A method for producing monoclonal antibodies to human T-cell-receptor β-chain variable regions

Abstract

Study of the T-cell repertoire in humans has been hampered by the lack of monoclonal antibodies (mAbs) to the T-cell receptor (TCR) variable region (V) gene products. We describe a method for producing mAbs to the human TCR β-chain V (Vβ) gene products in which mice were immunized with a rat basophil cell line (RBL-2H3) transfected with the extracellular domain of the TCR heterodimer fused to the ζ chain of CD3. These cells acted as excellent immunogens for raising anti-TCR mAb and also formed the basis of a rapid screening assay. We generated mAbs against Vβ protein of the TCR, showed that these mAbs stained ≈1% of peripheral blood T cells, and further showed that the mAbs could stimulate proliferation of these T cells. We then characterized the mAbs by amplifying TCR cDNA derived from mAb-stimulated cells and sequencing the β chain. All clones sequenced used the Vβ7.1 chain, proving conclusively that the mAbs generated were specific for Vβ7.1 subfamily. This method generates mAbs to human TCR Vβ proteins efficiently and might allow production of a complete panel of mAbs directed against human TCR Bβ proteins.