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A Tunnacliffe

Publications and source records attributed to A Tunnacliffe.

At least 55 records · Page 3Linked to original sources
PubMed1990

DNase I-defined chromatin configuration of the human CD3 gene cluster.

The three CD3 genes on human chromosome 11q23 encode proteins (gamma, delta and epsilon) which form part of the antigen receptor on T lymphocytes. All three genes are clustered within 50 kb and are activated approximately contemporaneously during the early stages of T cell ontogeny. In order to pinpoint potential regulatory sequences important for locus activation and tissue-specific gene expression, the chromatin structure of almost 90 kb of this region has been probed in five cell lines using the endonuclease pancreatic DNase I. A set of DNase I hypersensitive (HS) sites has been defined in T cell chromatin, five of which were strong and not found in non-T cells, with the exception of the erythroleukaemia cell line K562, in which three sites were weakly expressed, correlating with a low level of delta mRNA. The subset of five HS sites map close to the CD3 genes and lie in regions which contain elements of defined function: the gamma promoter; the delta promoter and its 3' enhancer; and the epsilon promoter and its 3' enhancer. Since no further major T cell-restricted HS sites lie within the 90kb of the CD3 locus analysed, these five regions may contain all the sequences important for CD3 gene expression.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1990

T cell activation by a bispecific anti-CD3/anti-major histocompatibility complex class I antibody.

A bispecific anti-CD3/anti-major histocompatibility complex (MHC) class I antibody (Ab) that is able to activate human T cells in the absence of a second signal has been used to compare activation by this Ab to signaling by anti-CD3 Ab or by antigen (Ag). We have studied early, intermediate and late events that occur after triggering of a tetanus toxoid-specific T cell clone. While bivalent anti-CD3 Ab induce transient (less than 20 min) intracellular Ca2+ concentration increases, the bispecific Ab, like antigen-presenting cells plus Ag, produces sustained (greater than 2 h) elevated intracellular Ca2+ concentration. In addition, while the anti-CD3 Ab only induces low levels of c-myc mRNA lasting less than 3 h, the bispecific Ab induces high levels which are maintained for at least 8 h after triggering. Late events, such as interleukin 2 receptor expression and proliferation, occurring more than 20 h after activation, were seen only when T cells were stimulated by bispecific Ab or by antigen-presenting cell and Ag. Cells activated by linking T cell receptor (TcR)/CD3 to MHC class I antigens could not be activated by cross-linking the MHC and TcR/CD3 structures separately. Therefore, we propose that the bispecific Ab functions by modifying the membrane dynamics of the TcR/CD3 complex, rather than by the generation of a second signal through class I antigens.

Antigen-Presenting Cells↗
PubMed1990

Characterization of the CD3 gamma and delta invariant subunits of the sheep T cell antigen receptor.

The CD3 gamma and delta chains of the sheep T cell antigen receptor have been characterized by cDNA cloning. In comparison to human and mouse protein sequences, short oligopeptide stretches have been deleted from both the gamma and delta chains. However, apart from further variability in the number and position of potential N-linked glycosylation sites, the general structure of the CD3 gamma and delta proteins is markedly conserved. Particularly remarkable are two motifs consisting of 8 and 32 amino acids, respectively, which locate in the C-terminal half of each protein and which have been precisely conserved in both the CD3 gamma and delta chains of all three species. These motifs probably contribute structural properties essential for the function of the CD3/T cell receptor complex. mRNA of the sheep CD3 gamma (1.5 and 1.2 kb) and CD3 delta (1.5 kb) chains are transcribed from closely linked, single-copy genes, suggesting that common mechanisms for the regulation of CD3 genes are also preserved between species.

Amino Acid Sequence↗
PubMed1990

A physical linkage group in human chromosome band 11q23 covering a region implicated in leukocyte neoplasia.

Six genes on human chromosome band 11q23 have been linked by pulse-field gel electrophoretic analysis with the order cen-CD3E-CD3D-CD3G-PBGD-CBL2-THY1-qter. The corresponding long-range restriction map covers 1.8 Mb, the telomeric half of which (PBGD-CBL2-THY1) is localized to subband 11q23.3. Four genes (CD3E, CD3D, CD3G, and PBGD) can be positioned precisely, and with known transcriptional orientation, with respect to rare-cutter restriction sites. The linkage group covers a region implicated in leukocyte malignancy: several nonrandom neoplasia-associated translocation chromosomes have breakpoints which separate the CD3 genes from PBGD, CBL2, and THY1. Thus, we are able to localize such breakpoints, and consequently any affected candidate genes, to the 750 kb between CD3G and PBGD.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1990

Physical mapping of the CD3 gene cluster.

By a combination of pulsed-field gel electrophoresis and molecular cloning techniques, the human CD3-gamma, CD3-delta and CD3-epsilon genes have been shown to be clustered in approximately 50 kb of chromosome 11q23. The mouse genes are probably organised in a similar fashion on chromosome 9. An approach to the construction of a long-range physical map surrounding the CD3 gene cluster is described, which will complement the genetic maps of the region.

Animals↗
PubMed1989

Production and secretion of recombinant soluble CD3 polypeptides by myeloma-derived transfectant clones.

Soluble forms of three human CD3 proteins have been produced by recombinant DNA techniques. The extracellular domain of CD3-gamma, -delta or -epsilon has been linked to the constant region of mouse immunoglobulin kappa light chain to form gamma-kappa, delta-kappa and epsilon-kappa chimaeric proteins. These are secreted by mouse myeloma-derived transfectant cell lines and are immunoprecipitable by CD3- or kappa-specific polyclonal antisera. Yields of 100-500 micrograms secreted recombinant proteins per litre of culture medium were obtained, which could be purified by anti-kappa affinity chromatography. The production of soluble CD3 illustrates the applicability of this technology to a loosely associated protein complex.

Antigens, Differentiation↗
PubMed1989

The majority of human CD3 epitopes are conferred by the epsilon chain.

Transgenic mouse T cells expressing the human CD3 epsilon chain bind the majority (29/36) of monoclonal antibodies (mAbs) specific for human CD3. A proportion of these mAbs are also able to recognize isolated CD3 epsilon in a soluble, recombinant form. Thus, CD3 epsilon can confer most CD3 epitopes on the TCR--CD3 complex, but many determinants may require assembly of the complex for their formation. A number of mAbs did not recognize epsilon-transgenic T cells and probably need other CD3 subunits for binding. CD3-specific mAbs from each of the three groups defined here, as well as mAbs directed against the TCR alpha beta heterodimer, are all able to activate T cells. Therefore mAb attachment at several different sites on the TCR--CD3 complex can give rise to activation signals. This suggests that the cross-linking function of mitogenic antibodies may be their most significant property, rather than the perturbation of a particular 'functional epitope'.

Animals↗
PubMed1988

Thymocyte clones from 14-day mouse embryos. II. Transcription of T3 gamma gene may precede rearrangement of TcR delta and expression of T3 delta, T3 epsilon and T11 genes.

We have studied the state of TcR delta gene and the expression of T3 delta, T3 epsilon, T3 gamma and T11 (CD2) genes in the fetal thymocyte (FT) clones A2, G15, H5, E10, D5, H12, F1 and D11 obtained from a 14-day B10.BR mouse fetal thymus. The eight FT clones contain the TcR delta gene in the germ-line configuration as determined by Southern blot analysis. With the exception of clones E10 and D5, the other six FT clones express normal sized transcripts for T3 gamma gene and none of the eight FT clones produced detectable RNA transcripts for T3 delta, T3 epsilon and T11 genes as assessed by Northern blot analysis. Together with our previous work showing that all eight FT clones contain the TcR gamma and the TcR beta gene clusters in the germ-line configuration, the data indicate that the FT clones represent the earliest stage of T cell development identified within the thymus. Our results provide evidence that (a) the T3 gamma gene is the first of the genes that encode components of a TcR/T3 complex to be expressed in ontogeny within the thymus; (b) the T3 (delta, epsilon, gamma) genes are switched on asynchronously and their expression must be differentially regulated, and (c) the T11 gene product may not be involved in early stages of T cell development within the thymus.

Animals↗
PubMed1988

Organization of the human CD3 locus on chromosome 11.

Three CD3 genes, gamma, delta, epsilon, map to human chromosome 11 band q23. A 75-kb region of 11q23 containing all three genes has been characterized by field inversion gel electrophoresis and molecular cloning. The CD3 genes lie within a stretch of 50 kb of DNA, orientated 3'-CD3 gamma-5': 5'-CD3 delta-3': 3'-CD3 epsilon-5'. Only 1.6 kb separate the CD3 gamma and CD3 delta genes, while the CD3 delta and CD3 epsilon genes are approximately 22 kb apart. CD3 gene expression is one of the first events during the maturation of the thymocyte, and therefore the clustering of CD3 genes may define a chromatin domain which is activated early in T cell development.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1987

Physical linkage of three CD3 genes on human chromosome 11.

T-cell antigen receptors are associated on T cell surfaces with a complex of proteins called CD3 (formerly T3). Human CD3 consists of at least four proteins, gamma, delta, epsilon and zeta, and all but the latter have been cloned as cDNA. Using standard cloning techniques, together with field inversion gel electrophoresis, we have demonstrated the physical linkage of three CD3 genes. The genes for CD3 gamma and CD3 delta are situated close together, about 1.6 kb apart, organized in a head-to-head orientation. The gene encoding CD3 gamma has been sequenced, and is split into seven exons spread over 9 kb of DNA. Like CD3 delta, CD3 gamma gene has an unusual promoter which lacks a TATA-box and potential Sp1 binding sites. The CD3 gamma-CD3 delta gene pair is within 300 kb of the CD3 epsilon gene, and therefore these genes form a tightly linked cluster in chromosome 11 band q23. The clustering of the CD3 genes may be significant in terms of their simultaneous activation during T-cell development.

Amino Acid Sequence↗
PubMed1986

T3 delta pre-mRNA is transcribed from a non-TATA promoter and is alternatively spliced in human T cells.

The human T-cell antigen receptor-associated T3 complex consists of at least three polypeptides, gamma, delta and epsilon. cDNA clones for the delta-chain have recently been obtained and we have used such clones to isolate the T3 delta gene. The gene has been sequenced and comprises five exons, spread over approximately 3.7 kb of DNA. Transcription of the T3 delta gene is initiated from a non-TATA promoter. S1 mapping experiments and the sequence of a novel cDNA clone show that T3 delta mRNA exists in two forms in T cells. Alternative splicing of pre-mRNA sequences corresponding to the third exon of the T3 delta gene accounts for the two species of mRNA. A putative protein, produced by translation of the shorter mRNA, would lack a transmembrane region and might be secreted or associated with the outer surface of the cell.

Amino Acid Sequence↗
PubMed1985

Sequence of the D beta 2-J beta 2 region of the human T-cell receptor beta-chain locus.

We have sequenced the region encompassing a D beta 2 segment and the J beta 2 segments of the human T-cell antigen receptor beta-chain genes. The D beta 2 element lies about 650 base pairs upstream of a cluster of seven potentially functional J beta 2 sequences and one J beta 2 pseudogene. Examination of human beta-chain cDNA sequences which involve rearranged D beta 2 and J beta 2 elements demonstrates that N-region, as well as junctional, diversity can occur during D-J joining. Further, we present evidence for possible somatic mutation in active J beta 2 segments.

Amino Acid Sequence↗
PubMed1985

Sequence and evolution of the human T-cell antigen receptor beta-chain genes.

We present the nucleotide sequences of the two genomic constant (C)-region gene segments, C beta 1 and C beta 2, encoding the beta chain of the human T-cell antigen receptor. The two C beta genes are organized identically to each other and to the corresponding mouse genes, both having four exons, whose boundaries were confirmed from the sequence of a C beta 2 cDNA clone from the T-cell line MOLT-4. The predicted amino acid sequences of human C beta 1 and C beta 2 differ at only five positions, which suggests that the proteins have very similar functions. This similarity is the result of strong nucleotide-sequence conservation in protein-coding regions, which extends to silent positions. A quantitative analysis of an alignment of the nucleotide sequences of the two human genes shows that whereas the 5' ends (including the first exon) are extremely homologous, the 3' ends are widely divergent, with other regions having intermediate levels of homology. Analysis of published data [Gascoigne, N.R.J., Chien, Y., Becker, D.M., Kavaler, J. & Davis, M.M. (1984) Nature (London) 310, 387-391] shows that the mouse C beta 1 and C beta 2 genes are also virtually identical in their first exons but more divergent in the remaining coding regions. Therefore, partial gene conversion events may have occurred during the evolution of both human and mouse C beta genes.

Amino Acid Sequence↗
PubMed1984

Human muscle cell surface antigen 16.3A5 is encoded by a gene on chromosome 11.

A monoclonal antibody (McAb), 16.3A5, has been characterized that reacts with a cell surface antigen of human muscle cells and a variety of nonmuscle cells. The gene controlling synthesis of 16.3A5 antigen has been assigned to human chromosome 11 by assessing McAb reactivity on a panel of mouse-human cell hybrids. The 16.3A5 has a novel specificity distinct from other chromosome 11-encoded antigens such as W6/34, F10.44.2, TRA1.10, and 4D12 antigens.

Adult↗
PubMed1984

Relationships between genes on human chromosome 11 encoding cell-surface antigens.

Genes encoding seven monoclonal antibody-defined cell-surface antigens have been regionally mapped on human chromosome 11, and compared to those of the AL complex defined by polyclonal antibodies using mutational analysis. MIC1, encoding W6/34 antigen, is probably identical to S1, previously mapped to 11pter-p13. MDU1 and MIC8, encoding 4F2 and TRA-1.10 antigens, respectively, are probably identical to S2(a4) and map to 11q13-q22. MIC9, which governs expression of 4D12 and 2E2 antigens, and maps to 11q22-qter, is not related to any of the five AL genes. MIC4 and MIC11, both mapping to 11pter-p13, may have some relationship to S3 and S1, respectively, but identity has not been proven.

Antibodies, Monoclonal↗
PubMed1984

Expression of simian virus 40 large T antigen in embryonal carcinoma cell hybrids.

Previous work has shown that murine embryonal carcinoma cells are refractory to infection with various viruses, including simian virus 40. Thus, large T and small t antigens, the products of the simian virus 40 early region, are not produced when the virus infects embryonal carcinoma cells, in contrast to other cell types. We show, by qualitative and quantitative analyses, that embryonal carcinoma cell hybrids, containing a simian virus 40 early region integrated into human DNA, are capable of producing viral large T antigen.

Animals↗