Ultrasensitive quantification of TAP-dependent antigen compartmentalization in scarce primary immune cell subsets.

2.50
Hdl Handle:
http://hdl.handle.net/10033/346516
Title:
Ultrasensitive quantification of TAP-dependent antigen compartmentalization in scarce primary immune cell subsets.
Authors:
Fischbach, Hanna; Döring, Marius; Nikles, Daphne; Lehnert, Elisa; Baldauf, Christoph; Kalinke, Ulrich ( 0000-0003-0503-9564 ) ; Tampé, Robert
Abstract:
Presentation of peptides on major histocompatibility complex class I (MHC I) is essential for the establishment and maintenance of self-tolerance, priming of antigen-specific CD8(+) T cells and the exertion of several T-cell effector functions. Cytosolic proteasomes continuously degrade proteins into peptides, which are actively transported across the endoplasmic reticulum (ER) membrane by the transporter associated with antigen processing (TAP). In the ER lumen antigenic peptides are loaded onto MHC I, which is displayed on the cell surface. Here we describe an innovative flow cytometric approach to monitor time-resolved ER compartmentalization of antigenic peptides. This assay allows the analysis of distinct primary human immune cell subsets at reporter peptide concentrations of 1 nM. Thus, this ultrasensitive method for the first time permits quantification of TAP activity under close to physiological conditions in scarce primary cell subsets such as antigen cross-presenting dendritic cells.
Affiliation:
TWINCORE, Centre for Experimental and Clinical Infection Research, a joint venture between the Helmholtz-Centre for Infection Research and the Hannover Medical School, Feodor-Lynen Str. 7-9, 30625 Hannover, Germany.
Citation:
Ultrasensitive quantification of TAP-dependent antigen compartmentalization in scarce primary immune cell subsets. 2015, 6:6199 Nat Commun
Journal:
Nature communications
Issue Date:
2015
URI:
http://hdl.handle.net/10033/346516
DOI:
10.1038/ncomms7199
PubMed ID:
25656091
Type:
Article
Language:
en
ISSN:
2041-1723
Appears in Collections:
publications of the department of experimental infection research ([TC] EXPI)

Full metadata record

DC FieldValue Language
dc.contributor.authorFischbach, Hannaen
dc.contributor.authorDöring, Mariusen
dc.contributor.authorNikles, Daphneen
dc.contributor.authorLehnert, Elisaen
dc.contributor.authorBaldauf, Christophen
dc.contributor.authorKalinke, Ulrichen
dc.contributor.authorTampé, Roberten
dc.date.accessioned2015-03-11T09:41:46Zen
dc.date.available2015-03-11T09:41:46Zen
dc.date.issued2015en
dc.identifier.citationUltrasensitive quantification of TAP-dependent antigen compartmentalization in scarce primary immune cell subsets. 2015, 6:6199 Nat Communen
dc.identifier.issn2041-1723en
dc.identifier.pmid25656091en
dc.identifier.doi10.1038/ncomms7199en
dc.identifier.urihttp://hdl.handle.net/10033/346516en
dc.description.abstractPresentation of peptides on major histocompatibility complex class I (MHC I) is essential for the establishment and maintenance of self-tolerance, priming of antigen-specific CD8(+) T cells and the exertion of several T-cell effector functions. Cytosolic proteasomes continuously degrade proteins into peptides, which are actively transported across the endoplasmic reticulum (ER) membrane by the transporter associated with antigen processing (TAP). In the ER lumen antigenic peptides are loaded onto MHC I, which is displayed on the cell surface. Here we describe an innovative flow cytometric approach to monitor time-resolved ER compartmentalization of antigenic peptides. This assay allows the analysis of distinct primary human immune cell subsets at reporter peptide concentrations of 1 nM. Thus, this ultrasensitive method for the first time permits quantification of TAP activity under close to physiological conditions in scarce primary cell subsets such as antigen cross-presenting dendritic cells.en
dc.language.isoenen
dc.titleUltrasensitive quantification of TAP-dependent antigen compartmentalization in scarce primary immune cell subsets.en
dc.typeArticleen
dc.contributor.departmentTWINCORE, Centre for Experimental and Clinical Infection Research, a joint venture between the Helmholtz-Centre for Infection Research and the Hannover Medical School, Feodor-Lynen Str. 7-9, 30625 Hannover, Germany.en
dc.identifier.journalNature communicationsen

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