Phosphorylation of the CD8 antigen on cytotoxic human T cells in response to phorbol myristate acetate or antigen-presenting B cells.
We have used the monoclonal antibody OKT8 to demonstrate that the cluster designation (CD)8 antigen on cytotoxic human T cells undergoes a previously unreported protein modification. Immunoprecipitation of CD8 with OKT8 from a CD8+ and CD4+ T cell line prelabeled with [32P]phosphate demonstrates that CD8 can be constitutively labeled with phosphate. CD8 undergoes rapid and intense phosphorylation in serine upon addition of phorbol myristate acetate to the cells. CD8 phosphorylation is induced upon addition of heterologous, Epstein-Barr virus-transformed B cells, which cause proliferation and are target cells for a cytotoxic CD8+CD4+ T cell line and a CD8+CD4- T cell clone. Phosphorylation induced by targets is dose-dependent, rapid, and followed by a fast dephosphorylation. Epstein-Barr virus-transformed B cells that do not induce proliferation of and are not targets for the CD8+CD4+ line and the CD8+CD4- clone do not induce CD8 phosphorylation. Cloned CD8+CD4- cells that proliferate in response to target cells, but lyse them only in the presence of lectin do not undergo target cell-induced CD8 phosphorylation. These data suggest that induction of CD8 phosphorylation is antigen-specific and is coincident with the cytotoxic response. Finally, preincubation of effector and target cells with an antibody to a monomorphic determinant of major histocompatibility complex class I antigens reduces target-induced CD8 phosphorylation to a greater extent than antibody to a major histocompatibility complex class II subregion (DR) monomorphic determinant, reinforcing the notion that major histocompatibility complex class I antigens interact with CD8+ cells.
D000911:Antibodies, Monoclonal / Q000276:immunology; D000938:Antigen-Presenting Cells / Q000276:immunology*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis*; D001402:B-Lymphocytes / Q000276:immunology*; D000001:Calcimycin / Q000494:pharmacology; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D006801:Humans; D010750:Phosphoproteins / Q000096:biosynthesis / Q000276:immunology; D010766:Phosphorylation; D010835:Phytohemagglutinins / Q000494:pharmacology; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D000911:Antibodies, Monoclonal; D000945:Antigens, Differentiation, T-Lymphocyte; D010750:Phosphoproteins; D010835:Phytohemagglutinins; D000001:Calcimycin; D013755:Tetradecanoylphorbol Acetate