The Journal of Immunology, Vol 144, Issue 3 816-823, Copyright © 1990 by American Association of Immunologists

ARTICLES

Development of precytotoxic T cells in cyclosporine-suppressed mixed lymphocyte reactions

JW Hooton, CL Miller, CD Helgason, RC Bleackley, S Gillis and V Paetkau
Department of Biochemistry, University of Alberta, Edmonton, Canada.

Cyclosporine (CsA) blocked the generation of cytolytic activity in a primary MLR of mouse spleen cells. As expected from the known mechanism of action of this drug, it also blocked the accumulation of IL-2 during the MLR. Addition of human rIL-2 did not overcome the inhibition of CTL generation, even when it was added daily to keep its level similar to that produced in a normal MLR. Daily addition was necessary, because the CsA-inhibited MLR consumed IL-2, either by utilization or degradation. The outcome of a 5-day MLR in the presence of CsA (CsA- MLR) depended on whether or not IL-2 was continuously present. In the presence of IL-2, there was no generation of CTL activity, probably because such cultures contained IL-2-dependent suppressive elements described previously. However, when day 5 CsA-MLR cells generated in the absence of IL-2 were washed and recultured with human rIL-2, there was a burst of CTL activity, with a more than 50-fold increase in alloantigen-specific cytotoxicity within 24 to 48 h. This increase is not explainable simply by the proliferation of existing effector CTL. The noncytotoxic cells produced in an MLR in the presence of CsA, and which can be rapidly activated to cytotoxic effector cells by IL-2, are termed "precursor-effector CTL" (peCTL). They could be detected by day 3 of a primary CsA-MLR culture. Their conversion to effector CTL by IL- 2 was not inhibited by CsA. Exposure of peCTL to IL-4 also generated CTL activity, to a somewhat lesser degree than IL-2, but the IL-4- induced activation was inhibited by CsA, suggesting that it depended on the induction of another CsA-sensitive lymphokine. The intracellular levels of mRNA encoding the CTL-specific serine esterases CCP1 and CCP2 (granzymes B and C, respectively) increased rapidly during the IL-2- driven conversion of peCTL to effector CTL. This study demonstrates that in the presence of CsA precursors for CTL can accumulate, and that these can be rapidly converted to cytotoxic effector cells by IL-2.

This article has been cited by other articles:

				T. J. Dengler and J. S. Pober Human Vascular Endothelial Cells Stimulate Memory But Not Naive CD8+ T Cells to Differentiate into CTL Retaining an Early Activation Phenotype J. Immunol., May 15, 2000; 164(10): 5146 - 5155. [Abstract] [Full Text] [PDF]

				T. Guillaume, D. B. Rubinstein, and M. Symann Immune Reconstitution and Immunotherapy After Autologous Hematopoietic Stem Cell Transplantation Blood, September 1, 1998; 92(5): 1471 - 1490. [Full Text] [PDF]