Preparation and characterization of a “pan-reactive” rabbit anti-mouse T-cell receptor antiserum

Abstract

A pan-reactive xenoantiserum to the mouse T-cell receptor was prepared by immunization of a rabbit with affinity purified mouse T-cell receptor material. The T-cell receptor of the chicken ovalbumin/IA d specific T-cell hybridoma, DO-11.10, was isolated by affinity chromatography using the clone-specific monoclonal antibody, KJ1-26. Immunoprecipitation with the rabbit antiserum and subsequent SDS-PAGE analysis of the material precipitated from lysates of surface radioiodinated T cells revealed the heterodimeric structure characteristic of the T-cell receptor from virtually every T-cell source examined. Flow cytofluorometric analysis of normal peripheral T cells and mature thymocytes of BALB/c and SJL mice indicated that most all T cells bear antigenic determinants recognized by the rabbit anti-mouse T-cell receptor antibodies. The AKR thymoma, BW5147, a common fusion parent used to generate functional T-cell hybridomas, notably lacks surface expression of a T-cell receptor molecule.