Thymic nurse cell rescue of early CD4+CD8+ thymocytes from apoptosis

We have now developed temperature sensitive lines of thymic nurse cells (TNCs), using the SV40 viral mutant tsA58, that maintain the ability to selectively internalize a subpopulation of alpha beta TCR+CD4+CD8+ thymocytes in vitro. One line, tsTNC-1, was shown to be able to rescue a subset of CD4+CD8+ thymocytes from programmed cell death at 32 degrees C, the temperature at which binding and internalization were detected. Rescue was significantly diminished at 38 degrees C, the temperature at which thymocyte binding was not observed. The rescued population of thymocytes showed a reduced level of apoptosis as measured by the DNA fragmentation assay. TNC rescue resulted in a shift of CD4+CD8+ thymocytes from immature TCRlow PNArhigh cells to the more mature TCRint PNArlow phenotype but no changes in cell surface levels of HSA nor CD69 were detected. The rescue activity of tsTNC-1 cells at 32 degrees C was significantly reduced with the addition of antibodies to either class I or class II MHC antigens. These results suggest that we have established TNC lines, using the SV40 viral mutant tsA58, that have the ability to rescue a subset of the TNC interactive thymocyte population from programmed cell death. The thymocyte population rescued by TNCs matures to a phenotype within the double positive stage of development that is indicative of positive selection.