Human cytotoxic T lymphocytes. I. Limiting-dilution analysis of alloreactive cytotoxic T-lymphocyte precursor frequencies.
A limiting-dilution system was established to measure the frequency of alloreactive cytotoxic T-lymphocyte precursors (CTL-p) in human peripheral blood T cells. Culture medium supplemented with recombinant interleukin-2 enabled clonal expansion of all CTL-p stimulated by allogeneic peripheral blood or spleen cells. The range of CTL-p frequencies in fully HLA-mismatched responder-stimulator combinations was 1:240 to 1:1230. Split-well analysis of individual microwells showed that the cytotoxic T-cell clones generated under limiting-dilution conditions showed exquisite specificity for the stimulating alloantigens. Alloreactive CTL-p were enriched in the OKT4- T-cell subset. This limiting-dilution system was highly reproducible and can thus be applied to investigate human cytotoxic T-cell precursor frequencies in various clinically relevant situations.
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Kabelitz
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Herzog
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Zanker
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Wagner
D002478:Cells, Cultured; D000939:Epitopes; D006680:HLA Antigens / Q000032:analysis; D006801:Humans; D007519:Isoantigens / Q000276:immunology; D007700:Kinetics; D007958:Leukocyte Count / Q000379:methods; D013601:T-Lymphocytes / Q000145:classification; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology*
D000939:Epitopes; D006680:HLA Antigens; D007519:Isoantigens