© 1992 Oxford University Press

International Immunology, Vol. 4, No. 6, pp. 643 - 650

Analysis of J p gene segment usage by CD4 + and CD8 + human peripheral blood T lymphocytes Johan Grunewald, Mahmood Jeddi-Tehrani, Eva Pisa, Carl Harald Janson, Roland Andersson, and Hans Wigzell

Key words: human T cell receptor, skewed V gene usage, relative J gene usage, polymerase chain reaction

Abstract Certain T cell antigen receptor V gene products in man have been shown by us and others to display a reproducible bias for preferential expression in CD4 + or CD8 + T cell subsets. In order to investigate whether such a skewed representation of V gene segments is also present at the J gene segment level, we tested the relative J 0 gene usage by V0 5 . 1 + T cells, as this V^ gene is biased towards CD4+ T cell expression in virtually all individuals. To analyze the usage of the 13 Jp gene segments, we developed a new approach using V0 5.1 and C3 specific oligonucleotides as 5' and 3' primers respectively for polymerase chain reaction (PCR) amplification of cDNA derived from CD4 + or CD8 + peripheral blood lymphocyte (PBL) T cells. The PCR products were visualized for reactivity with individual J^ 1.1 - 1 . 6 and J 0 2 . 1 - 2 . 7 32P-labelled oligonucleotide probes using autoradiography and quantitative gel-scanning. Eleven normal blood donors provided the PBL T cells. The results showed that in every individual's V3 5 . 1 + T cell populations (CD4 and CD8), all v y j ^ combinations were used although at varying but reproducible levels for each Jp gene. Thus, no discernible disallowance of combinations existed. Moreover, we could show that six of 13 Je genes were unequally expressed when compared in pairs with regard to expression in CD4 + and CD8+ T cell subsets. The J 0 1.3, 1.4, 1.6, 2.5 and 2.6 gene segments showed a biased representation towards CD4+ T cells, while the J 0 2.7 gene segment was expressed more frequently by CD8+ T cells. We conclude that we have developed a simple and reproducible assay allowing the quantitative determination of individual J,, gene usage in relation to individual V genes in CD4/CD8 T cell subsets. In addition, one individual with an unusually high expression of peripheral Vfl 5 . 1 + CD4 + T cells (9.9%) showed an exceptionally high Je 2.1 gene segment usage. This finding indicates the sensitivity and usefulness of the experimental approach to detect even minor clonal expansions in complex cell populations.

Introduction The c*//3 T cell antigen receptor (TCR) is formed by the combination of the a and /? polypeptide chains. The genetic information for the variable region on each chain is generated by rearrangements of V (variable), D (diversity, only for the |3 chain) and J (joining) gene segments (1,2). In previous studies, we have been able to show a biased usage of some of the TCR V^ gene segment products towards the CD4 + T cell subpopulations, in peripheral blood as well as in thymus, and we found it to be most prominent for the V0 5.1 gene segment (3,4). Other groups have reported similar findings

on TCR V

Analysis of J beta gene segment usage by CD4+ and CD8+ human peripheral blood T lymphocytes.

Certain T cell antigen receptor V gene products in man have been shown by us and others to display a reproducible bias for preferential expression in ...
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