Heather Medbury wrote: > >> I wanted to look at the human immune response to particular antigens. Iwanted to add > antgien to human leukocytes in culture and detect the immune response by flow and > proliferation assays. > I realise however that the immune response will involve a very small number of T cells > specific for the antigens, so I probably wouldnt be able to detect the response. 2 flow based assays for Ag specific cells are the intracellular cytokine assay and the tetramer method. The ICC assay involves adding the antigen to culture in the presence of a n inhibitor of secretion brefeldin A then permeabilizing the cellsa and labeling with antibodies for particular cytokines. Tetrameric MHC peptide complexes are loaded with the Ag of interest and specifically bind to cells which recognize it. The following references are some applications of these techniques. for the ICC assay: Nature Medicine, 5:518, 1999. J Infect Dis, 179:1124, 1999. for the tetramer assay: Science, 274:94, 1996. Science, 279:2103, 1998. J Virol, 74:1018,2000. Good luck, Tom ***************************************************************************** Thomas W. Mc Closkey, Ph. D. Director of Flow Cytometry, North Shore University Hospital Assistant Professor of Pediatrics, New York University School of Medicine Boas Marks Biomedical Research Center, 350 Community Drive Manhasset, Long Island, New York 11030 ph: 516-562-4844 [office], 516-562-1135/4641 [lab] fax: 516-562-2866 *****************************************************************************
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