Abstract
We have studied the diversity of the expressed human T-cell receptor (TCR) β-chain repertoire by analysis of mRNA from unstimulated peripheral blood T-cells. The anchored-polymerase chain reaction (PCR) was used to isolate TCRB transcripts. Of 20 full or near full-length functional transcripts sequenced, two were novel TCRVB gene segments. They have strong sequence similarities to the known TCRBV5, and 8 subfamilies. Southern blot analysis and sequence-specific oligonucleotide hybridization confirmed: a) that these sequences are present in genomic DNA; b) their relationship to the known TCRBV families. A TCRBV sequence similar to a recently identified novel TCRBV24 subfamily was also found. We show by southern blotting that this sequence forms a single member subfamily, and by deletion analysis of T-cell lines, we have mapped this sequence to lie between the genes which encode the TCRBV8.1 and TCRBV5.3 gene segments. The results show that the anchored PCR is a powerful tool in the analysis of the TCR repertoire, which may contain more V gene segments than previously defined.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Concannon, P., Pickering, L. A., Kung, P., and Hood, L.: Diversity and structure of human T-cell receptor beta-chain variable region genes. Proc Natl Acad Sci USA 83: 65986602, 1986
Davis, M. M. and Bjorkman, P. J.: T-cell antigen receptor genes and T-cell recognition. Nature 334: 395–402, 1988
Ferradini, L., Roman-Roman, S., Azocar, J., Michalaki, H., Triebel, F., and Hercend, T.: Studies on the human T-cell receptor alpha/beta variable region genes II. Identification of four additional Vβ subfamilies. Eur J Immunol 21: 935–942, 1991
Kabat, E. A., Wu, T. T., Reid-Miller, M., Perry, H. M., and Gottesman, K. S.: Sequences of Proteins of Immunological Interest, 4th edition, U.S. Department of Health and Human Services, National Institutes of Health, Bethesda, MD, 1987
Kimura, N., Toyonaga, B., Yoshikai, Y., Triebel, F., Debre, P., Minden, M. D., and Mak, T. W.: Sequences and diversity of human T-cell receptor β chain variable region genes. J Exp Med 164: 739–750, 1986
Lai, E., Concannon, P., and Hood, L.: Conserved organization of the human and murine T-cell receptor beta-gene families. Nature 331: 543–546, 1988
Leiden, J. M. and Strominger, J. L.: Generation of diversity of the beta chain of the human T-lymphocyte receptor for antigen. Proc Natl Acad Sci USA 83: 4456–4460, 1986
Loh, E. Y., Elliott, J. F., Cwirla, S., Lanier, L. L., and Davis, M. M.: Polymerase chain reaction with single sided specificity: Analysis of T-cell receptor σ chain. Science 243: 217–220, 1989
Robinson, M. A.: The human T-cell receptor β-chain gene complex contains at least 57 variable gene segments. J Immunol 146: 4392–4397, 1991
Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., and Erlich, H. A.: Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239: 487, 1988
So, A. K. L., Lindsay, J., Bodmer, J., and Trowsdale, J.: Molecular variation of human major histocompatibility complex DQw3β-chains. J Immunol 139: 3506–3511, 1987
Tillinghast, J. P., Behlke, M. A., and Loh, D. Y.: Structure and diversity of the human T-cell receptor beta chain variable region genes. Science 233: 879–883, 1986
Toyonaga, B., Yoshikai, Y., Vadasz, V., Chin, B., and Mak, T. W.: Organization and sequences of the diversity, joining, and constant region genes of the human T-cell receptor beta-chain. Proc Natl Acad Sci USA 82: 8624–8628, 1985
Toyonaga, B. and Mak, T. W.: Genes of the T-cell antigen receptor in normal and malignant T cells. Annu Rev Immunol 5: 585–620, 1987
Wilson, R. K., Lai, E., Concannon, P., Barth, R. K., and Hood, L. E.: Structure, organization and polymorphism of murine and human T-cell receptor alpha and beta chain gene families. Immunol Rev 101: 149–172, 1988
Wilson, R. K., Lai, E., Kim, L. D. H., and Hood, L.: Sequence and expression of a novel human T-cell receptor beta-chain variable gene segment subfamily. Immunogenetics, 32: 406–412, 1990
Author information
Authors and Affiliations
Additional information
The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession numbers M73463, M73464, and M73465.
Address correspondence and offprint request to: A. K. So.
Rights and permissions
About this article
Cite this article
Lunardi, C., Marguerie, C. & So, A.K. Identification of novel human T-cell receptor Vβ gene segments by the anchored-polymerase chain reaction. Immunogenetics 36, 314–318 (1992). https://doi.org/10.1007/BF00215660
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00215660