T cell derived proteins from normal human sera and their relationship to T cell antigen binding molecules.
We have used procedures which have been developed to isolate murine T cell antigen binding molecules (TABM) in order to isolate TABM from normal human sera. To begin purification, ammonium sulfate (NH4)2SO4 was added to human serum and precipitated protein was dissolved in low salt buffer and resolved by ion-exchange chromatography on carboxymethylcellulose (CM). The most strongly CM nonadherent fraction was absorbed with anti-human albumin and anti-human immunoglobulin (Ig) antibodies conjugated to Sepharose beads. The resulting nonadsorbed 110,000, 70,000 and 45,000 Mr polypeptides were reactive in ELISA with a rabbit antiserum produced against non-Ig, anti-specific molecules of rhesus monkeys. These proteins possess alpha mobility upon immunoelectrophoresis and represent 0.02 to 0.05% of total serum protein. In addition, these proteins are bound by an antiserum made against a synthetic peptide corresponding to the J region of the TcR beta chain. We have made R28, a rabbit antiserum against these serum proteins which binds specifically to tetanus-specific polypeptides obtained from the culture supernatant of human T cell lines specific for tetanus. This antiserum also binds to proteins isolated from T cell but not B cell lines, and T cell proteins are able to inhibit the binding of R28 to the human serum polypeptides. The results suggest that the proteins isolated from normal human sera are T cell antigen binding molecules.
DiBrino
M
M
Department of Pathology, University of Connecticut Health Center, Farmington 06030.
Ravindran
B
B
Cone
R E
RE
D000818:Animals; D000941:Antigens / Q000378:metabolism*; D001798:Blood Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D008970:Molecular Weight; D011817:Rabbits; D013601:T-Lymphocytes / Q000737:chemistry*; D013745:Tetanus Toxoid / Q000276:immunology
D000941:Antigens; D001798:Blood Proteins; D007106:Immune Sera; D013745:Tetanus Toxoid