T-cell Receptor Vβ Gene Research Articles

Preferential but Not Exclusive T Cell Receptor Vβ Chain Utilization of Myelin Basic Protein and Peptide-Specific T Cell Clones in Mice

The repertoire of T cell receptor (TCR) Vβ chain utilization was investigated in PL/J, CXJ-1, SJL/J and B10.S → SJL/J chimeric mice in response to either myelin basic protein (MBP) or the strain-specific encephalitogenic peptide. Our analysis showed that there was an overlapping predominance in the TCR Vβ gene utilization in the MBP-specific responses, which were independent of the major histocompatibility complex (MHC) class II haplotype present, and the immunodominant peptide region recognized in these different strains. In those mice having the TCR Vβ b haplotype (PL/J, CXJ-1, and the B10.S → SJL chimera) either the TCR Vβ 4, 8, and 13 or the TCR Vβ 4, 6, and 13 predominated. In contrast, in mice with TCR Vβ a haplotype (SJL/J) Vβ 4, 6, and 17a were found. However, the quantitative distribution of these preferentially utilized TCR Vβ chains in each strain was defined by the MHC class II haplotype and the immunodominant peptide recognized. The expression of the Vβ8 gene product in the peripheral TCR repertoire did not always correlate with predominant Vβ8 utilization in the MBP-specific response.

Skewed distribution of TCR Vα 7-bearing T cells within tumor-infiltrating lymphocytes of HLA-A24(9)-positive patients with malignant glioma

The identification and propagation of T cells with anti-tumor reactivity is critical for understanding the human immune response to tumors, which may possibly be useful in the successful implementation of adoptive immunotherapy against cancer. In order to address this question, we examined the diversity of mRNA transcripts of T-cell receptor (TCR) Vα and Vβ genes in tumor-infiltrating lymphocytes (TIL) of 12 glioma specimens obtained at surgery. Using the polymerase chain reaction (PCR) method and primers for 18 different human TCR Vα and 22 Vβ families to analyze TCR V-(D)-J-C gene rearrangements, we detected a limited expression of TCR variable region, Vα genes and predominant usage of Vα 7 within glioma TIL. TCR Vβ gene usage was more diverse than that for Vα, but TCR Vβ 13.1 was dominantly expressed in 9 out of 12 patients. In addition, we analyzed the percentage of each Vα- and Vβ-bearing T-cell subpopulation in TIL as well as in peripheral blood lymphocytes (PBL) quantitatively. The distribution of T-cell subpopulation bearing each Vα or Vβ subfamily was variable and uneven in all cases. In 3 cases, the distribution of Vα 7-bearing T cells in TIL was far higher than in PBL. This phenomenon was not found in T cells bearing TCR Vβ 13.1. We also performed human leukocyte antigen (HLA) typing in these patients, and A24(9) was observed in 8 out of 11 patients. Among them all 3 patients who showed a skewed distribution of Vα 7-bearing T cells in TIL expressed HLA-A24(9). There was no correlation between particular class I or II type and TCR Vβ gene usage. From these results, it was strongly suggested that T cells bearing TCR Vα 7 might be targeted to antigenic determinants on glioma cells, and such T-cell population may be useful as effector cells for cancer immunotherapy.

Expression of T cell receptor Vβ transcripts in central nervous system of mice susceptible and resistant to Theiler's virus-induced demyelination

We utilized the polymerase chain reaction (PCR) to examine for preferential expression of T cell receptor (TcR) Vβs in T cells infiltrating the central nervous system (CNS) of mice infected with Theiler's virus. Infection of susceptible strains of mice with Theiler's virus results in demyelinating disease similar to multiple sclerosis. At 7 days following infection, no difference was observed in TcR Vβ usage in lymphocytes infiltrating the CNS between resistant (B10.K) and susceptible (B10.Q or SJL/J) mice. In susceptible mice with prominent demyelination (day 45 and 238 following infection), no preferential expression of TcR Vβs was observed in the CNS. There is strong evidence that T cells play a major role in pathogenesis of TMEV-induced demyelination. There is increasing evidence using recombinant inbred strain mice with TcR Vβ deletions that T cells expressing certain TcR Vβ genes may be critical in the disease process. Yet, analysis of TcR Vβ expression on T cells in CNS using PCR technology did not provide a way to dissect which antigen-specific T cells play a role in disease. These results confirm that during active demyelination specific as well as non-specific T cells are recruited to the CNS. Even though the pathogenesis of TMEV-induced disease may not be identical to that in multiple sclerosis, it is unlikely that similar approaches utilizing polymerase chain reaction will provide insights to the role of T cell receptors in the pathogenesis of multiple sclerosis.

Analysis of T Cell Receptor Vβ Regions Expressed by Rheumatoid Synovial T Lymphocytes

The T cell receptor (TCR) VII gene segment repertoire of T lymphocytes derived from peripheral blood of two healthy individuals and synovial tissue, synovial fluid and peripheral blood of three rheumatoid arthritis (RA) patients was analyzed. A sensitive assay based on the amplification of cDNA by the polymerase chain reaction (PCR) was used to analyze the levels of expression of 20 TCR Vβ gene segment families. The relative expression of Vβ gene segments in lymphocytes derived from peripheral blood, synovial tissue and synovial fluid was conserved over 155 days in one patient. Vβ9 transcripts were undetectable in the cells of this individual. In the two other patients the frequency of Vβ2 transcripts in synovial T cells of affected joints was significantly higher than in their peripheral blood lymphocytes. Dominance of distinct rearrangements among the Vβ2 transcripts from the synovial cells of these patients support the idea that the synovial T cell response is driven by antigen.

Limited heterogeneity of T-cell receptor Vβ gene expression in the early stage of insulitis in NOD mice

Non-obese-diabetic (NOD) mice spontaneously develop type I diabetes. The disease starts with T cells infiltrating the islets of Langerhans. We therefore examined the T-cell receptor (TCR) Vβ region repertoire in islet infiltrates from individual female NOD mice from 4 to 10 week old by polymerase chain reaction (PCR) using Vβ1-Vβ17 specific oligonucleotides. The study revealed a limited heterogeneity of TCR Vβ transcripts with a predominance of Vβ1 at the onset of insulitis, i.e. at 4 weeks of age. The TCR VDJβ sequences of the Vβ1 PCR fragments were identical in most of the individual mice. Among several different mice, similarities in the β-junctional regions were detected. In contrast, a large heterogeneity of TCR Vβ usage was found in mice with advanced insulitis, i.e., from 6 weeks of age on. Thus, these data suggest a limited heterogeneity of TCR Vβ usage with a predominance of Vβ1 at the initiation phase of the disease.

Patterns of T-Cell Receptor Variable β Gene Expression by Synovial Fluid and Peripheral Blood T-Cells in Rheumatoid Arthritis

Conflicting data have been reported regarding the presence or absence of a predominant variable (V) region T-cell receptor (TCR) gene in the peripheral blood or synovial fluid of patients with rheumatoid arthritis (RA). In this study we have used the polymerase chain reaction (PCR) to compare the level of TCR Vβ gene expression by peripheral blood mononuclear cells (PBMC) and by synovial fluid cells obtained from HLA DRB1 *0401 and *0404 RA patients. PCR was performed using cDNA synthesized from freshly obtained cells (not stimulated in vitro). The pattern of expression observed for most of the Vβ genes studied showed either preferential expression by PBMC or similar levels of expression between PBMC and synovial fluid T-cells. However, among individual patients ( N = 5), several Vβ genes were identified that were expressed to a significantly greater degree by synovial fluid cells. Vβ14 expression was detected in PBMC of all patients and the level of transcripts encoding Vβ14 increased following stimulation with immobilized anti-CD3 and IL-2. In vitro manipulation of populations of T-cells was found to alter the level of expressed Vβ gene products. A Vβ gene common to all patients that was consistently deleted from PBMC or expressed to a greater degree by resident, unsorted synovial fluid cells compared to PBMC was not identified.

Susceptibility for multiple sclerosis is determined, in part, by inheritance of a 175-kb region of the TcR Vβ chain locus and HLA class II genes

Genetic makeup thought to affect susceptibility to multiple sclerosis (MS) and current evidence suggests that multiple genes may be involved. We have mapped a potential susceptibility gene or genes in the germ-line T cell receptor (TcR) Vβ region of multiple sclerosis (MS) patients. Six restriction fragment length polymorphisms (RFLPs) spanning approximately 600 kb of the TcR Vβ region were used to define TcR haplotypes in 197 Caucasian controls and 83 Caucasian MS patients in the chronic progressive stage of the disease. The distribution of TcR subhaplotype frequencies was significantly different only in the approx. 175-kb region between RFLPs defined by Vβ8.1 and Vβ11. Stratification of the MS patients into HLA-DR2 + ( n = 51) and HLA-DR2 − ( n = 32) populations demonstrated that the subhaplotype frequencies differed from the control population significantly only in the HLA-DR2 + (corrected P = 0.00007) and not in the HLA-DR2 − (corrected P = 0.46) population. Subhaplotypes which are rare in the normal population are overrepresented in the HLA-DR2 + MS patient population and confer a relative risk of 4.06. These results indicate the existence of an MS susceptibility gene within the TcR Vβ region, and provide new evidence for gene complementation between a HLA class II gene and TcR Vβ gene(s) in conferring susceptibility to MS.

HIV infection leads to differential expression of T-cell receptor V beta genes in CD4+ and CD8+ T cells.

To analyse variation in T-cell receptor (TCR) V beta gene expression in T cells in HIV-infected individuals. Because there are very few monoclonal antibodies available for studying TCR V beta gene expression, we used polymerase chain reaction (PCR) to analyse the TCR V beta repertoire in HIV-infected individuals using specific primers for 20 distinct families of TCR V beta. Evaluation of TCR V beta gene expression in peripheral blood from HIV-1-infected individuals [two in Centers for Disease Control (CDC) stage II, five in CDC stage III and four in CDC stage IV]. Complementary DNA was produced from CD4+ and CD8+ T cells, amplified by PCR and analysed after Southern blotting and hybridization with a C beta-specific oligonucleotide probe. V beta gene expression was dramatically modified, especially in AIDS patients. The CD4+ T-cell subset showed both overexpression (V beta 2) and deletions or underexpression (V beta 9-V beta 20), whereas these gene segments were expressed normally in the CD8+ subset. Only V beta 3 was deleted or underexpression in both CD4+ and CD8+ populations in AIDS patients. HIV-1 infection induces CD4+ T-cell deficiency, both in total numbers and by causing a paucity of select V beta gene expression in this subset. In addition, the V beta 3 gene family was deleted or underexpressed was observed in both CD4+ and CD8+ T-cell subsets from patients in CDC stage IV. These results are compatible with changes in V beta gene expression known to occur under the action of endogenous or exogenous superantigens.

A single universal primer for the T-Cell receptor (TCR) variable genes enables enzymatic amplification and direct sequencing of TCRβ cDNA of various T-cell clones

We designed a primer for the PCR directed against a highly conserved sequence of the TCR Vβ gene. The Vβ-universal primer, in combination with a constant region-specific primer, enabled us to amplify TCRβ cDNA of allo-HLA class-II-reactive T-cell clones by PCR without prior knowledge of their Vβ sequences. The amplified TCR cDNA was purified by agarose gel electrophoresis and subjected to direct sequencing. In nine of ten T-cell clones analyzed, direct TCR sequencing gave readable sequence ladders, including two-thirds of Vβ, junctional, and Jβ regions. One T-cell clone gave an unreadable mixed-profile sequence ladder, indicating that this clone expressed more than one major TCRβ transcript. Even in this case, however, it was possible to determine two different TCRβ sequences separately using sequence primers specific to one of the 13 Jβ segments deduced from the mixed ladder. Thus, direct sequencing utilizing the single Vβ-universal primer enabled a simple, rapid, and reliable sequence determination of TCRβ cDNA of all T-cell clones analyzed.

Analysis of the TCR Vβ Specificities of Bacterial Superantigens Using PCR

Several bacterial proteins have been recently identified as superantigens which stimulate large numbers of T cells expressing particular T-cell receptor (TCR) Vβ elements. This suggests important roles for these proteins in the pathogenesis of bacterial infections. However, before the superantigenic property can be assigned to a particular protein, the purity needs to be carefully evaluated and several measures need to be taken to rule out the presence of minute amounts of contamination by other superantigens such as bacterial toxins. Here we describe a technique that can be used effectively to address this problem. This technique exploits the fact that superantigens have a signature Vβ specificity and, therefore, analysis by PCR of TCR Vβ gene expression which shows the Vβ expansion profile can be used to determine the superantigenicity of a protein and to detect cross-contamination by other superantigens. 1993 Academic

Human T-cell receptor Vβ gene segments involved in an insertion/deletion-related polymorphism: T.M. Zhao, S.E. Whitaker, and [formula omitted], Laboratory of Immunogenetics, NIAID, NIH, Rockville, MD

Human T-cell receptor Vβ gene segments involved in an insertion/deletion-related polymorphism: T.M. Zhao, S.E. Whitaker, and [formula omitted], Laboratory of Immunogenetics, NIAID, NIH, Rockville, MD

Usage of human T-cell receptor Vβ, Jβ, Cβ, and Vα gene segments is not proportional to gene number

Certain T-cell receptor (TCT) β-chain variable (V), joining (J), and constant (C) gene segments, as well as TCRα-chain V gene segments, are disproportionally represented in TCR α and β cDNA libraries derived from PHA-stimulated peripheral blood lymphocytes. Sequences of 138 TCRα clones and 96 TCRβ clones were determined and of these 128 TCRα clones and 88 TCRβ clones were found to contain unique combinations of V, J, and C gene segments or to display diversity in N region nucleotides. The frequency of the V, J, and C genes used in the assembly of unique transcripts was ascertained. Of the 24 reported Vβ genes, families, 21 were observed among the 88 TCRβ clones including four Vβ families (Vβ1, Vβ2, Vβ3, and Vβ4) that were represented in the sample 2 1 2 −5 times more frequently than would be expected on the basis of copy number within the gene complex. Seventy-eight percent of the clones were positive for Cβ2 and more than half of the clones (53%) used one of two Jβ2 genes: Jβ2.1 was present in 27 clones and Jβ2.7 in 20 clones. TCR Vα families were also disproportionately represented in this sample. Twenty-five of 30 Vα families were observed in the sample of 128 clones including six recently reported Vα families. Three Vα families, Vα2, Vα8, and Vα23, accounted for ≈40% of the TCRα clones and were represented at 18%, 9.4%, and 13.3%, respectively. Both Vα2 and Vα8 gene families contain more than one gene; thus the number of clones observed in these families may, in part, be related to gene number. However, Vα23, which appears to be a single-copy gene family, is significantly overrepresented in this sample. Although disproportional usage of Vβ genes may be accounted for by superantigen exposure, reasons for disproportional usage of Jβ, Cβ, and Vα genes are presently unknown.

Secretion of T cell receptor fragments from recombinant Escherichia coli cells

This study describes the secretion and purification of T cell receptor (TCR) Vα, Vβ domains and single chain Vα-Vβ fragments (scTCRs) from recombinant Escherichia coli cells. The TCR Vα and Vβ genes are derived from a T cell hybridoma that is associated with disease pathogenesis in murine experimental allergic encephalomyelitis (EAE). Circular dichroism (c.d.) analyses of the single domains and the scTCR indicate that they are folded into β-pleated sheet structures similar to those of immunoglobulin variable domains. The secreted TCR fragments can be purified in milligram quantities, and could therefore be used in high-resolution structural studies, in immunization to generate anti-clonotypic antibodies or in vaccination.

Multiple TCR V beta usage by infiltrates of young NOD mouse islets of Langerhans. A polymerase chain reaction analysis.

Because a restricted repertoire of T-cell receptor (TCR) V beta gene expression has been reported in other autoimmune diseases, the possibility of similarly restricted V beta gene expression by T-cell infiltrates of NOD mouse islets was examined. With isolated islets from 4- to 12-wk-old NOD mice, a prospective polymerase chain reaction analysis with 18 V beta-specific oligonucleotide primers was performed on the noncloned and unexpanded islet-infiltrating T cells. The methodology used permitted the detection of a minimum of 50 T cells. In contrast to the restricted TCR V beta gene usage reported for other autoimmune diseases, infiltrates of even the youngest mice were characterized by expression of multiple V beta gene segments.

T-cell receptor Vbeta gene usage by lymphocytes infiltrating human renal allografts.

T cell lines have been derived from human kidney allograft biopsies using mitogenic stimulation. Southern blotting using a T-cell receptor (TCR) Cbeta probe revealed an oligoclonal pattern of rearranged bands in all 12 samples analysed. In some cases, differences in band patterns were noted between independent cultures from the same biopsy. Most T-cell clones derived from 2 biopsies showed different patterns of rearranged bands. The polymerase chain reaction (PCR) was used to study TCR Vbeta gene usage in allograft-derived T-cell cultures. This was more sensitive and more informative than Southern blotting and revealed that most TCR Vbeta genes were expressed in T cells from biopsies showing cellular rejection. The potential usefulness of this technique to quantify TCR V gene usage in allospecific T-cell populations is discussed.

T-cell receptor Vβ gene usage by lymphocytes infiltrating human renal allografts

T-cell receptor Vβ gene usage by lymphocytes infiltrating human renal allografts

Limited heterogeneity of autoantigens and T cells in autoimmune diseases?

For many induced and spontaneous autoimmune diseases, a predominant role for T cells in the organ-specific destruction process has been shown. In one of the induced models of autoimmunity, experimental allergic encephalomyelities (EAE), a very small heterogeneity of T-cell receptor (TcR) molecules is expressed by the pathogenic T cells in both rats and mice. Contrary to induced autoimmune diseases, little is known about the autoantigens recognized by these autoimmune T cells and the heterogeneity of their TcR in spontaneous autoimmune diseases. The aim of this work was to establish a system which allows characterization of relevant autoantigens in spontaneous insulin-dependent diabetes mellitus (IDDM) in non-obese diabetic (NOD) mice. A completely different approach was taken to characterize the gene products of the minor lymphocyte stimulatory (Mls) loci. These gene products are responsible for the clonal elimination or the clonal stimulation of T cells expressing particular TcR Vβ genes and therefore could be implicated in induction of autoimmune diseases by oligoclonal T-cell populations. The finding that Mls antigens are encoded by retroviral sequences leads to the hypothesis that viruses could be the inducing agents of autoimnune diseases.

Germline sequence of two human T-cell receptor V beta genes: V beta 8.1 is transcribed from a TATA-box promoter.

Journal Article Germline sequence of two human T-cell receptor Vβ genes: Vβ8.1 is transcribed from a TATA-box promoter Get access W.J. Smith, W.J. Smith Medical Research Council Laboratory of Molecular BiologyHills Road, Cambridge CB2 2QH, UK 1 Present address: Department of Histopathology, University College Medical School, University Street, London WC1E 6JJ, UK Search for other works by this author on: Oxford Academic PubMed Google Scholar A. Tunnacliffe, A. Tunnacliffe Medical Research Council Laboratory of Molecular BiologyHills Road, Cambridge CB2 2QH, UK Search for other works by this author on: Oxford Academic PubMed Google Scholar T.H. Rabbitts T.H. Rabbitts Medical Research Council Laboratory of Molecular BiologyHills Road, Cambridge CB2 2QH, UK Search for other works by this author on: Oxford Academic PubMed Google Scholar Nucleic Acids Research, Volume 15, Issue 12, 25 June 1987, Page 4991, https://doi.org/10.1093/nar/15.12.4991 Published: 25 June 1987

Molecular genetics of murine lupus.

Recent molecular analyses have suggested the following. The Ig VH genes of lupus mice are essentially normal or, alternatively, germ line genes encoding autoantibodies are present in both autoimmune and normal mice. Lupus-associated autoantibodies derive from a relatively large number of clonally unrelated B cells and are encoded by similar germ line elements to those encoding antibodies to foreign antigens without preferential expression of any particular VH–VL gene family or DH–VL elements. The genomic organization of T cell receptor (TCR) Cα-Cβ genes is similar in lupus and normal mice, except for a deletion encompassing Cβ1–Dβ2–Jβ2 elements unique to NZW mice. Most TCR Vβ genes are present in the genomes of lupus mice. The T cell receptor Vβ genes expressed in enlarged lymph nodes of MRL-lpr/lpr lupus mice are diverse, but show a preferential expression of the Vβ8.2/8.3 genes.