Analysis of cDNA clones specific for human T cells and the alpha and beta chains of the T-cell receptor heterodimer from a human T-cell line.

Abstract

We report the isolation and characterization of 19 classes of nonrearranging T cell-specific cDNA clones and two cDNA clones encoding the a and /3 chains of the T-cell antigen receptor from a human T-cell line, Jurkat. Results indicate that the human a-chain gene, like its /8-chain counterpart, undergoes somatic rearrangement in T cells. In addition, it shows sequence homology to its P-chain counterpart and immunoglobulin, indicating that the human a chain is also a member of the immunoglobulin supergene family. Sequence comparison suggests that the a chain also may be composed of variable (V), diversity (D), joining (J), and constant (C) region gene segments. The protein deduced from the cDNA sequence has a molecular weight of 29,995 and possesses six potential N-glycosylation sites. The availability of a-and /3-chain genes of the T-cell receptor from the same T-cell line provides tools to study their possible roles in recognition of antigens and major histocompatibility complex products by the human T-cell receptor.Antigen recognition by T lymphocytes is mediated by the T-cell antigen receptor composed of an a-and /3-chain heterodimer situated on the surface of these immunocompetent cells (1-3). About a year ago, we (4) and Hedrick et al.(5) cloned cDNAs that specified, respectively, the /3 chain of the human and mouse T-cell receptor (6). Subsequent studies have indicated that the genes encoding these structures undergo somatic rearrangement in T lymphocytes (7) and are homologous to the Ig genes (8). In addition, their genomic structures are similar, but not identical in organization, to Ig genes (9-13). These results indicate that genes encoding the T-cell receptor, at least the /8-chain gene, are different from Ig genes, though they most likely share a common ancestor (8). The finding that the human /3-chain variable (V) and constant (C) region genes (V and C) are located on chromosome 7 (14), where there are no Ig genes, confirmed that these genes are independent of Ig genes. These observations, coupled with the fact that a and ,B chains are approximately the same size, suggest that the a chain is probably also Ig-related and may contain similar V and C domains (8).In this manuscript, we report the analysis of a number of human T-cell-specific cDNA clones and describe the characterization and sequence of a human T-cell clone with homology to the a chain ofthe human T-cell antigen receptor. The results reveal that, like the /B-chain genes of the T-cell receptor (4, 5), the a-chain genes are homologous to Ig genes and undergo somatic rearrangement in T cells. While these studies were in progress, isolation of the murine a-chain cDNA (15, 16) and partial protein sequence of the human a chain (17, 18) were reported. The elucidation of the a and 83 chains of the T-cell receptor should provide the molecular tools necessary for the study of the recognition of antigens and major histocompatibility complex products by human T cells.

Non-T, non-B acute lymphocytic leukemias: Cellular origin based on molecular analyses of immunoglobulin and T-cell .ALPHA.- and .BETA.-chain receptor gene rearrangements.

“…4). The germ-line organization of the gene corresponding to pY 14 showed 7 germ-line bands in Eco RI-digested DNA, that is consistent to the result of Yanagi et al (1985). Eight out of 11 cases showed the germ-line arrangements.…”

“…The digests were electrophoresed through 0.75% agarose gels, transferred to nitrocellulose membranes, and hybridized with a 32P-labeled cDNA probe, as previously described in detail (Okabe et al 1987b). The plasmids containing human TcRf cDNAs were kindly provided by Dr. T. W. Mak ; a chain cDNA (pY 14) (Yanagi et al 1985) and constant region /3 chain cDNA (JUR-2) . JH probe was a 3.3 kb Eco RI-Hind III fragment of p (Ch4HIgu24) (Takahashi et al 1980) and Jx probe was a Sac I-Sac I fragment of embryonic Jx region gene (Hieter et al 1982), both obtained by the courtesy of Dr. T. Honjo.…”

A specific defect in CD3 gamma-chain gene transcription results in loss of T-cell receptor/CD3 expression late after human immunodeficiency virus infection of a CD4+ T-cell line.

“…The TCR a-and E-chain (TCR-a and TCR-f3) probes (23,24) were kindly provided by Tak Mak; the CD3-e probe (25), generated by polymerase chain reaction to the entire e-chain coding region, by Jason Loveridge (Institute for Molecular Medicine, Oxford, England); the CD3-; probe (26) by Allan Weissman; and the CD3-y probe (27) by M. J. Crumpton. The CD3-6 probe (28) Fig.…”

The T-cell receptor alpha, beta and gamma polymorphism in Japanese

“…The pY14 clone contains an insert of 1,101 pb that corresponds to the 5'-untranslated region, leader sequence, V, D, J and C regions and 3'-untranslated sequences (Yanagi et al, 1985).…”

“…By means of Southern blot analyses, twelve DNA samples were digested with 15 restriction endonucleases by using a probe that correspond to the human T-cell receptor a chain (Yanagi et al, 1985). Three out of the 15 enzymes presented polymorphic fragments.…”