Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we'll assume that you are happy to receive all cookies and you won't see this message again. Click 'Find out more' for information on how to change your cookie settings.

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 beta-chain V (V beta) 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 lambda 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 V beta protein of the TCR, showed that these mAbs stained approximately 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 beta chain. All clones sequenced used the V beta 7.1 chain, proving conclusively that the mAbs generated were specific for V beta 7.1 subfamily. This method generates mAbs to human TCR V beta proteins efficiently and might allow production of a complete panel of mAbs directed against human TCR V beta proteins.

Type

Journal article

Journal

Proceedings of the National Academy of Sciences of the United States of America

Publication Date

11/1993

Volume

90

Pages

10454 - 10458

Addresses

Institute of Molecular Medicine, John Radcliffe Hospital, Oxford, United Kingdom.

Keywords

Hybridomas, Tumor Cells, Cultured, Animals, Humans, Rats, Leukemia, Basophilic, Acute, Receptors, Antigen, T-Cell, alpha-beta, Recombinant Proteins, Antibodies, Monoclonal, Precipitin Tests, Transfection, Lymphocyte Activation, Amino Acid Sequence, Molecular Sequence Data