Fine specificity of monoclonal antibodies directed at human T cell receptor variable regions: Comparison with oligonucleotide‐driven amplification for evaluation of Vβ expression

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Abstract
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 Vβ 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 Vβ3, Vβ8,Vβ17 and Vβ19 subfamilies, respectively. The remaining reagents recognize subsets within the Vβ2,Vβ5 and Vβ13 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 Vβ3, Vβ8,Vβ17 and Vβ19 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 β chain. The present data confirm the necessity to establish a complete set of well‐characterized monoclonal reagents to study human T cell responses.