title: Production and characterization of a novel monoclonal antibody against phosphorylated T cell receptor zeta chain.
abstract: One of the earliest events following T cell receptor (TCR) triggering is the activation of the protein kinase Lck and induction of tyrosine phosphorylation of zeta, the major signal transduction subunit of the T cell receptor complex. Here we report the generation and characterization of a monoclonal antibody specific for human phosphozeta. The antibody was produced by immunizing mice with a truncated recombinant form of human zeta together with the Lck enzyme. The C415.9A antibody recognizes recombinant as well as cellular phosphozeta but is unreactive with unphosphorylated zeta or other tyrosine phosphorylated proteins. Using this antibody, we have demonstrated aberrant TCR-zeta tyrosine phosphorylation in Jurkat T cell transduction mutants. Therefore, this antibody can be used to elucidate T cell signal transduction mechanisms by analyzing and monitoring tyrosine phosphorylation of zeta in vitro and in vivo directly. Furthermore, this antibody could find application in the analysis of abnormal T cell signaling in autoimmune disease, cancer, and immunodeficiency disorders.
authors.affiliation: Institute for Bone and Joint Disorders and Cancer, Bayer Research Center, West Haven, CT 06516, USA.
authors.forename: P J
authors.initials: PJ
authors.lastname: Conrad
authors.forename: T
authors.initials: T
authors.lastname: Seng
authors.forename: J
authors.initials: J
authors.lastname: Barbosa
authors.forename: H
authors.initials: H
authors.lastname: Vasavada
authors.forename: M L
authors.initials: ML
authors.lastname: Blue
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes; D018414:CD8-Positive T-Lymphocytes; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D009154:Mutation; D010750:Phosphoproteins / Q000276:immunology*; D010766:Phosphorylation; D011233:Precipitin Tests; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D015398:Signal Transduction / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; D010750:Phosphoproteins; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins; C054979:antigen T cell receptor, zeta chain