FINE SPECIFICITY OF MONOCLONAL-ANTIBODIES DIRECTED AT HUMAN T-CELL RECEPTOR VARIABLE REGIONS - COMPARISON WITH OLIGONUCLEOTIDE-DRIVEN AMPLIFICATION FOR EVALUATION OF V-BETA EXPRESSION

Seven distinct anti-human T cell receptor (TcR) V region monoclonal antibodies (mAb) were generated by immunizing mice with either human T cell lines or transfected murine cells expressing human TcR Vbeta genes. The specificity of these reagents was determined as follows: T cells recognized by each mAb were purified from the peripheral blood of healthy donors and TcR transcripts expressed in these cells were analyzed using oligonucleotide-driven amplification and cDNA sequencing. Four mAb were found to delineate the Vbeta3, Vbeta8, Vbeta17 and Vbeta19 subfamilies, respectively. The remaining reagents recognize subsets within the Vbeta2, Vbeta5 and Vbeta13 subfamilies. Reactivity of the mAb with circulating T cells from 18 unrelated healthy individuals was determined. Limited variability was found from an individual to another. In four donors, mAb staining was compared to oligonucleotide-driven amplification for evaluation of Vbeta3, Vbeta8, Vbeta17 and Vbeta19 subfamily expression in the peripheral blood. Although the V gene subfamily-specific oligonucleotides used in this study belong to a carefully controlled series, our results show that this method does not give an accurate estimate of the percentage of peripheral T cells expressing a given TcR beta chain. The present data confirm the necessity to establish a complete set of well-characterized monoclonal reagents to study human T cell responses.