The use of primed cytotoxic T lymphocytes as a highly sensitive and accurate method for the detection of MHC disparate allogeneic cells.
The present studies have investigated the use of primed cytotoxic T cells (CTL) for the purpose of detecting the presence of an MHC disparate allogeneic cell population. The findings indicate that restimulation of primed CTL in the presence of activated T cell supernatants is an extremely sensitive method capable of detecting one allogeneic cell within a population containing 10,000 total cells (0.01%). In addition, this primed lymphocyte cytotoxicity assay (PLCA) was shown to be at least as accurate as flow cytometric analysis in determination of low numbers and percentages of allogeneic cells within the cell population being examined. Furthermore, the nature of the allogeneic mononuclear population did not influence the sensitivity or accuracy of this method. To examine this method's applicability for the detection of chimeric populations in vivo, spleen and thymic tissue from neonatally tolerized animals were examined by PLCA analysis. Allogeneic cells were readily detected in 100% of individual host spleen and thymuses tested. In total, the results showed that the PLCA is a highly reproducible and accurate method for the detection of extremely low numbers of allogeneic cells. This approach should be useful to monitor the presence of foreign cells in experimental and clinical situations in which individuals are exposed to allogeneic cell populations.
Richardson
J C
JC
Department of Microbiology and Immunology, University of Miami School of Medicine, FL 33101.
Levy
R B
RB
D000818:Animals; D000831:Animals, Newborn; D003601:Cytotoxicity Tests, Immunologic; D015395:Histocompatibility Antigens Class I / Q000032:analysis; D006801:Humans; D007108:Immune Tolerance; D008285:Major Histocompatibility Complex; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D015395:Histocompatibility Antigens Class I