the BAL. Furthermore, HAC1 induced a local T-cell response demonstrated by elevated IL-2 and IFN-γ levels in PCLS of c-di-GMP-vaccinated mice upon re-stimulation. Overall, the present study showed the potential of the double-adjuvanted vaccine to induce systemic humoral immune responses in intratracheally vaccinated mice. Furthermore, it induced a strong mucosal immune response, with evidence of antigen-primed T-cells in the lung.
2014-06-23T14:23:39Z
2014-06-23T14:23:39Z
2014-05-30
Article
A new adjuvanted nanoparticle-based H1N1 influenza vaccine induced antigen-specific local mucosal and systemic immune responses after administration into the lung. 2014, 32 (26):3216-22 Vaccine
1873-2518
24731807
10.1016/j.vaccine.2014.04.011
http://hdl.handle.net/10033/322187
Vaccine
en
Archived with thanks to Vaccine
oai:repository.helmholtz-hzi.de:10033/3328412019-08-30T11:33:30Zcom_10033_620652col_10033_620666
2014-10-17T08:20:49Z
urn:hdl:10033/332841
Cyclic GMP-AMP Displays Mucosal Adjuvant Activity in Mice.
Skrnjug, Ivana
Guzmán, Carlos Alberto
Ruecker, Christine
Helmholtz Cente for infection research,Inhoffenstr. 7, D38124 Braunschweig, Germany.
The recently discovered mammalian enzyme cyclic GMP-AMP synthase produces cyclic GMP-AMP (cGAMP) after being activated by pathogen-derived cytosolic double stranded DNA. The product can stimulate STING-dependent interferon type I signaling. Here, we explore the efficacy of cGAMP as a mucosal adjuvant in mice. We show that cGAMP can enhance the adaptive immune response to the model antigen ovalbumin. It promotes antigen specific IgG and a balanced Th1/Th2 lymphocyte response in immunized mice. A characteristic of the cGAMP-induced immune response is the slightly reduced induction of interleukin-17 as a hallmark of Th17 activity - a distinct feature that is not observed with other cyclic di-nucleotide adjuvants. We further characterize the innate immune stimulation activity in vitro on murine bone marrow-derived dendritic cells and human dendritic cells. The observed results suggest the consideration of cGAMP as a candidate mucosal adjuvant for human vaccines.
2014-10-17T08:20:49Z
2014-10-17T08:20:49Z
2014
Article
Cyclic GMP-AMP Displays Mucosal Adjuvant Activity in Mice. 2014, 9 (10):e110150 PLoS ONE
1932-6203
25295996
10.1371/journal.pone.0110150
http://hdl.handle.net/10033/332841
PloS one
en
Archived with thanks to PloS one
oai:repository.helmholtz-hzi.de:10033/3440662019-08-30T11:37:44Zcom_10033_620652col_10033_620666
2015-02-03T14:49:25Z
urn:hdl:10033/344066
Use of S-[2,3-bispalmitoyiloxy-(2R)-propyl]-R-cysteinyl-amido-monomethoxy polyethylene glycol as an adjuvant improved protective immunity associated with a DNA vaccine encoding Cu,Zn superoxide dismutase of Brucella abortus in mice.
Retamal-Díaz, Angello
Riquelme-Neira, Roberto
Sáez, Darwin
Rivera, Alejandra
Fernández, Pablo
Cabrera, Alex
Guzmán, Carlos A
Oñate, Angel
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
This study was conducted to evaluate the immunogenicity and protective efficacy of a DNA vaccine encoding Brucella abortus Cu,Zn superoxide dismutase (SOD) using the Toll-like receptor 2/6 agonist S-[2,3-bispalmitoyiloxy-(2R)-propyl]-R-cysteinyl-amido-monomethoxy polyethylene glycol (BPPcysMPEG) as an adjuvant. Intranasal coadministration of BPPcysMPEG with a plasmid carrying the SOD-encoding gene (pcDNA-SOD) into BALB/c mice elicited antigen-specific humoral and cellular immune responses. Humoral responses were characterized by the stimulation of IgG2a and IgG1 and by the presence of SOD-specific secretory IgA in nasal and bronchoalveolar lavage fluids. Furthermore, T-cell proliferative responses and increased production of gamma interferon were also observed upon splenocyte restimulation with recombinant SOD. Cytotoxic responses were also stimulated, as demonstrated by the lysis of RB51-SOD-infected J774.A1 macrophages by cells recovered from immunized mice. The pcDNA-SOD/BPPcysMPEG formulation induced improved protection against challenge with the virulent strain B. abortus 2308 in BALB/c mice over that provided by pcDNA-SOD, suggesting the potential of this vaccination strategy against Brucella infection.
2015-02-03T14:49:25Z
2015-02-03T14:49:25Z
2014-11
Article
Use of S-[2,3-bispalmitoyiloxy-(2R)-propyl]-R-cysteinyl-amido-monomethoxy polyethylene glycol as an adjuvant improved protective immunity associated with a DNA vaccine encoding Cu,Zn superoxide dismutase of Brucella abortus in mice. 2014, 21 (11):1474-80 Clin. Vaccine Immunol.
1556-679X
25165025
10.1128/CVI.00554-14
http://hdl.handle.net/10033/344066
Clinical and vaccine immunology : CVI
en
oai:repository.helmholtz-hzi.de:10033/3443772019-08-30T11:28:23Zcom_10033_620652col_10033_620666
2015-02-11T15:49:31Z
urn:hdl:10033/344377
Efficient nanoparticle-mediated needle-free transcutaneous vaccination via hair follicles requires adjuvantation.
Mittal, Ankit
Schulze, Kai
Ebensen, Thomas
Weißmann, Sebastian
Hansen, Steffi
Lehr, Claus-Michael
Guzmán, Carlos A
Helmholtz Centre for Infection Research (HZI), Braunschweig, Germany.
Trans-follicular (TF) vaccination has recently been studied as a unique route for non-invasive transcutaneous vaccination. The present study aims to extensively characterize the immune responses triggered by TF vaccination using ovalbumin loaded chitosan-PLGA (poly lactic-co-glycolic acid) nanoparticles without skin pre-treatment to preserve skin integrity. The impact of formulation composition i.e. antigenic solution or antigen-loaded nanoparticles with or without adjuvant [bis-(3',5')-cyclic dimeric adenosine monophosphate] on immune response quality following TF immunization was analyzed and compared with immune responses obtained after tape stripping the skin. The results presented in this study confirm the ability of nanoparticle based vaccine formulations to deliver antigen across the intact skin via the follicular route, but at the same time demonstrate the necessity to include adjuvants to generate efficient antigen-specific humoral and cellular immune responses.
2015-02-11T15:49:31Z
2015-02-11T15:49:31Z
2015-01
Article
Efficient nanoparticle-mediated needle-free transcutaneous vaccination via hair follicles requires adjuvantation. 2015, 11 (1):147-54 Nanomedicine
1549-9642
25200611
10.1016/j.nano.2014.08.009
http://hdl.handle.net/10033/344377
Nanomedicine : nanotechnology, biology, and medicine
en
oai:repository.helmholtz-hzi.de:10033/5569942019-08-30T11:26:42Zcom_10033_620652col_10033_620666
2015-06-16T12:43:17Z
urn:hdl:10033/556994
Engineered dendritic cells from cord blood and adult blood accelerate effector T cell immune reconstitution against HCMV.
Daenthanasanmak, Anusara
Salguero, Gustavo
Sundarasetty, Bala Sai
Waskow, Claudia
Cosgun, Kadriye Nehir
Guzman, Carlos A
Riese, Peggy
Gerasch, Laura
Schneider, Andreas
Ingendoh, Alexandra
Messerle, Martin
Gabaev, Ildar
Woelk, Benno
Ruggiero, Eliana
Schmidt, Manfred
von Kalle, Christof
Figueiredo, Constanca
Eiz-Vesper, Britta
von Kaisenberg, Constantin
Ganser, Arnold
Stripecke, Renata
Helmholtz Centre for Infection Research, Inhoffenstraße 7, 38124 Braunschweig, Germany.
Human cytomegalovirus (HCMV) harmfully impacts survival after peripheral blood hematopoietic stem cell transplantation (PB-HSCT). Delayed immune reconstitution after cord blood (CB)-HSCT leads to even higher HCMV-related morbidity and mortality. Towards a feasible dendritic cell therapy to accelerate de novo immunity against HCMV, we validated a tricistronic integrase-defective lentiviral vector (coexpressing GM-CSF, IFN-α, and HCMV pp65 antigen) capable to directly induce self-differentiation of PB and CB monocytes into dendritic cells processing pp65 ("SmyleDCpp65"). In vitro, SmyleDCpp65 resisted HCMV infection, activated CD4(+) and CD8(+) T cells and expanded functional pp65-specific memory cytotoxic T lymphocytes (CTLs). CD34(+) cells obtained from PB and CB were transplanted into irradiated NOD.Rag1(-/-).IL2γc(-/-) mice. Donor-derived SmyleDCpp65 administration after PB-HSCT stimulated peripheral immune effects: lymph node remodeling, expansion of polyclonal effector memory CD8(+) T cells in blood, spleen and bone marrow, and pp65-reactive CTL and IgG responses. SmyleDCpp65 administration after CB-HSCT significantly stimulated thymopoiesis. Expanded frequencies of CD4(+)/CD8(+) T cell precursors containing increased levels of T-cell receptor excision circles in thymus correlated with peripheral expansion of effector memory CTL responses against pp65. The comparative in vivo modeling for PB and CB-HSCT provided dynamic and spatial information regarding human T and B cell reconstitution. In vivo potency supports future clinical development of SmyleDCpp65.
2015-06-16T12:43:17Z
2015-06-16T12:43:17Z
2015
Article
Engineered dendritic cells from cord blood and adult blood accelerate effector T cell immune reconstitution against HCMV. 2015, 1:14060 Mol Ther Methods Clin Dev
2329-0501
26052526
10.1038/mtm.2014.60
http://hdl.handle.net/10033/556994
Molecular therapy. Methods & clinical development
en
info:eu-repo/grantAgreement/EC/FP7/261387
openAccess
oai:repository.helmholtz-hzi.de:10033/5586982019-08-30T11:27:16Zcom_10033_620652col_10033_620666
2015-06-30T13:40:51Z
urn:hdl:10033/558698
TRANSVAC workshop on standardisation and harmonisation of analytical platforms for HIV, TB and malaria vaccines: 'how can big data help?'.
Dutruel, Céline
Thole, Jelle
Geels, Mark
Mollenkopf, Hans-Joachim
Ottenhoff, Tom
Guzman, Carlos A
Fletcher, Helen A
Leroy, Odile
Kaufmann, Stefan H E
Helmholtz Centre for Infection Research, Inhoffenstraße 7, 38124 Braunschweig, Germany.
High-throughput analyses of RNA and protein expression are increasingly used for better understanding of vaccine-induced immunity and protection against infectious disease. With an increasing number of vaccine candidates in clinical development, it is timely to consider standardisation and harmonisation of sample collection, storage and analysis to ensure results of highest quality from these precious samples. These challenges were discussed by a group of international experts during a workshop organised by TRANSVAC, a European Commission-funded Research Infrastructure project. The main conclusions were: Platforms are rarely standardised for use in preclinical and clinical studies. Coordinated efforts should continue to harmonise the experimental set up of these studies, as well as the establishment of internal standards and controls. This will ensure comparability, efficiency and feasibility of the global analyses performed on preclinical and clinical data sets.
2015-06-30T13:40:51Z
2015-06-30T13:40:51Z
2014-07-31
Article
TRANSVAC workshop on standardisation and harmonisation of analytical platforms for HIV, TB and malaria vaccines: 'how can big data help?'. 2014, 32 (35):4365-8 Vaccine
1873-2518
24950356
10.1016/j.vaccine.2014.06.014
http://hdl.handle.net/10033/558698
Vaccine
en
info:eu-repo/grantAgreement/EC/FP7/241745
openAccess
oai:repository.helmholtz-hzi.de:10033/6207522018-06-13T15:29:29Zcom_10033_620652col_10033_620666
2017-01-27T08:26:34Z
urn:hdl:10033/620752
The missing link: Bordetella petrii is endowed with both the metabolic versatility of environmental bacteria and virulence traits of pathogenic Bordetellae
Gross, Roy
Guzman, Carlos A
Sebaihia, Mohammed
Martins dos Santos, Vítor A
Pieper, Dietmar H
Koebnik, Ralf
Lechner, Melanie
Bartels, Daniela
Buhrmester, Jens
Choudhuri, Jomuna V
Ebensen, Thomas
Gaigalat, Lars
Herrmann, Stefanie
Khachane, Amit N
Larisch, Christof
Link, Stefanie
Linke, Burkhard
Meyer, Folker
Mormann, Sascha
Nakunst, Diana
Rückert, Christian
Schneiker-Bekel, Susanne
Schulze, Kai
Vorhölter, Frank-Jörg
Yevsa, Tetyana
Engle, Jacquelyn T
Goldman, William E
Pühler, Alfred
Göbel, Ulf B
Goesmann, Alexander
Blöcker, Helmut
Kaiser, Olaf
Martinez-Arias, Rosa
Abstract Background Bordetella petrii is the only environmental species hitherto found among the otherwise host-restricted and pathogenic members of the genus Bordetella. Phylogenetically, it connects the pathogenic Bordetellae and environmental bacteria of the genera Achromobacter and Alcaligenes, which are opportunistic pathogens. B. petrii strains have been isolated from very different environmental niches, including river sediment, polluted soil, marine sponges and a grass root. Recently, clinical isolates associated with bone degenerative disease or cystic fibrosis have also been described. Results In this manuscript we present the results of the analysis of the completely annotated genome sequence of the B. petrii strain DSMZ12804. B. petrii has a mosaic genome of 5,287,950 bp harboring numerous mobile genetic elements, including seven large genomic islands. Four of them are highly related to the clc element of Pseudomonas knackmussii B13, which encodes genes involved in the degradation of aromatics. Though being an environmental isolate, the sequenced B. petrii strain also encodes proteins related to virulence factors of the pathogenic Bordetellae, including the filamentous hemagglutinin, which is a major colonization factor of B. pertussis, and the master virulence regulator BvgAS. However, it lacks all known toxins of the pathogenic Bordetellae. Conclusion The genomic analysis suggests that B. petrii represents an evolutionary link between free-living environmental bacteria and the host-restricted obligate pathogenic Bordetellae. Its remarkable metabolic versatility may enable B. petrii to thrive in very different ecological niches.
2017-01-27T08:26:34Z
2017-01-27T08:26:34Z
2008-09-30
2015-09-04T08:29:04Z
Journal Article
BMC Genomics. 2008 Sep 30;9(1):449
http://dx.doi.org/10.1186/1471-2164-9-449
http://hdl.handle.net/10033/620752
en
Gross et al.
oai:repository.helmholtz-hzi.de:10033/5796032019-08-30T11:28:24Zcom_10033_620652col_10033_620666
2015-10-13T14:30:44Z
urn:hdl:10033/579603
Mitochondrial Ca²⁺ and membrane potential, an alternative pathway for Interleukin 6 to regulate CD4 cell effector function.
Yang, Rui
Lirussi, Dario
Thornton, Tina M
Jelley-Gibbs, Dawn M
Diehl, Sean A
Case, Laure K
Madesh, Muniswamy
Taatjes, Douglas J
Teuscher, Cory
Haynes, Laura
Rincón, Mercedes
Helmholtz Centre for infection research, Inhoffenstr. 7, D-38124 Braunschweig, Germany.
IL-6 plays an important role in determining the fate of effector CD4 cells and the cytokines that these cells produce. Here we identify a novel molecular mechanism by which IL-6 regulates CD4 cell effector function. We show that IL-6-dependent signal facilitates the formation of mitochondrial respiratory chain supercomplexes to sustain high mitochondrial membrane potential late during activation of CD4 cells. Mitochondrial hyperpolarization caused by IL-6 is uncoupled from the production of ATP by oxidative phosphorylation. However, it is a mechanism to raise the levels of mitochondrial Ca(2+) late during activation of CD4 cells. Increased levels of mitochondrial Ca(2+) in the presence of IL-6 are used to prolong Il4 and Il21 expression in effector CD4 cells. Thus, the effect of IL-6 on mitochondrial membrane potential and mitochondrial Ca(2+) is an alternative pathway by which IL-6 regulates effector function of CD4 cells and it could contribute to the pathogenesis of inflammatory diseases.
2015-10-13T14:30:44Z
2015-10-13T14:30:44Z
2015
Article
Mitochondrial Ca²⁺ and membrane potential, an alternative pathway for Interleukin 6 to regulate CD4 cell effector function. 2015, 4: Elife
2050-084X
25974216
10.7554/eLife.06376
http://hdl.handle.net/10033/579603
eLife
en
oai:repository.helmholtz-hzi.de:10033/5796262019-08-30T11:28:51Zcom_10033_620652col_10033_620666
2015-10-14T13:02:28Z
urn:hdl:10033/579626
Rodents as pre-clinical models for predicting vaccine performance in humans.
Riese, Peggy
Trittel, Stephanie
Schulze, Kai
Guzmán, Carlos A
Vaccines represent a key building block for establishing a successful and sustainable control strategy against infectious diseases. Vaccine development often depends on the availability of correlates for protection and reliable animal models for the screening, selection and prioritization of potential vaccine candidates. This is performed according to their immunogenicity, efficacy and safety profiles in pre-clinical studies, which are also critical for identification of candidate antigens, selection of an optimal delivery system and design of appropriate vaccine formulations. Thus, pre-clinical studies in animal models are a prerequisite for addressing crucial issues and generating a solid pre-clinical package for the approval of clinical trials. This review addresses the strengths, limitations and perspectives of rodents as a vaccine development and pre-clinical validation tool.
2015-10-14T13:02:28Z
2015-10-14T13:02:28Z
2015-09
Article
Rodents as pre-clinical models for predicting vaccine performance in humans. 2015, 14 (9):1213-25 Expert Rev Vaccines
1744-8395
26268433
http://hdl.handle.net/10033/579626
Expert review of vaccines
en
oai:repository.helmholtz-hzi.de:10033/5992492019-08-30T11:37:00Zcom_10033_620652col_10033_620666
2016-02-26T10:25:00Z
urn:hdl:10033/599249
Cyclic di-nucleotides: new era for small molecules as adjuvants.
Libanova, Rimma
Becker, Pablo D
Guzmán, Carlos A
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
The implementation of vaccination as an empiric strategy to protect against infectious diseases was introduced even before the advent of hygiene and antimicrobials in the medical practice. Nevertheless, it was not until a few decades ago that we really started understanding the underlying mechanisms of protection triggered by vaccination. Vaccines were initially based on attenuated or inactivated organisms. Subunit vaccines were then introduced as more refined formulations, exhibiting improved safety profiles. However, purified antigens tend to be poorly immunogenic and often require the use of adjuvants to achieve adequate stimulation of the immune system. Vaccination strategies, such as mucosal administration, also require potent adjuvants to improve performance. In the 1990s, immunologists found that pathogens could be sensed as 'danger signals' by receptors recognizing conserved motifs. Although our knowledge is still limited, tremendous advances were made in the understanding of host defence mechanisms regulated by these evolutionary conserved receptors, and the molecular structures which are recognized by them. This opened a new era in adjuvant development. Some of the latest players arrived to this field are the cyclic di-nucleotides, which are ubiquitous prokaryotic intracellular signalling molecules. This review is focused on their potential for the development of vaccines and immunotherapies.
2016-02-26T10:25:00Z
2016-02-26T10:25:00Z
2012-03
Article
Cyclic di-nucleotides: new era for small molecules as adjuvants. 2012, 5 (2):168-76 Microb Biotechnol
1751-7915
21958423
10.1111/j.1751-7915.2011.00306.x
http://hdl.handle.net/10033/599249
Microbial biotechnology
en
info:eu-repo/grantAgreement/EC/FP7/228403
openAccess
oai:repository.helmholtz-hzi.de:10033/6154672019-08-30T11:26:42Zcom_10033_620652col_10033_620666
2016-07-04T14:38:42Z
urn:hdl:10033/615467
Generation and analysis of the improved human HAL9/10 antibody phage display libraries.
Kügler, Jonas
Wilke, Sonja
Meier, Doris
Tomszak, Florian
Frenzel, André
Schirrmann, Thomas
Dübel, Stefan
Garritsen, Henk
Hock, Björn
Toleikis, Lars
Schütte, Mark
Hust, Michael
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Antibody phage display is a proven key technology that allows the generation of human antibodies for diagnostics and therapy. From naive antibody gene libraries - in theory - antibodies against any target can be selected. Here we describe the design, construction and characterization of an optimized antibody phage display library.
2016-07-04T14:38:42Z
2016-07-04T14:38:42Z
2015
Article
Generation and analysis of the improved human HAL9/10 antibody phage display libraries. 2015, 15:10 BMC Biotechnol.
1472-6750
25888378
10.1186/s12896-015-0125-0
http://hdl.handle.net/10033/615467
BMC biotechnology
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6169922019-08-30T11:27:16Zcom_10033_620652col_10033_620666
2016-07-15T11:06:46Z
urn:hdl:10033/616992
The proneurotrophin receptor sortilin is required for Mycobacterium tuberculosis control by macrophages.
Vázquez, Cristina L
Rodgers, Angela
Herbst, Susanne
Coade, Stephen
Gronow, Achim
Guzman, Carlos A
Wilson, Mark S
Kanzaki, Makoto
Nykjaer, Anders
Gutierrez, Maximiliano G
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Sorting of luminal and membrane proteins into phagosomes is critical for the immune function of this organelle. However, little is known about the mechanisms that contribute to the spatiotemporal regulation of this process. Here, we investigated the role of the proneurotrophin receptor sortilin during phagosome maturation and mycobacterial killing. We show that this receptor is acquired by mycobacteria-containing phagosomes via interactions with the adaptor proteins AP-1 and GGAs. Interestingly, the phagosomal association of sortilin is critical for the delivery of acid sphingomyelinase (ASMase) and required for efficient phagosome maturation. Macrophages from Sort1(-/-) mice are less efficient in restricting the growth of Mycobacterium bovis BCG and M. tuberculosis. In vivo, Sort1(-/-) mice showed a substantial increase in cellular infiltration of neutrophils in their lungs and higher bacterial burden after infection with M. tuberculosis. Altogether, sortilin defines a pathway required for optimal intracellular mycobacteria control and lung inflammation in vivo.
2016-07-15T11:06:46Z
2016-07-15T11:06:46Z
2016
Article
The proneurotrophin receptor sortilin is required for Mycobacterium tuberculosis control by macrophages. 2016, 6:29332 Sci Rep
2045-2322
27389464
10.1038/srep29332
http://hdl.handle.net/10033/616992
Scientific reports
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6206862019-08-30T11:33:57Zcom_10033_620652col_10033_620666
2017-01-09T15:08:43Z
urn:hdl:10033/620686
Activation of Innate and Adaptive Immunity by a Recombinant Human Cytomegalovirus Strain Expressing an NKG2D Ligand.
Tomić, Adriana
Varanasi, Pavankumar R
Golemac, Mijo
Malić, Suzana
Riese, Peggy
Borst, Eva M
Mischak-Weissinger, Eva
Guzmán, Carlos A
Krmpotić, Astrid
Jonjić, Stipan
Messerle, Martin
Hel,holtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Development of an effective vaccine against human cytomegalovirus (HCMV) is a need of utmost medical importance. Generally, it is believed that a live attenuated vaccine would best provide protective immunity against this tenacious pathogen. Here, we propose a strategy for an HCMV vaccine that aims at the simultaneous activation of innate and adaptive immune responses. An HCMV strain expressing the host ligand ULBP2 for the NKG2D receptor was found to be susceptible to control by natural killer (NK) cells, and preserved the ability to stimulate HCMV-specific T cells. Infection with the ULBP2-expressing HCMV strain caused diminished cell surface levels of MHC class I molecules. While expression of the NKG2D ligand increased the cytolytic activity of NK cells, NKG2D engagement in CD8+ T cells provided co-stimulation and compensated for lower MHC class I expression. Altogether, our data indicate that triggering of both arms of the immune system is a promising approach applicable to the generation of a live attenuated HCMV vaccine.
2017-01-09T15:08:43Z
2017-01-09T15:08:43Z
2016-12
Article
Activation of Innate and Adaptive Immunity by a Recombinant Human Cytomegalovirus Strain Expressing an NKG2D Ligand. 2016, 12 (12):e1006015 PLoS Pathog.
1553-7374
27907183
10.1371/journal.ppat.1006015
http://hdl.handle.net/10033/620686
PLoS pathogens
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6207422019-08-30T11:36:05Zcom_10033_620652col_10033_620666
2017-01-24T14:40:32Z
urn:hdl:10033/620742
Virus-like particle production with yeast: ultrastructural and immunocytochemical insights into Pichia pastoris producing high levels of the hepatitis B surface antigen.
Lünsdorf, Heinrich
Gurramkonda, Chandrasekhar
Adnan, Ahmad
Khanna, Navin
Rinas, Ursula
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
A protective immune response against Hepatitis B infection can be obtained through the administration of a single viral polypeptide, the Hepatitis B surface antigen (HBsAg). Thus, the Hepatitis B vaccine is generated through the utilization of recombinant DNA technology, preferentially by using yeast-based expression systems. However, the polypeptide needs to assemble into spherical particles, so-called virus-like particles (VLPs), to elicit the required protective immune response. So far, no clear evidence has been presented showing whether HBsAg assembles in vivo inside the yeast cell into VLPs or later in vitro during down-stream processing and purification.
2017-01-24T14:40:32Z
2017-01-24T14:40:32Z
2011-06-26
Article
Virus-like particle production with yeast: ultrastructural and immunocytochemical insights into Pichia pastoris producing high levels of the hepatitis B surface antigen. 2011, 10:48 Microb. Cell Fact.
1475-2859
21703024
10.1186/1475-2859-10-48
http://hdl.handle.net/10033/620742
Microbial cell factories
en
oai:repository.helmholtz-hzi.de:10033/6208172019-08-30T11:29:17Zcom_10033_620652col_10033_620666
2017-02-15T12:06:41Z
urn:hdl:10033/620817
Immune modulator adamantylamide dipeptide stimulates efficient major histocompatibility complex class I-restricted responses in mice.
Becker, Pablo D
Nörder, Miriam
Guzmán, Carlos Alberto
Grinstein, Saul
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Adamantylamide L-alanyl-D-isoglutamine (AdDP) is a synthetic adjuvant which belongs to the family of the desmuramyl peptides. AdDP exerts its adjuvant properties when it is administered either by the parenteral or by the mucosal route, leading to the elicitation of strong humoral responses at both the systemic and the mucosal levels. However, very little is known about the effect of AdDP on cellular immunity. Here we demonstrate that AdDP is able to stimulate cellular responses, which are characterized by the release of gamma interferon by CD8+ T cells when they are restimulated with a major histocompatibility complex class I-restricted peptide and strong in vivo lymphocyte-mediated cytotoxic activity. The capacity of AdDP to stimulate the elicitation of both cellular and humoral adaptive responses makes this adjuvant a promising tool for the development of mucosal vaccine formulations.
2017-02-15T12:06:41Z
2017-02-15T12:06:41Z
2007-05
Article
Immune modulator adamantylamide dipeptide stimulates efficient major histocompatibility complex class I-restricted responses in mice. 2007, 14 (5):538-43 Clin. Vaccine Immunol.
1556-6811
17344349
10.1128/CVI.00316-06
http://hdl.handle.net/10033/620817
Clinical and vaccine immunology : CVI
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6208202019-08-30T11:34:22Zcom_10033_620652col_10033_620666
2017-02-15T13:00:13Z
urn:hdl:10033/620820
Exploitation of prokaryotic expression systems based on the salicylate-dependent control circuit encompassing nahR/P(sal)::xylS2 for biotechnological applications.
Becker, Pablo D
Royo, Jose L
Guzman, Carlos A
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Expression vectors appear to be an indispensable tool for both biological studies and biotechnological applications. Controlling gene overexpression becomes a critical issue when protein production is desired. In addition to several aspects regarding toxicity or plasmid instability, tight control of gene expression is an essential factor in biotechnological processes. Thus, the search for better-controlled circuits is an important issue among biotechnologists. Traditionally, expression systems involve a single regulatory protein operating over a target promoter. However, these circuits are limited on their induction ratios (e.g., by their restriction in the maximal expression capacity, by their leakiness under non-induced conditions). Due to these limitations, regulatory cascades, which are far more efficient, are necessary for biotechnological applications. Thus, regulatory circuits with two modules operating in cascade offer a significant advantage. In this review, we describe the regulatory cascade based on two salicylate-responsive transcriptional regulators of Pseudomonas putida (nahR/P(sal)::xylS2), its properties, and contribution to a tighter control over heterologous gene expression in different applications.Nowadays, heterologous expression has been proven to be an indispensable tool for tackling basic biological questions, as well as for developing biotechnological applications. As the nature of the protein of interest becomes more complex, biotechnologists find that a tight control of gene expression is a key factor which conditions the success of the downstream purification process, as well as the interpretation of the results in other type of studies. Fortunately, different expression systems can be found in the market, each of them with their own pros and cons. In this review we discuss the exploitation of prokaryotic expression systems based on a promising expression system, the salicylate-dependent control circuit encompassing nahR/P(sal)::xylS2, as well as some of the improvements that have been done on this system to exploit it more efficiently in the context of both biotechnological applications and basic research.
2017-02-15T13:00:13Z
2017-02-15T13:00:13Z
2017-02-15
Article
Exploitation of prokaryotic expression systems based on the salicylate-dependent control circuit encompassing nahR/P(sal)::xylS2 for biotechnological applications., 1 (4):244-51 Bioeng Bugs
1949-1026
21327056
10.4161/bbug.1.4.11247
http://hdl.handle.net/10033/620820
Bioengineered bugs
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6208532019-08-30T11:29:17Zcom_10033_620652col_10033_620666
2017-03-08T14:36:40Z
urn:hdl:10033/620853
Whole-Genome Enrichment Provides Deep Insights into Vibrio cholerae Metagenome from an African River.
Vezzulli, L
Grande, C
Tassistro, G
Brettar, I
Höfle, M G
Pereira, R P A
Mushi, D
Pallavicini, A
Vassallo, P
Pruzzo, C
Helmholtz Centre for infection research, Ihoffenstr. 7, 38124 Braunschweig, Germany.
The detection and typing of Vibrio cholerae in natural aquatic environments encounter major methodological challenges related to the fact that the bacterium is often present in environmental matrices at very low abundance in nonculturable state. This study applied, for the first time to our knowledge, a whole-genome enrichment (WGE) and next-generation sequencing (NGS) approach for direct genotyping and metagenomic analysis of low abundant V. cholerae DNA (<50 genome unit/L) from natural water collected in the Morogoro river (Tanzania). The protocol is based on the use of biotinylated RNA baits for target enrichment of V. cholerae metagenomic DNA via hybridization. An enriched V. cholerae metagenome library was generated and sequenced on an Illumina MiSeq platform. Up to 1.8 × 10(7) bp (4.5× mean read depth) were found to map against V. cholerae reference genome sequences representing an increase of about 2500 times in target DNA coverage compared to theoretical calculations of performance for shotgun metagenomics. Analysis of metagenomic data revealed the presence of several V. cholerae virulence and virulence associated genes in river water including major virulence regions (e.g. CTX prophage and Vibrio pathogenicity island-1) and genetic markers of epidemic strains (e.g. O1-antigen biosynthesis gene cluster) that were not detectable by standard culture and molecular techniques. Overall, besides providing a powerful tool for direct genotyping of V. cholerae in complex environmental matrices, this study provides a 'proof of concept' on the methodological gap that might currently preclude a more comprehensive understanding of toxigenic V. cholerae emergence from natural aquatic environments.
2017-03-08T14:36:40Z
2017-03-08T14:36:40Z
2016-11-25
Article
Whole-Genome Enrichment Provides Deep Insights into Vibrio cholerae Metagenome from an African River. 2016: Microb. Ecol.
1432-184X
27888291
10.1007/s00248-016-0902-x
http://hdl.handle.net/10033/620853
Microbial ecology
en
info:eu-repo/grantAgreement/EC/FP7/311846
http://creativecommons.org/licenses/by-nc-sa/4.0/
openAccess
oai:repository.helmholtz-hzi.de:10033/6208702019-08-30T11:29:47Zcom_10033_620652col_10033_620666
2017-03-24T10:49:16Z
urn:hdl:10033/620870
Immunization with Tc52 or its amino terminal domain adjuvanted with c-di-AMP induces Th17+Th1 specific immune responses and confers protection against Trypanosoma cruzi.
Matos, Marina N
Cazorla, Silvia I
Schulze, Kai
Ebensen, Thomas
Guzmán, Carlos Alberto
Malchiodi, Emilio L
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
The development of new adjuvants enables fine modulation of the elicited immune responses. Ideally, the use of one or more adjuvants should result in the induction of a protective immune response against the specific pathogen. We have evaluated the immune response and protection against Trypanosoma cruzi infection in mice vaccinated with recombinant Tc52 or its N- and C-terminal domains (NTc52 and CTc52) adjuvanted either with the STING (Stimulator of Interferon Genes) agonist cyclic di-AMP (c-di-AMP), a pegylated derivative of α-galactosylceramide (αGC-PEG), or oligodeoxynucleotides containing unmethylated CpG motifs (ODN-CpG). All groups immunized with the recombinant proteins plus adjuvant: Tc52+c-di-AMP, NTc52+c-di-AMP, CTc52+c-di-AMP, NTc52+c-di-AMP+αGC-PEG, NTc52+CpG, developed significantly higher anti-Tc52 IgG titers than controls. Groups immunized with c-di-AMP and Tc52, NTc52 or CTc52 showed the highest Tc52-specific IgA titers in nasal lavages. All groups immunized with the recombinant proteins plus adjuvant developed a strong specific cellular immune response in splenocytes and lymph node cells with significant differences for groups immunized with c-di-AMP and Tc52, NTc52 or CTc52. These groups also showed high levels of Tc52-specific IL-17 and IFN-γ producing cells, while NTc52+CpG group only showed significant difference with control in IFN-γ producing cells. Groups immunized with c-di-AMP and Tc52, NTc52 or CTc52 developed predominantly a Th17 and Th1immune response, whereas for NTc52+CpG it was a dominant Th1 response. It was previously described that αGC-PEG inhibits Th17 differentiation by activating NKT cells. Thus, in this work we have also included a group immunized with both adjuvants (NTc52+c-di-AMP+αGC-PEG) with the aim to modulate the Th17 response induced by c-di-AMP. This group showed a significant reduction in the number of Tc52-specific IL-17 producing splenocytes, as compared to the group NTc52+c-di-AMP, which has in turn correlated with a reduction in protection against infection. These results suggest that the Th17 immune response developed after immunizing with NTc52+c-di-AMP could have a protective role against T. cruzi infection. Groups NTc52+c-di-AMP, Tc52+c-di-AMP and NTc52PB, were the ones that showed better protection against infection with lower parasitemia and weight loss, and higher survival.
2017-03-24T10:49:16Z
2017-03-24T10:49:16Z
2017-02
Article
Immunization with Tc52 or its amino terminal domain adjuvanted with c-di-AMP induces Th17+Th1 specific immune responses and confers protection against Trypanosoma cruzi. 2017, 11 (2):e0005300 PLoS Negl Trop Dis
1935-2735
28234897
10.1371/journal.pntd.0005300
http://hdl.handle.net/10033/620870
PLoS neglected tropical diseases
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6209212019-08-30T11:29:47Zcom_10033_620652col_10033_620666
2017-05-17T12:07:22Z
urn:hdl:10033/620921
Blockade of Neutrophil's Chemokine Receptors CXCR1/2 Abrogate Liver Damage in Acute-on-Chronic Liver Failure.
Khanam, Arshi
Trehanpati, Nirupma
Riese, Peggy
Rastogi, Archana
Guzman, Carlos A.
Sarin, Shiv Kumar
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Neutrophils serve as critical players in the pathogenesis of liver diseases. Chemokine receptors CXCR1 and CXCR2 are required for neutrophil chemotaxis to the site of inflammation/injury and are crucial in hepatic inflammatory response. However, key mechanism of neutrophil-mediated liver injury in acute-on-chronic liver failure (ACLF) remains highly elusive; which could be targeted for the development of new therapeutic interventions.
2017-05-17T12:07:22Z
2017-05-17T12:07:22Z
2017
Article
Blockade of Neutrophil's Chemokine Receptors CXCR1/2 Abrogate Liver Damage in Acute-on-Chronic Liver Failure. 2017, 8:464 Front Immunol
28484461
10.3389/fimmu.2017.00464
http://hdl.handle.net/10033/620921
Frontiers in immunology
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6209832019-08-30T11:30:32Zcom_10033_620652col_10033_620666
2017-06-28T08:17:43Z
urn:hdl:10033/620983
Gene Expression Driven by a Strong Viral Promoter in MVA Increases Vaccination Efficiency by Enhancing Antibody Responses and Unmasking CD8⁺ T Cell Epitopes.
Becker, Pablo D
Nörder, Miriam
Weissmann, Sebastian
Ljapoci, Ronny
Erfle, Volker
Drexler, Ingo
Guzmán, Carlos Alberto
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Viral vectors are promising tools for vaccination strategies and immunotherapies. However, CD8⁺ T cell responses against pathogen-derived epitopes are usually limited to dominant epitopes and antibody responses to recombinant encoded antigens (Ags) are mostly weak. We have previously demonstrated that the timing of viral Ag expression in infected professional Ag-presenting cells strongly shapes the epitope immunodominance hierarchy. T cells recognizing determinants derived from late viral proteins have a clear disadvantage to proliferate during secondary responses. In this work we evaluate the effect of overexpressing the recombinant Ag using the modified vaccinia virus early/late promoter H5 (mPH5). Although the Ag-expression from the natural promoter 7.5 (P7.5) and the mPH5 seemed similar, detailed analysis showed that mPH5 not only induces higher expression levels than P7.5 during early phase of infection, but also Ag turnover is enhanced. The strong overexpression during the early phase leads to broader CD8 T cell responses, while preserving the priming efficiency of stable Ags. Moreover, the increase in Ag-secretion favors the induction of strong antibody responses. Our findings provide the rationale to develop new strategies for fine-tuning the responses elicited by recombinant modified vaccinia virus Ankara by using selected promoters to improve the performance of this viral vector.
2017-06-28T08:17:43Z
2017-06-28T08:17:43Z
2014-07-22
Article
Gene Expression Driven by a Strong Viral Promoter in MVA Increases Vaccination Efficiency by Enhancing Antibody Responses and Unmasking CD8⁺ T Cell Epitopes. 2014, 2 (3):581-600 Vaccines (Basel)
2076-393X
26344747
10.3390/vaccines2030581
http://hdl.handle.net/10033/620983
Vaccines
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6210132019-08-30T11:26:13Zcom_10033_211390com_10033_620601com_10033_620652col_10033_620666col_10033_211409col_10033_620603
2017-07-14T13:15:38Z
urn:hdl:10033/621013
Anti-nuclear autoantibodies in the general German population: prevalence and lack of association with selected cardiovascular and metabolic disorders-findings of a multicenter population-based study.
Akmatov, Manas K
Röber, Nadja
Ahrens, Wolfgang
Flesch-Janys, Dieter
Fricke, Julia
Greiser, Halina
Günther, Kathrin
Kaaks, Rudolf
Kemmling, Yvonne
Krone, Bastian
Linseisen, Jakob
Meisinger, Christa
Moebus, Susanne
Obi, Nadia
Guzman, Carlos A
Conrad, Karsten
Pessler, Frank
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
We determined the prevalence of anti-nuclear autoantibodies (ANAs) in the German adult population and examined the association between ANAs and cardiovascular and metabolic disorders.
2017-07-14T13:15:38Z
2017-07-14T13:15:38Z
2017-06-06
Article
Anti-nuclear autoantibodies in the general German population: prevalence and lack of association with selected cardiovascular and metabolic disorders-findings of a multicenter population-based study. 2017, 19 (1):127 Arthritis Res. Ther.
1478-6362
28587625
10.1186/s13075-017-1338-5
http://hdl.handle.net/10033/621013
Arthritis research & therapy
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6210372019-08-30T11:31:23Zcom_10033_620601com_10033_620652col_10033_620666col_10033_620602
2017-08-03T13:05:38Z
urn:hdl:10033/621037
Type I IFN and not TNF, is Essential for Cyclic Di-nucleotide-elicited CTL by a Cytosolic Cross-presentation Pathway.
Lirussi, Darío
Ebensen, Thomas
Schulze, Kai
Trittel, Stephanie
Duran, Veronica
Liebich, Ines
Kalinke, Ulrich
Guzmán, Carlos Alberto
Helmholtz Centre for infection research, Inhoffenstr.7, 38124 Braunschweig, Germany.
Cyclic di-nucleotides (CDN) are potent stimulators of innate and adaptive immune responses. Cyclic di-AMP (CDA) is a promising adjuvant that generates humoral and cellular immunity. The strong STING-dependent stimulation of type I IFN represents a key feature of CDA. However, recent studies suggested that this is dispensable for adjuvanticity. Here we demonstrate that stimulation of IFN-γ-secreting CD8(+) cytotoxic T lymphocytes (CTL) is significantly decreased after vaccination in the absence of type I IFN signaling. The biological significance of this CTL response was confirmed by the stimulation of MHC class I-restricted protection against influenza virus challenge. We show here that type I IFN (and not TNF-α) is essential for CDA-mediated cross-presentation by a cathepsin independent, TAP and proteosome dependent cytosolic antigen processing pathway, which promotes effective cross-priming and further CTL induction. Our data clearly demonstrate that type I IFN signaling is critical for CDN-mediated cross-presentation.
2017-08-03T13:05:38Z
2017-08-03T13:05:38Z
2017-07-19
Article
Type I IFN and not TNF, is Essential for Cyclic Di-nucleotide-elicited CTL by a Cytosolic Cross-presentation Pathway. 2017 EBioMedicine
2352-3964
28754303
10.1016/j.ebiom.2017.07.016
http://hdl.handle.net/10033/621037
EBioMedicine
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6210672019-08-30T11:27:16Zcom_10033_620652col_10033_620666
2017-08-21T14:05:36Z
urn:hdl:10033/621067
Cell aggregation enhances bone formation by human mesenchymal stromal cells.
Chatterjea, A
LaPointe, V L
Barradas, A
Garritsen, H
Yuan, H
Renard, A
van Blitterswijk, C A
de Beor, J
Hemholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Vraunschweig, Germany.
The amount of bone generated using current tissue engineering approaches is insufficient for many clinical applications. Previous in vitro studies suggest that culturing cells as 3D aggregates can enhance their osteogenic potential, but the effect on bone formation in vivo is unknown. Here, we use agarose wells to generate uniformly sized mesenchymal stromal cell (MSC) aggregates. When combined with calcium phosphate ceramic particles and a gel prepared from human platelet-rich plasma, we generated a tissue engineered construct which significantly improved in vivo bone forming capacity as compared to the conventional system of using single cells seeded directly on the ceramic surface. Histology demonstrated the reproducibility of this system, which was tested using cells from four different donors. In vitro studies established that MSC aggregation results in an up-regulation of osteogenic transcripts. And finally, the in vivo performance of the constructs was significantly diminished when unaggregated cells were used, indicating that cell aggregation is a potent trigger of in vivo bone formation by MSCs. Cell aggregation could thus be used to improve bone tissue engineering strategies.
2017-08-21T14:05:36Z
2017-08-21T14:05:36Z
2017-02-15
Article
Cell aggregation enhances bone formation by human mesenchymal stromal cells. 2017, 33:121-129 Eur Cell Mater
1473-2262
28198985
10.22203/eCM.v033a09
http://hdl.handle.net/10033/621067
European cells & materials
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6211052019-08-30T11:35:14Zcom_10033_620652col_10033_620666
2017-09-12T10:27:10Z
urn:hdl:10033/621105
Intranasal vaccination with an adjuvanted polyphosphazenes nanoparticle-based vaccine formulation stimulates protective immune responses in mice.
Schulze, Kai
Ebensen, Thomas
Babiuk, Lorne A
Gerdts, Volker
Guzman, Carlos A.
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr.7, 38124 Braunschweig, Germany.
The most promising strategy to sustainably prevent infectious diseases is vaccination. However, emerging as well as re-emerging diseases still constitute a considerable threat. Furthermore, lack of compliance and logistic constrains often result in the failure of vaccination campaigns. To overcome these hurdles, novel vaccination strategies need to be developed, which fulfill maximal safety requirements, show maximal efficiency and are easy to administer. Mucosal vaccines constitute promising non-invasive approaches able to match these demands. Here we demonstrate that nanoparticle (polyphosphazenes)-based vaccine formulations including c-di-AMP as adjuvant, cationic innate defense regulator peptides (IDR) and ovalbumin (OVA) as model antigen were able to stimulate strong humoral and cellular immune responses, which conferred protection against the OVA expressing influenza strain A/WSN/OVAI (H1N1). The presented results confirm the potency of nanoparticle-based vaccine formulations to deliver antigens across the mucosal barrier, but also demonstrate the necessity to include adjuvants to stimulate efficient antigen-specific immune responses.
2017-09-12T10:27:10Z
2017-09-12T10:27:10Z
2017-06-01
Article
Intranasal vaccination with an adjuvanted polyphosphazenes nanoparticle-based vaccine formulation stimulates protective immune responses in mice. 2017, 13 (7):2169-2178 Nanomedicine
1549-9642
28579436
10.1016/j.nano.2017.05.012
http://hdl.handle.net/10033/621105
Nanomedicine : nanotechnology, biology, and medicine
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6211242019-08-30T11:27:16Zcom_10033_620652col_10033_620666
2017-09-27T08:58:10Z
urn:hdl:10033/621124
Bivalent mucosal peptide vaccines administered using the LCP carrier system stimulate protective immune responses against Streptococcus pyogenes infection.
Schulze, Kai
Ebensen, Thomas
Chandrudu, Saranya
Skwarczynski, Mariusz
Toth, Istvan
Olive, Colleen
Guzman, Carlos A
Helmholtz Centre for infection researchGmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Despite the broad knowledge about the pathogenicity of Streptococcus pyogenes there is still a controversy about the correlate of protection in GAS infections. We aimed in further improving the immune responses stimulated against GAS comparing different vaccine formulations including bis-(3',5')-cyclic dimeric adenosine monophosphate (c-di-AMP) and BPPCysMPEG, a derivative of the macrophage-activating lipopeptide (MALP-2), as adjuvants, respectively, to be administered with and without the universal T helper cell epitope P25 along with the optimized B cell epitope J14 of the M protein and B and T cell epitopes of SfbI. Lipopeptide based nano carrier systems (LCP) were used for efficient antigen delivery across the mucosal barrier. The stimulated immune responses were efficient in protecting mice against a respiratory challenge with a lethal dose of a heterologous S. pyogenes strain. Moreover, combination of the LCP based peptide vaccine with c-di-AMP allowed reduction of antigen dose at the same time maintaining vaccine efficacy.
2017-09-27T08:58:10Z
2017-09-27T08:58:10Z
2017-09-05
Article
Bivalent mucosal peptide vaccines administered using the LCP carrier system stimulate protective immune responses against Streptococcus pyogenes infection. 2017, 13 (8):2463-2474 Nanomedicine
1549-9642
28887213
10.1016/j.nano.2017.08.015
http://hdl.handle.net/10033/621124
Nanomedicine : nanotechnology, biology, and medicine
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6211272019-08-30T11:30:58Zcom_10033_620652col_10033_620666
2017-10-05T11:16:20Z
urn:hdl:10033/621127
Roads to advanced vaccines: influenza case study.
Riese, Peggy
Guzmán, Carlos A
Helmholz Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Vaccines represent a cornerstone to ensure healthy lives and promote well-being for all at all ages. However, there are many diseases for which vaccines are not available, are relatively ineffective or need to be adapted periodically. Advances in microbial biotechnology will contribute to overcoming these roadblocks by laying the groundwork for improving and creating new approaches for developing better vaccines, as illustrated here in the case of influenza.
2017-10-05T11:16:20Z
2017-10-05T11:16:20Z
2017-09
Article
Roads to advanced vaccines: influenza case study. 2017, 10 (5):1036-1040 Microb Biotechnol
1751-7915
28809451
10.1111/1751-7915.12835
http://hdl.handle.net/10033/621127
Microbial biotechnology
en
info:eu-repo/grantAgreement/EC/FP7/601738
http://creativecommons.org/licenses/by-nc-sa/4.0/
openAccess
oai:repository.helmholtz-hzi.de:10033/6211762019-08-30T11:26:42Zcom_10033_620652col_10033_620666
2017-11-15T13:53:29Z
urn:hdl:10033/621176
Mucosal Administration of Cycle-Di-Nucleotide-Adjuvanted Virosomes Efficiently Induces Protection against Influenza H5N1 in Mice.
Ebensen, Thomas
Debarry, Jennifer
Pedersen, Gabriel K
Blazejewska, Paulina
Weissmann, Sebastian
Schulze, Kai
McCullough, Kenneth C
Cox, Rebecca J
Guzmán, Carlos A
Helmholtz Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
The need for more effective influenza vaccines is highlighted by the emergence of novel influenza strains, which can lead to new pandemics. There is a growing population of susceptible subjects at risk for severe complications of influenza, such as the elderly who are only in part protected by current licensed seasonal vaccines. One strategy for improving seasonal and pandemic vaccines takes advantage of adjuvants to boost and modulate evoked immune responses. In this study, we examined the capacity of the recently described adjuvant cyclic di-adenosine monophosphate (c-di-AMP) to serve as an adjuvant for improved mucosal influenza vaccines, and induce effective protection against influenza H5N1. In detail, c-di-AMP promoted (i) effective local and systemic humoral immune responses, including protective hemagglutination inhibition titers, (ii) effective cellular responses, including multifunctional T cell activity, (iii) induction of long-lasting immunity, and (iv) protection against viral challenge. Furthermore, we demonstrated the dose-sparing capacity of the adjuvant as well as the ability to evoke cross-clade protective immune responses. Overall, our results suggest that c-di-AMP contributes to the generation of a protective cell-mediated immune response required for efficacious vaccination against influenza, which supports the further development of c-di-AMP as an adjuvant for seasonal and pandemic influenza mucosal vaccines.
2017-11-15T13:53:29Z
2017-11-15T13:53:29Z
2017
Article
Mucosal Administration of Cycle-Di-Nucleotide-Adjuvanted Virosomes Efficiently Induces Protection against Influenza H5N1 in Mice. 2017, 8:1223 Front Immunol
1664-3224
29033942
10.3389/fimmu.2017.01223
http://hdl.handle.net/10033/621176
Frontiers in immunology
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6211942019-08-30T11:25:11Zcom_10033_620652col_10033_620666
2017-12-05T09:01:58Z
urn:hdl:10033/621194
Superior immunogenicity of HCV envelope glycoproteins when adjuvanted with cyclic-di-AMP, a STING activator or archaeosomes.
Landi, A
Law, J
Hockman, D
Logan, M
Crawford, K
Chen, C
Kundu, J
Ebensen, T
Guzman, C A
Deschatelets, L
Krishnan, L
Tyrrell, D L J
Houghton, M
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr.7, 38124 Braunschweig, Germany.
Three decades after the discovery, hepatitis C virus (HCV) is still the leading cause of liver transplantation and poses a major threat to global health. In spite of recent advances in the development of direct acting antivirals, there is still a need for a prophylactic vaccine to limit the virus spread and protect at-risk populations, especially in developing countries, where the cost of the new treatments may severely limit access. The use of recombinant HCV glycoproteins E1E2 (rE1E2) in combination with the MF59, an oil-in-water emulsion-based adjuvant, has previously been shown to reduce the rate of chronicity in chimpanzees and to induce production of cross-neutralizing antibodies and cellular immune responses in human volunteers. To further improve neutralizing antibody responses in recipients along with robust T cell responses, we have explored the immunogenicity of different adjuvants when formulated with the HCV rE1E2 vaccine in mice. Our data show that cyclic di-adenosine monophosphate (c-di-AMP) and archaeosomes elicit strong neutralizing antibodies similar to those elicited using aluminum hydroxide/monophosphoryl lipid A (Alum/monophos. /MPLA) and MF59. However, both c-di-AMP and archaeosomes induced a more robust cellular immune response, which was confirmed by the detection of vaccine-specific poly-functional CD4+ T cells. We conclude that these adjuvants may substantially boost the immunogenicity of our E1E2 vaccine. In addition, our data also indicates that use of a partial or exclusive intranasal immunization regimen may also be feasible using c-di-AMP as adjuvant.
2017-12-05T09:01:58Z
2017-12-05T09:01:58Z
2017-12-15
Article
Superior immunogenicity of HCV envelope glycoproteins when adjuvanted with cyclic-di-AMP, a STING activator or archaeosomes. 2017, 35 (50):6949-6956 Vaccine
1873-2518
29089195
10.1016/j.vaccine.2017.10.072
http://hdl.handle.net/10033/621194
Vaccine
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6212162021-07-06T12:05:05Zcom_10033_620652com_10033_620601com_10033_620591com_10033_622921col_10033_620725col_10033_620666col_10033_622926col_10033_620602
2018-01-02T13:05:22Z
urn:hdl:10033/621216
Targeted antigen delivery to dendritic cells elicits robust antiviral T cell-mediated immunity in the liver.
Volckmar, Julia
Gereke, Marcus
Ebensen, Thomas
Riese, Peggy
Philipsen, Lars
Lienenklaus, Stefan
Wohlleber, Dirk
Klopfleisch, Robert
Stegemann-Koniszewski, Sabine
Müller, Andreas J
Gruber, Achim D
Knolle, Percy
Guzman, Carlos A
Bruder, Dunja
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr.7, 38124 Braunschweig, Germany.
Hepatotropic viruses such as hepatitis C virus cause life-threatening chronic liver infections in millions of people worldwide. Targeted in vivo antigen-delivery to cross-presenting dendritic cells (DCs) has proven to be extraordinarily efficient in stimulating antigen-specific T cell responses. To determine whether this approach would as well be suitable to induce local antiviral effector T cells in the liver we compared different vaccine formulations based on either the targeting of DEC-205 or TLR2/6 on cross-presenting DCs or formulations not involving in vivo DC targeting. As read-outs we used in vivo hepatotropic adenovirus challenge, histology and automated multidimensional fluorescence microscopy (MELC). We show that targeted in vivo antigen delivery to cross-presenting DCs is highly effective in inducing antiviral CTLs capable of eliminating virus-infected hepatocytes, while control vaccine formulation not involving DC targeting failed to induce immunity against hepatotropic virus. Moreover, we observed distinct patterns of CD8+ T cell interaction with virus-infected and apoptotic hepatocytes in the two DC-targeting groups suggesting that the different vaccine formulations may stimulate distinct types of effector functions. Our findings represent an important step toward the future development of vaccines against hepatotropic viruses and the treatment of patients with hepatic virus infection after liver transplantation to avoid reinfection.
2018-01-02T13:05:22Z
2018-01-02T13:05:22Z
2017-03-07
Article
Targeted antigen delivery to dendritic cells elicits robust antiviral T cell-mediated immunity in the liver. 2017, 7:43985 Sci Rep
2045-2322
28266658
10.1038/srep43985
http://hdl.handle.net/10033/621216
Scientific reports
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6212682019-08-30T11:26:13Zcom_10033_620626com_10033_620636com_10033_620652col_10033_620666col_10033_620627col_10033_620638
2018-02-08T15:10:19Z
urn:hdl:10033/621268
Deletion of Irf3 and Irf7 Genes in Mice Results in Altered Interferon Pathway Activation and Granulocyte-Dominated Inflammatory Responses to Influenza A Infection.
Hatesuer, Bastian
Hoang, Hang Thi Thu
Riese, Peggy
Trittel, Stephanie
Gerhauser, Ingo
Elbahesh, Husni
Geffers, Robert
Wilk, Esther
Schughart, Klaus
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr.7, 38124 Braunschweig, Germany.
The interferon (IFN) pathway plays an essential role in the innate immune response following viral infections and subsequent shaping of adaptive immunity. Infections with influenza A viruses (IAV) activate the IFN pathway after the recognition of pathogen-specific molecular patterns by respective pattern recognition receptors. The IFN regulatory factors IRF3 and IRF7 are key players in the regulation of type I and III IFN genes. In this study, we analyzed the role of IRF3 and IRF7 for the host response to IAV infections in Irf3-/-, Irf7-/-, and Irf3-/-Irf7-/- knockout mice. While the absence of IRF3 had only a moderate impact on IFN expression, deletion of IRF7 completely abolished IFNα production after infection. In contrast, lack of both IRF3 and IRF7 resulted in the absence of both IFNα and IFNβ after IAV infection. In addition, IAV infection of double knockout mice resulted in a strong increase of mortality associated with a massive influx of granulocytes in the lung and reduced activation of the adaptive immune response.
2018-02-08T15:10:19Z
2018-02-08T15:10:19Z
2017
Article
Deletion of Irf3 and Irf7 Genes in Mice Results in Altered Interferon Pathway Activation and Granulocyte-Dominated Inflammatory Responses to Influenza A Infection. 2017, 9 (2):145-161 J Innate Immun
1662-8128
27811478
10.1159/000450705
http://hdl.handle.net/10033/621268
Journal of innate immunity
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6213592019-08-30T11:33:29Zcom_10033_620652col_10033_620666
2018-04-23T12:28:18Z
urn:hdl:10033/621359
Engineered trivalent immunogen adjuvanted with a STING agonist confers protection against Trypanosoma cruzi infection.
Sanchez Alberti, Andrés
Bivona, Augusto E
Cerny, Natacha
Schulze, Kai
Weißmann, Sebastian
Ebensen, Thomas
Morales, Celina
Padilla, Angel M
Cazorla, Silvia I
Tarleton, Rick L
Guzmán, Carlos A
Malchiodi, Emilio L
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
The parasite Trypanosoma cruzi is the causative agent of Chagas disease, a potentially life-threatening infection that represents a major health problem in Latin America. Several characteristics of this protozoan contribute to the lack of an effective vaccine, among them: its silent invasion mechanism, T. cruzi antigen redundancy and immunodominance without protection. Taking into account these issues, we engineered Traspain, a chimeric antigen tailored to present a multivalent display of domains from key parasitic molecules, combined with stimulation of the STING pathway by c-di-AMP as a novel prophylactic strategy. This formulation proved to be effective for the priming of functional humoral responses and pathogen-specific CD8+ and CD4+ T cells, compatible with a Th1/Th17 bias. Interestingly, vaccine effectiveness assessed across the course of infection, showed a reduction in parasite load and chronic inflammation in different proof of concept assays. In conclusion, this approach represents a promising tool against parasitic chronic infections.
2018-04-23T12:28:18Z
2018-04-23T12:28:18Z
2017
Article
Engineered trivalent immunogen adjuvanted with a STING agonist confers protection against Trypanosoma cruzi infection. 2017, 2:9 NPJ Vaccines
2059-0105
29263868
10.1038/s41541-017-0010-z
http://hdl.handle.net/10033/621359
NPJ vaccines
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
oai:repository.helmholtz-hzi.de:10033/6214152019-08-30T11:25:40Zcom_10033_620652col_10033_620666
2018-06-28T14:40:15Z
urn:hdl:10033/621415
Self-Amplifying Replicon RNA Delivery to Dendritic Cells by Cationic Lipids
Englezou, Pavlos C.
Sapet, Cedric
Démoulins, Thomas
Milona, Panagiota
Ebensen, Thomas
Schulze, Kai
Guzman, Carlos-Alberto
Poulhes, Florent
Zelphati, Olivier
Ruggli, Nicolas
McCullough, Kenneth C.
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
2018-06-28T14:40:15Z
2018-06-28T14:40:15Z
Article
21622531
10.1016/j.omtn.2018.04.019
http://hdl.handle.net/10033/621415
http://linkinghub.elsevier.com/retrieve/pii/S2162253118300970
info:eu-repo/grantAgreement/EC/FP7/251420
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
openAccess
Attribution-NonCommercial-ShareAlike 3.0 United States
12
118
134
Molecular Therapy - Nucleic Acids
oai:repository.helmholtz-hzi.de:10033/6214312020-10-07T01:33:44Zcom_10033_620652com_10033_620636col_10033_620666col_10033_620665
2018-07-25T09:40:30Z
urn:hdl:10033/621431
Induction of CD4(+) and CD8(+) anti-tumor effector T cell responses by bacteria mediated tumor therapy.
Stern, Christian
Kasnitz, Nadine
Kocijancic, Dino
Trittel, Stephanie
Riese, Peggy
Guzman, Carlos A
Leschner, Sara
Weiss, Siegfried
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
CD4+
CD8+
E. coli Top10
T cells
tumor antigen presentation
tumor necrosis
Facultative anaerobic bacteria like E. coli can colonize solid tumors often resulting in tumor growth retardation or even clearance. Little mechanistic knowledge is available for this phenomenon which is however crucial for optimization and further implementation in the clinic. Here, we show that intravenous injections with E. coli TOP10 can induce clearance of CT26 tumors in BALB/c mice. Importantly, re-challenging mice which had cleared tumors showed that clearance was due to a specific immune reaction. Accordingly, lymphopenic mice never showed tumor clearance after infection. Depletion experiments revealed that during induction phase, CD8(+) T cells are the sole effectors responsible for tumor clearance while in the memory phase CD8(+) and CD4(+) T cells were involved. This was confirmed by adoptive transfer. CD4(+) and CD8(+) T cells could reject newly set tumors while CD8(+) T cells could even reject established tumors. Detailed analysis of adoptively transferred CD4(+) T cells during tumor challenge revealed expression of granzyme B, FasL, TNF-α and IFN-γ in such T cells that might be involved in the anti-tumor activity. Our findings should pave the way for further optimization steps of this promising therapy.
2018-07-25T09:40:30Z
2018-07-25T09:40:30Z
2015-10-15
Article
1097-0215
25868911
10.1002/ijc.29567
http://hdl.handle.net/10033/621431
https://onlinelibrary.wiley.com/doi/abs/10.1002/ijc.29567
International journal of cancer
oai:repository.helmholtz-hzi.de:10033/6214372019-08-30T11:34:44Zcom_10033_620652col_10033_620666
2018-08-07T09:25:04Z
urn:hdl:10033/621437
Large-scale production of megakaryocytes in microcarrier-supported stirred suspension bioreactors.
Eicke, Dorothee
Baigger, Anja
Schulze, Kai
Latham, Sharissa L
Halloin, Caroline
Zweigerdt, Robert
Guzman, Carlos A
Blasczyk, Rainer
Figueiredo, Constança
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Megakaryocytes (MKs) are the precursors of platelets (PLTs) and may be used for PLT production in vivo or in vitro, as well as a source for PLT-derived growth factors. Induced pluripotent stem cells represent an unlimited cell source for the in vitro production of MKs. This study aimed at developing an effective, xeno-free and scalable system to produce high numbers of MKs. In particular, microcarrier beads-assisted stirred bioreactors were evaluated as a means of improving MK yields. This method resulted in the production of 18.7 × 10
2018-08-07T09:25:04Z
2018-08-07T09:25:04Z
2018-07-05
Article
2045-2322
29977045
10.1038/s41598-018-28459-x
http://hdl.handle.net/10033/621437
en
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Attribution-NonCommercial-ShareAlike 3.0 United States
Scientific reports
oai:repository.helmholtz-hzi.de:10033/6215712019-08-30T11:29:43Zcom_10033_620652col_10033_620666
2018-11-16T14:40:58Z
urn:hdl:10033/621571
Rapid In Vivo Assessment of Adjuvant's Cytotoxic T Lymphocytes Generation Capabilities for Vaccine Development
Lirussi, Darío
Ebensen, Thomas
Schulze, Kai
Reinhard, Elena
Trittel, Stephanie
Riese, Peggy
Prochnow, Blair
Guzmán, Carlos A.
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
The assessment of modern sub-unit vaccines reveals that the generation of neutralizing antibodies is important but not sufficient for adjuvant selection. Therefore, adjuvants with both humoral and cellular immuno-stimulatory capabilities that are able to promote cytotoxic T lymphocytes (CTL) responses are urgently needed. Thus, faithful monitoring of adjuvant candidates that induce cross-priming and subsequently enhance CTL generation represents a crucial step in vaccine development. In here we present an application for a method that uses SIINFEKL-specific (OT-I) T cells to monitor the cross-presentation of the model antigen ovalbumin (OVA) in vivo in the presence of different adjuvant candidates. This method represents a rapid test to select adjuvants with the best cross-priming capabilities. The proliferation of CD8+ T cells is the most valuable indication of cross-priming and it is also regarded as a correlate of adjuvant-induced cross-presentation. This feature can be evaluated in different immune organs like lymph nodes and spleen. The extent of the CTL generation can also be monitored, thereby giving insights on the nature of a local (draining lymph node mainly) or a systemic response (distant lymph nodes and/or spleen). This technique further allows multiple modifications for testing drugs that can inhibit specific cross-presentation pathways and also offers the possibility to be used in different strains of conventional and genetically modified mice. In summary, the application that we present here will be useful for vaccine laboratories in industry or academia that develop or modify chemical adjuvants for vaccine research and development. © 2018, Journal of Visualized Experiments.
2018-11-16T14:40:58Z
2018-11-16T14:40:58Z
Article
1940-087X
10.3791/57401
http://hdl.handle.net/10033/621571
https://www.jove.com/video/57401/rapid-vivo-assessment-adjuvant-s-cytotoxic-t-lymphocytes-generation
info:eu-repo/grantAgreement/EC/H2020/ 730964
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
embargoedAccess
Attribution-NonCommercial-ShareAlike 3.0 United States
136
Journal of Visualized Experiments
oai:repository.helmholtz-hzi.de:10033/6215882019-08-30T11:28:46Zcom_10033_620652col_10033_620666
2018-11-28T09:53:06Z
urn:hdl:10033/621588
Influenza-Activated ILC1s Contribute to Antiviral Immunity Partially Influenced by Differential GITR Expression.
Vashist, Neha
Trittel, Stephanie
Ebensen, Thomas
Chambers, Benedict J
Guzmán, Carlos A
Riese, Peggy
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
cross-talk
glucocorticoid-induced TNFR-related protein
influenza
innate lymphoid cell 1
regulation
Innate lymphoid cells (ILCs) represent diversified subsets of effector cells as well as immune regulators of mucosal immunity and are classified into group 1 ILCs, group 2 ILCs, and group 3 ILCs. Group 1 ILCs encompass natural killer (NK) cells and non-NK ILCs (ILC1s) and mediate their functionality via the rapid production of IFN-γ and TNF-α. The current knowledge of ILC1s mainly associates them to inflammatory processes. Much less is known about their regulation during infection and their capacity to interact with cells of the adaptive immune system. The present study dissected the role of ILC1s during early influenza A virus infection, thereby revealing their impact on the antiviral response. Exploiting in vitro and in vivo H1N1 infection systems, a cross-talk of ILC1s with cells of the innate and the adaptive immunity was demonstrated, which contributes to anti-influenza immunity. A novel association of ILC1 functionality and the expression of the glucocorticoid-induced TNFR-related protein (GITR) was observed, which hints toward a so far undescribed role of GITR in regulating ILC1 responsiveness. Overexpression of GITR inhibits IFN-γ production by ILC1s, whereas partial reduction of GITR expression can reverse this effect, thereby regulating ILC1 functionality. These new insights into ILC1 biology define potential intervention targets to modulate the functional properties of ILC1s, thus contributing toward the development of new immune interventions against influenza.
2018-11-28T09:53:06Z
2018-11-28T09:53:06Z
2018-01-01
Article
1664-3224
29623077
10.3389/fimmu.2018.00505
http://hdl.handle.net/10033/621588
http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Attribution-NonCommercial-ShareAlike 3.0 United States
Frontiers
Frontiers in immunology
oai:repository.helmholtz-hzi.de:10033/6216482019-08-30T11:33:54Zcom_10033_620659com_10033_620652col_10033_620666col_10033_620660
2019-01-15T13:45:26Z
urn:hdl:10033/621648
Signatures of T and B Cell Development, Functional Responses and PD-1 Upregulation After HCMV Latent Infections and Reactivations in Nod.Rag.Gamma Mice Humanized With Cord Blood CD34 Cells.
Theobald, Sebastian J
Khailaie, Sahamoddin
Meyer-Hermann, Michael
Volk, Valery
Olbrich, Henning
Danisch, Simon
Gerasch, Laura
Schneider, Andreas
Sinzger, Christian
Schaudien, Dirk
Lienenklaus, Stefan
Riese, Peggy
Guzman, Carlos A
Figueiredo, Constanca
von Kaisenberg, Constantin
Spineli, Loukia M
Glaesener, Stephanie
Meyer-Bahlburg, Almut
Ganser, Arnold
Schmitt, Michael
Mach, Michael
Messerle, Martin
Stripecke, Renata
BRICS, Braunschweiger Zentrum für Systembiologie, Rebenring 56,38106 Braunschweig, Germany.; HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
B cell class switch
HCMV
T cell maturation
humanized mice
linear discriminant analyses
optical imaging analyses
principal component analyses
reactivation
uman cytomegalovirus (HCMV) latency is typically harmless but reactivation can be largely detrimental to immune compromised hosts. We modeled latency and reactivation using a traceable HCMV laboratory strain expressing the Gaussia luciferase reporter gene (HCMV/GLuc) in order to interrogate the viral modulatory effects on the human adaptive immunity. Humanized mice with long-term (more than 17 weeks) steady human T and B cell immune reconstitutions were infected with HCMV/GLuc and 7 weeks later were further treated with granulocyte-colony stimulating factor (G-CSF) to induce viral reactivations. Whole body bio-luminescence imaging analyses clearly differentiated mice with latent viral infections vs. reactivations. Foci of vigorous viral reactivations were detectable in liver, lymph nodes and salivary glands. The number of viral genome copies in various tissues increased upon reactivations and were detectable in sorted human CD14+, CD169+, and CD34+ cells. Compared with non-infected controls, mice after infections and reactivations showed higher thymopoiesis, systemic expansion of Th, CTL, Treg, and Tfh cells and functional antiviral T cell responses. Latent infections promoted vast development of memory CD4+ T cells while reactivations triggered a shift toward effector T cells expressing PD-1. Further, reactivations prompted a marked development of B cells, maturation of IgG+ plasma cells, and HCMV-specific antibody responses. Multivariate statistical methods were employed using T and B cell immune phenotypic profiles obtained with cells from several tissues of individual mice. The data was used to identify combinations of markers that could predict an HCMV infection vs. reactivation status. In spleen, but not in lymph nodes, higher frequencies of effector CD4+ T cells expressing PD-1 were among the factors most suited to distinguish HCMV reactivations from infections. These results suggest a shift from a T cell dominated immune response during latent infections toward an exhausted T cell phenotype and active humoral immune response upon reactivations. In sum, this novel in vivo humanized model combined with advanced analyses highlights a dynamic system clearly specifying the immunological spatial signatures of HCMV latency and reactivations. These signatures can be merged as predictive biomarker clusters that can be applied in the clinical translation of new therapies for the control of HCMV reactivation.
2019-01-15T13:45:26Z
2019-01-15T13:45:26Z
2018-01-01
Article
- Front Immunol. 2018 Nov 22;9:2734. doi: 10.3389/fimmu.2018.02734. eCollection 2018.
1664-3224
30524448
10.3389/fimmu.2018.02734
http://hdl.handle.net/10033/621648
http://creativecommons.org/licenses/by-nc-sa/4.0/
Attribution-NonCommercial-ShareAlike 4.0 International
Frontiers
Frontiers in immunology
didl///col_10033_620666/100