Specific and non-specific components in the triggering of proliferative and cytotoxic responses of T lymphocytes with different cell density |
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Authors: | M T Bejarano M G Masucci E Klein |
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Institution: | Department of Tumor Biology, Karolinska Institutet, Stockholm, Sweden |
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Abstract: | We hve analyzed the functional behavior of lymphocyte subsets separated on the basis of cell density. Low and high density subpopulations were cultured in FCS, alone or with allogeneic irradiated PBL, and then examined for proliferation and cytotoxic activity against autologous (responder) and allogeneic (stimulator) PHA-induced blasts, K562 and Daudi. In the high density subset proliferation and generation of anti-K562 and anti-Daudi effects were induced by FCS and to higher extent by allospecific stimulation. Exposure to alloantigens induced allospecific cytotoxicity. Autologous PHA blasts were not affected. The results with the low density subset differed. Independently of the type of stimulus imposed, the low density fraction showed little if any proliferation, but its cytotoxic activity was stronger against all targets tested. In some of the experiments, anti-alloblast cytotoxicity was generated in the control cultures. Thus, polyclonal activation induced by FCS triggered in this fraction allospecific cytotoxicity. In this subset, the effect against allogeneic PHA blasts comprised a specific and a non-specific component because autologous PHA blasts were also lysed. Limiting dilution analysis involving allostimulation showed higher frequency of cytotoxic precursors in the low density subset. Split minicultures were tested for lysis of auto- and allogeneic blasts. Alloreactive cultures that did not lyse the autologous target were more frequent in the cultures initiated with the high density cells. There was no conclusive evidence for the existence of autoreactive cultures that did not lyse the allogeneic blasts. |
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Keywords: | CTL cytotoxic T lymphocytes CTL-P CTL precursors FCS fetal calf serum HS human serum PHA phytohemagglutinin LGL large granular lymphocyte CC-s control cultures MLC-s mixed lymphocyte cultures SRBC sheep erythrocytes PBL peripheral blood lymphocytes LDA limiting dilution analysis |
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