Apoptotic elimination of V beta 8.2+ cells from the central nervous system during recovery from experimental autoimmune encephalomyelitis induced by the passive transfer of V beta 8.2+ encephalitogenic T cells.

A CD4+V beta 8.2+ T cell clone specific for the peptide 72-89 of guinea pig myelin basic protein (GMBP) was used to induce acute experimental autoimmune encephalomyelitis (EAE) in Lewis rats. To assess apoptosis in inflammatory cells infiltrating the central nervous system (CNS), we extracted cells from the spinal cord, enriched them for T cells and performed flow-cytometric analysis of their DNA stained with propidium iodide. The presence of apoptosis was confirmed by the demonstration of DNA fragmentation on gel electrophoresis. A gradual increase in the proportion of apoptotic cells was observed between 4 and 7 days after the transfer of the encephalitogenic T cells. The highest frequency of apoptotic cells (9.2 +/- 1.2%) was observed 7 days after cell transfer, when clinical recovery commenced. Passive transfer of ovalbumin-specific cells resulted in only a background level (0.8%) of apoptosis in the CNS. We conclude that the apoptotic process selectively eliminates autoreactive T cells from the CNS as: (a) there was a selective disappearance of disease-relevant CD5+V beta 8.2+ cells from the CNS during the course of EAE; (b) there was a decrease in the frequency of CNS-infiltrating T cells reactive to the GMBP 72-89 peptide during the course of EAE, and in a standard proliferation assay there was a loss of in vitro reactivity of CNS-infiltrating cells to this peptide, but not to a non-CNS antigen (ovalbumin); (c) simultaneous surface labeling and DNA analysis of CNS-infiltrating cells revealed that the frequency of V beta 8.2+ cells was about sevenfold higher in the apoptotic T cell population than in the normal (non-apoptotic) T cell population; and (d) we were unable to detect recirculation of the V beta 8.2+ cells to lymphoid organs after their frequency decreased in the CNS. The selective apoptotic elimination of autoreactive T cells from the target organ of this spontaneously resolving autoimmune disease may have implications for the understanding of the mechanism by which an autoimmune attack is terminated and for the design of therapeutic strategies to facilitate this process.