Induction of CD3 delta epsilon omega by phorbol 12-myristate 13-acetate.
The effect of phorbol 12-myristate 13-acetate (PMA) on the synthesis, assembly and processing of the components of the T cell receptor (TcR) was studied with special focus on the CD3 omega chain. Treatment of the human leukemic T cell line Jurkat with PMA increased the synthesis of the Ti alpha, CD3 gamma and CG3 zeta chains two- to threefold and the synthesis of Ti beta and CD3 delta epsilon omega complexes five- to sevenfold as assessed by metabolic labeling, immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by scanning densitometry. The amount of total assembled TcR complexes increased approximately threefold and the maturation of the TcR was not affected as determined by analysis of oligosaccharide side chain processing in the Golgi apparatus. Activation of Jurkat cells with anti-CD3 monoclonal antibody, calcium ionophore, or mitogenic lectins did not affect the synthesis of the TcR components. In other cells studied (the human leukemic T cell line CEM, a panel of variants of the Jurkat T cell line and peripheral blood mononuclear cells) PMA also increased the synthesis of the TcR components. However, for all cell lines studied the amount of TcR complexes expressed on the cell surface was decreased after 16 h of PMA treatment. Based on these results we propose a role of CD3 omega in retention of TcR complexes. From PMA-treated CEM cells more than 50-fold the amount of CD3 delta epsilon omega complexes was immunoprecipitated as compared to the amount obtained from untreated Jurkat cells, and these observations indicate that the CEM cell line may be a qualified candidate for purification of CD3 omega.
Vangsted
A
A
Department of Tumor Cell Biology, Danish Cancer Society, Copenhagen.
Neisig
A
A
Wallin
H
H
Zeuthen
J
J
Geisler
C
C
D000643:Ammonium Chloride / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000276:immunology; D017252:CD3 Complex / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D015759:Ionomycin / Q000494:pharmacology; D046911:Macromolecular Substances; D016853:Muromonab-CD3 / Q000276:immunology; D010835:Phytohemagglutinins / Q000494:pharmacology; D011485:Protein Binding; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D046911:Macromolecular Substances; D016853:Muromonab-CD3; D010835:Phytohemagglutinins; D016693:Receptors, Antigen, T-Cell, alpha-beta; D000643:Ammonium Chloride; D015759:Ionomycin; D013755:Tetradecanoylphorbol Acetate