Human T lymphocyte clones: influence of culture conditions and optimization of proliferative assays.
Many CD4+ human T lymphocyte clones (TLC) are found not to proliferate against appropriate stimulating cells, and many lose this capacity during culture. This may be due, not to a defect in the recognition of the antigen, but to an inability to produce sufficient amounts of interleukin 2 (IL-2) for autocrine growth, since specific HLA-restricted proliferative responses could be induced in 'non-proliferative' clones by the addition of exogenous IL-2 or phorbol myristate acetate (PMA). Of various factors tested during expansion procedures of the clones, the proliferative capacity could only be restored by changing the stimulatory cells from B lymphoblastoid cell lines (B-LCL) to peripheral blood mononuclear cells (PBM). The cytotoxicity of the TLC was found to be independent of its proliferative capacity. After restoration of the proliferative capacity, a mouse B lymphoma cell line transfected with the appropriate HLA DQA and DQB genes was still not able to induce proliferation in the absence of exogenous IL-2. We conclude that (1) 'non-proliferative' TLC may recognize their targets, but fail to proliferate due to temporary lack of IL-2 production, and (2) even 'proliferative' T cells may fail to respond to certain target cells carrying the specific antigen, such as a murine transfectant, in the absence of exogenous IL-2.
Lundin
K E
KE
Institute of Transplantation Immunology, National Hospital, Oslo, Norway.
Bosnes
V
V
Gaudernack
G
G
D000818:Animals; D000911:Antibodies, Monoclonal; D002999:Clone Cells; D003470:Culture Media; D003601:Cytotoxicity Tests, Immunologic; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D013601:T-Lymphocytes / Q000187:drug effects*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D003470:Culture Media; D007376:Interleukin-2; D013755:Tetradecanoylphorbol Acetate