Sensitive and viable identification of antigen-specific CD8+ T cells by a flow cytometric assay for degranulation. Export

@article{citeulike:851400, abstract = {Flow cytometric detection of antigen-specific CD8+ T cells has previously been limited to MHC-class I tetramer staining or intracellular cytokine production, neither of which measure the cytolytic potential of these cells. Here we present a novel technique to enumerate antigen-specific CD8+ T cells using a marker expressed on the cell surface following activation induced degranulation, a necessary precursor of cytolysis. This assay measures the exposure of CD107a and b, present in the membrane of cytotoxic granules, onto the cell surface as a result of degranulation. Acquisition of cell surface CD107a and b is associated with loss of intracellular perforin and is inhibited by colchicine, indicating that exposure of CD107a and b to the cell surface is dependent on degranulation. CD107a and b are expressed on the cell surface of CD8+ T cells following activation with cognate peptide, concordant with production of intracellular IFNgamma. Finally, CD107-expressing CD8+ T cells are shown to mediate cytolytic activity in an antigen-specific manner. Measurement of CD107a and b expression can also be combined with MHC-class I tetramer labeling and intracellular cytokine staining to provide a more complete assessment of the functionality of CD8+T cells expressing cognate T cell receptors (TCR).}, address = {Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 40 Convent Drive, Bethesda, MD 20892, USA. mbetts@mail.nih.gov}, author = {Betts, M. R. and Brenchley, J. M. and Price, D. A. and De Rosa, S. C. and Douek, D. C. and Roederer, M. and Koup, R. A.}, citeulike-article-id = {851400}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/14580882}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=14580882}, comment = {flipped through it}, day = {1}, issn = {0022-1759}, journal = {J Immunol Methods}, keywords = {assay, lamp}, month = {October}, number = {1-2}, pages = {65--78}, posted-at = {2007-08-14 02:20:54}, priority = {2}, title = {Sensitive and viable identification of antigen-specific CD8+ T cells by a flow cytometric assay for degranulation.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/14580882}, volume = {281}, year = {2003} }

TY - JOUR ID - citeulike:851400 L3 - citeulike-article-id:851400 N2 - Flow cytometric detection of antigen-specific CD8+ T cells has previously been limited to MHC-class I tetramer staining or intracellular cytokine production, neither of which measure the cytolytic potential of these cells. Here we present a novel technique to enumerate antigen-specific CD8+ T cells using a marker expressed on the cell surface following activation induced degranulation, a necessary precursor of cytolysis. This assay measures the exposure of CD107a and b, present in the membrane of cytotoxic granules, onto the cell surface as a result of degranulation. Acquisition of cell surface CD107a and b is associated with loss of intracellular perforin and is inhibited by colchicine, indicating that exposure of CD107a and b to the cell surface is dependent on degranulation. CD107a and b are expressed on the cell surface of CD8+ T cells following activation with cognate peptide, concordant with production of intracellular IFNgamma. Finally, CD107-expressing CD8+ T cells are shown to mediate cytolytic activity in an antigen-specific manner. Measurement of CD107a and b expression can also be combined with MHC-class I tetramer labeling and intracellular cytokine staining to provide a more complete assessment of the functionality of CD8+T cells expressing cognate T cell receptors (TCR). IS - 1-2 JF - J Immunol Methods SN - 0022-1759 AD - Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 40 Convent Drive, Bethesda, MD 20892, USA. mbetts@mail.nih.gov EP - 78 TI - Sensitive and viable identification of antigen-specific CD8+ T cells by a flow cytometric assay for degranulation. VL - 281 SP - 65 KW - assay KW - lamp N1 - flipped through it AU - Betts, MR AU - Brenchley, JM AU - Price, DA AU - De Rosa, SC AU - Douek, DC AU - Roederer, M AU - Koup, RA PY - 2003/10/01/ UR - http://view.ncbi.nlm.nih.gov/pubmed/14580882 ER -