A monoclonal antibody to a determinant of the rat T cell antigen receptor expressed by a minor subset of T cells

Abstract

A hybridoma producing the monoclonal antibody HIS42 was isolated from a fusion between spleen cells from a BALB/c mouse immunized with rat thymocytes and the fusion partner SP2/0. This antibody recognizes a minor subset of T cells in every haplotype of rat tested so far. The subpopulation of HIS42 positive T cells contains both CD4+ and CD8+ cells, in the same ratio as found in the peripheral T cell population. When bound to Sepharose beads, HIS42 induces T cell proliferation in the presence of interleukin-2. In contrast to lymph node T cells, a number of thymocytes were found to express HIS42 only in the cytoplasm or together with membrane expression. Most bright HIS42 surface labelled thymocytes were also positive for MRC OX-44, a marker predominantly identifying mature thymocytes. SDS-PAGE analyses of the membrane molecules immunoprecipitated by HIS42 show two bands on unreduced gels. One of these bands (85 kd) runs as two separate bands at 35 and 48 kd on reduction. The other much weaker broad band (approximately 100 kd) is hardly affected by reduced conditions. Taken together these data suggest that HIS42 is directed against a determinant on the rat T cell receptor for antigen, which is common to a small number of T cells.

Prevention of Type 1 Diabetes in the Rat With an Allele-Specific Anti–T-Cell Receptor Antibody

“…The hybridoma producing the His42 mouse anti-rat Vβ16 (IgG2b) mAb (19) was a gift from Dr. Thomas Hünig. Both antibodies were prepared as ascites and purified by affinity chromatography.…”

Identification of rat Tcrb-V8.2, 8.5, and 10 gene products by monoclonal antibodies

“…Currently, only one rat T-cell receptor (Tcr)-specific monoclonal antibody (mAb) with restricted expression among c~/3 T cells has been described (Kampinga et al 1989). Repertoire analysis in this species has therefore been limited to cDNA cloning and sequencing (Williams and Gutman 1989), polymerase chain reaction (PCR)-amplification (Gold and Bellgrau 1991), and RNAseprotection assays (Smith et al 1991).…”

“…Importantly, this assignment was in full agreement with PCR-typing of the RNA extracted from the T-cell hybrids reactive with the respective antibodies (Table 1). In this analysis, the above-mentioned Tcr-specific mAb, HIS42 (Kampinga et al 1989), reactive with about 10% of peripheral rat Tcells was included. Examination of HIS42-induced Tblasts indicated the presence of mRNA for Tcrb-V16 only (data not shown), but upon repetition an unambiguous assignment was not possible.…”

Analysis of TCR β chains in Lewis rats with experimental allergic encephalomyelitis. II. Vβ8.2+ T cells with limited CDR3 N region additions derive from the adult thymus

“…The following PEor FITC-conjugated mAb (obtained from PharMingen, San Diego, CA) were added to tubes in 1-? l volumes for 30 min, after which the cells were washed, resuspended in sorter buffer and run on a Coulter EPICS flow cytometer: mAb R73 to rat TCR g [20], R78 to V g 8.2 [21], HIS42 to V g 16 [22], OX38 to CD4 [23] and OX8 to CD8 [24].…”