Abstract
Postthymic T cell receptor (TCR)-mediated cell death offers the potential for creating antigen-specific transplant tolerance analogous to thymic clonal deletion. Murine specific CD4+ cell were rigorously purified by: (a) adherent cell depletion and (b) magnetic bead/monoclonal antibody (mAb) depletion of macrophages, Ia+ cells, mu-chain+ cells, NK cells, and CD8+ T cells. CD4+s were typically > 95% pure by flow cytometry. Resting CD4+s stimulated by plastic-immobilized anti-TCR/CD3 mAb wee shown to die in the absence of exogenous interleukin (IL)-2. Blasting CD4+s showed dose-dependent cell death upon religation of TCR/CD3 in the presence of IL-2; however, withdrawal of IL-2 from blasting CD4+s also resulted in cell death. Cell death was shown to be apoptotic by flow cytometry DNA content analysis. Anti-CD28 mAb, co-immobilized with anti-TCR/CD3 mAb, inhibited cell death of resting CD4+s in the absence of exogenous IL-2; however, anti-CD28 mAb showed minimal cell death inhibition of CD4+ blasts when TCR/CD3 was religated. In contrast, splenic adherent cells effectively inhibited cell death of blasting CD4+s induced by TCR/CD3 mAb religation. We conclude that TCR-mediated programmed cell death of highly purified splenic CD4+s is...Continue Reading
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