Ultrasensitive quantification of TAP-dependent antigen compartmentalization in scarce primary immune cell subsets.
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
MetadataShow full item record
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.
CitationUltrasensitive quantification of TAP-dependent antigen compartmentalization in scarce primary immune cell subsets. 2015, 6:6199 Nat Commun
AffiliationTWINCORE, 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.
The following license files are associated with this item:
- Targeting tumour cells with defects in the MHC Class I antigen processing pathway with CD8+ T cells specific for hydrophobic TAP- and Tapasin-independent peptides: the requirement for directed access into the ER.
- Authors: Aladin F, Lautscham G, Humphries E, Coulson J, Blake N
- Issue date: 2007 Aug
- The transporter associated with antigen processing (TAP) is active in a post-ER compartment.
- Authors: Ghanem E, Fritzsche S, Al-Balushi M, Hashem J, Ghuneim L, Thomer L, Kalbacher H, van Endert P, Wiertz E, Tampé R, Springer S
- Issue date: 2010 Dec 15
- Exogenous peptides enter the endoplasmic reticulum of TAP-deficient cells and induce the maturation of nascent MHC class I molecules.
- Authors: Levitt JM, Howell DD, Rodgers JR, Rich RR
- Issue date: 2001 Apr
- Disruption of the association of 73 kDa heat shock cognate protein with transporters associated with antigen processing (TAP) decreases TAP-dependent translocation of antigenic peptides into the endoplasmic reticulum.
- Authors: Kamiguchi K, Torigoe T, Fujiwara O, Ohshima S, Hirohashi Y, Sahara H, Hirai I, Kohgo Y, Sato N
- Issue date: 2008 Feb
- Tapasin is required for efficient peptide binding to transporter associated with antigen processing.
- Authors: Li S, Paulsson KM, Chen S, Sjögren HO, Wang P
- Issue date: 2000 Jan 21