Expansion of human lymphocyte populations expressing specific immune reactivities. II. A comparison of immune reactivities in human T lymphocyte lines derived from allogeneically primed cultures and maintained with lectins or conditioned medium.
In a previous paper, lectins were shown to allow a strong expansion of in vitro primed cells; among the lectins tested, PKW but not PHA (nor Con A) was found to reactivate CTLs. We then asked two further questions: Could one maintain a long term expansion of human T cells using iterative lectin stimulation? And, if so, could the immune reactivities, once expressed in the original primed cells, also be maintained? We report here that lectins remain potent mitogens for primed cells, and allow good expansion over many cycles, provided fresh irradiated cells (autologous or not to the responder) are added to the cultures. When PKW is used as the iterative mitogen, both the specific proliferation (PLT) and the specific cytotoxicity are maintained after each cycle. When PHA is the iterative stimulant, only the specific proliferation (PLT) is maintained, while no measurable cytolysis is present. However, if iteratively PHA stimulated primed cells are now cultured in the absence of PHA, even without additional specific stimulation, cytolytic activity is recovered from the cultures. In parallel, primed cells iteratively expanded using conditioned medium appear to retain both their specific lytic function and their specific proliferative function. Preliminary data suggest that the cloning efficiency when using lectins is higher than when using conditioned medium.
D002478:Cells, Cultured; D003470:Culture Media; D003602:Cytotoxicity, Immunologic; D000939:Epitopes; D006650:Histocompatibility Testing; D006801:Humans; D037102:Lectins / Q000494:pharmacology*; D008214:Lymphocytes / Q000145:classification / Q000276:immunology*; D010835:Phytohemagglutinins / Q000494:pharmacology; D011043:Pokeweed Mitogens / Q000494:pharmacology; D013601:T-Lymphocytes / Q000276:immunology*; D013997:Time Factors
D003470:Culture Media; D000939:Epitopes; D037102:Lectins; D010835:Phytohemagglutinins; D011043:Pokeweed Mitogens