Production and characterization of monoclonal antibodies specific for the murine T cell receptor zeta chain.
The T cell receptor (TCR) comprises an antigen-specific alpha beta heterodimer non-covalently associated with the CD3 gamma delta epsilon and TCR zeta subunits. Both the CD3 and TCR zeta subunits are proposed to be responsible for the intracellular signal-transduction events. We report here the production of eight monoclonal antibodies (mAbs) that bind in an ELISA assay to a 113 amino acid synthetic peptide corresponding to the cytoplasmic domain of TCR zeta. Western blot analysis of anti-CD8 precipitates of lysates of transfectants expressing chimeric CD8/zeta constructs encoding increasing COOH-terminal truncations of TCR zeta indicates that four of these mAbs recognized the region of TCR zeta chain comprising the last 29 COOH-terminal residues. Thus, this region of TCR theta may encode an immunodominant epitope. Furthermore, one of these mAbs, G3, is capable of precipitating both non-phosphorylated and tyrosine phosphorylated TCR zeta. The G3 mAb should be useful for elucidating the structural and signalling characteristics of the TCR zeta chain.
van Oers
N S
NS
Howard Hughes Medical Institute, San Francisco, CA 94143-0724.
Teh
S J
SJ
Irving
B A
BA
Tiong
J
J
Weiss
A
A
Teh
H S
HS
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D015153:Blotting, Western; D016827:CD8 Antigens / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology; D015458:Leukemia, T-Cell; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D016827:CD8 Antigens; D000939:Epitopes; D007074:Immunoglobulin G; D008565:Membrane Proteins; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; C054979:antigen T cell receptor, zeta chain