Conserved CDR3 Regions in T-Cell Receptor (TCR) CD8+T Cells That Recognize the Tax11-19/HLA-A*0201 Complex in a Subject Infected with Human T-Cell Leukemia Virus Type 1: Relationship of T-Cell Fine Specificity and Major Histocompatibility Complex/Peptide/TCR Crystal Structure

ABSTRACT We investigated the T-cell receptor (TCR) repertoire of CD8+ T cells that recognize the Tax11-19 immunodominant epitope of Tax protein expressed by human T-cell leukemia virus (HTLV-1) that is implicated in the disease HTLV-1-associated myelopathy (HAM/TSP). A panel of Tax11-19-reactive CD8+ T-cell clones was generated by single-cell cloning of Tax11-19/HLA-A*0201 tetramer-positive peripheral blood lymphocytes from an HTLV-1-infected individual. The analyses of TCR usage revealed that the combination of diverse TCR alpha and beta chains could be used for the recognition of Tax11-19 but the major population of T-cell clones (15 of 24 clones) expressed the TCR V beta 13S1 and V alpha 17 chain. We found striking similarities in CDR3 regions of TCR alpha and beta chains between our major group of CD8+ T-cell clones and those originating from different subjects as previously reported, including TCRs with resolved crystal structures. A 3-amino-acid sequence (PG-G) in the CDR3 region of the V beta chain was conserved among all the Tax11-19-reactive T-cell clones expressing V beta 13S1 and V alpha 17 chains. Conserved amino acids in the CDR3 region do not directly contact the Tax11-19 peptide, as corroborated by the crystal structure of B7-TCR, a TCR that is almost identical to VB13S1 clones isolated in this study. Analysis of fine peptide specificity using altered peptide ligands (APL) of Tax11-19 revealed a similar recognition pattern among this panel of T-cell clones. These data suggest that the PG-G amino acids in the CDR3 beta loop provide a structural framework necessary for the maintenance of the tertiary TCR structure.

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Modulation of promiscuous T cell receptor recognition by mutagenesis of CDR2 residues.

Journal of Experimental Medicine ◽

10.1084/jem.183.5.2043 ◽

1996 ◽

Vol 183 (5) ◽

pp. 2043-2051 ◽

Cited By ~ 22

Author(s):

J V Brawley ◽

P Concannon

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Jurkat Cells ◽

Single Amino Acid ◽

Peptide Antigen ◽

T Cell Clones ◽

Human T Cell ◽

Cdr3 Region ◽

Cell Clones

The T cell receptor (TCR) recognizes a ligand composed of a major histocompatibility complex (MHC) molecule and a peptide antigen. Prior studies of murine T cell clones have demonstrated that residues in the CDR3 region of TCR interact with amino acids in the peptide during MHC-restricted antigen recognition. However, the questions of whether direct TCR MHC contacts are made and where such contact sites might map in the TCR have not been resolved. In this study, we have taken advantage of the promiscuous recognition of a peptide from influenza virus (HA 307-319) by human T cell clones to map sites in the TCR that mediate differences in human leukocyte antigen-D related (HLA-DR) restriction in the presence of a common peptide antigen. Site-specific mutagenesis of cloned TCR genes and transfection into Jurkat cells were used to demonstrate that single amino acid substitutions in CDR2 of the TCR-alpha chain controlled whether a T cell was restricted by the product of a single DR allele (DR7) or would respond to the HA 307-319 peptide when presented by the products of one of several different DR alleles (DR1, DR4, DR5, or DR7). Because the relevant DR alleles are defined by polymorphism in the DR-beta chain, these results also suggest a rotational orientation for recognition in which TCR-alpha interacts with DR beta.

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Human T-Cell Leukemia Virus Type 1 Tax Protein Down-Regulates Pre-T-Cell Receptor Alpha Gene Transcription in Human Immature Thymocytes

Journal of Virology ◽

10.1128/jvi.00766-06 ◽

2006 ◽

Vol 81 (1) ◽

pp. 301-308 ◽

Cited By ~ 6

Author(s):

Mélanie Wencker ◽

Céline Sausse ◽

David Derse ◽

Louis Gazzolo ◽

Madeleine Duc Dodon

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Gene Transcription ◽

Leukemia Virus ◽

Cell Receptor ◽

Cell Leukemia ◽

Cell Leukemia Virus Type ◽

Human T Cell ◽

T Cell Leukemia Virus

ABSTRACT The human pre-T-cell receptor alpha (TCRα; pTα) gene encodes a polypeptide which associates with the TCRβ chain and CD3 molecules to form the pre-TCR complex. The surface expression of the pre-TCR is pTα dependent, and signaling through this complex triggers an early αβ T-cell developmental checkpoint inside the thymus, known as β-selection. E2A transcription factors, which are involved at multiple stages of T-cell development, regulate the transcription of the pTα gene. Here we show that the regulatory protein Tax of the human T-cell leukemia virus type 1 (HTLV-1) efficiently suppresses the E47-mediated activation of the pTα promoter. Furthermore, we report that in Tax lentivirally transduced human MOLT-4 T cells, which constitutively express the pTα gene, the amount of pTα transcripts decreases. Such a decrease is not observed in MOLT-4 cells transduced by a vector encoding the Tax mutant K88A, which is unable to interact with p300. These data underline that Tax inhibits pTα transcription by recruiting this coactivator. Finally, we show that the expression of Tax in human immature thymocytes results in a decrease of pTα gene transcription but does not modify the level of E47 transcripts. These observations indicate that Tax, by silencing E proteins, down-regulates pTα gene transcription during early thymocyte development. They further provide evidence that Tax can interfere with an important checkpoint during T-cell differentiation in the thymus.

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T-Cell Receptor/CD28 Engagement When Combined with Prostaglandin E2 Treatment Leads to Potent Activation of Human T-Cell Leukemia Virus Type 1

Journal of Virology ◽

10.1128/jvi.77.20.11170-11179.2003 ◽

2003 ◽

Vol 77 (20) ◽

pp. 11170-11179 ◽

Cited By ~ 11

Author(s):

Nancy Dumais ◽

Marie-Ève Paré ◽

Simon Mercier ◽

Salim Bounou ◽

Susan J. Marriot ◽

...

Keyword(s):

Gene Expression ◽

T Cell ◽

T Cell Receptor ◽

Leukemia Virus ◽

Cell Receptor ◽

Cell Leukemia ◽

Cell Leukemia Virus Type ◽

Human T Cell ◽

T Cell Leukemia Virus

ABSTRACT Infection with human T-cell leukemia virus type 1 (HTLV-1) is characterized by long latency periods, indicating that viral gene expression is under tight control. There is presently little information available regarding the nature of extracellular stimuli that can transactivate the regulatory elements of HTLV-1 (i.e., long terminal repeat [LTR]). To gain insight into the biological importance of externally induced activation pathways in virus gene expression, primary and established T cells were transfected with HTLV-1-based reporter gene vectors and then were treated with agents that cross-linked the T-cell receptor (TCR) or the costimulatory CD28 molecule with prostaglandin E2 (PGE2). We demonstrated that a potent induction of HTLV-1 LTR-driven reporter gene activity was seen only when the three agents were used in combination. Interestingly, similar observations were made when using C91/PL, a cell line that carries integrated HTLV-1 proviral DNA. This TCR-CD28-PGE2-mediated increase in virus transcription was dependent on protein kinase A activation and induction of the cAMP response element binding protein. Experiments with a mutated reporter construct further revealed the importance of the Tax-responsive elements in the HTLV-1 LTR in the observed up regulation of virus gene expression when TCR/CD28 engagement was combined with PGE2 treatment. The protein tyrosine kinases p56 lck and the transmembrane tyrosine phosphatase CD45 were all found to be involved in TCR-CD28-PGE2-directed increase in HTLV-1 LTR activity. This study presents new information on the possible mechanisms underlying reactivation of this retrovirus.

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Human T-cell leukemia virus type I(HTLV-I) infection of T cells bearing T-cell receptor γδ: Effects of HTLV-I infection on cytotoxicity

International Journal of Cancer ◽

10.1002/ijc.2910500318 ◽

1992 ◽

Vol 50 (3) ◽

pp. 431-437 ◽

Cited By ~ 3

Author(s):

Masaki Yasukawa ◽

Akira Inatsuki ◽

Yoshihiro Yakushijin ◽

Yuzuru Kobayashi

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Receptor ◽

Leukemia Virus ◽

Cell Receptor ◽

Type I ◽

Cell Leukemia ◽

Cell Leukemia Virus Type ◽

Human T Cell ◽

T Cell Leukemia Virus

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Variable Immortalizing Potential and Frequent Virus Latency in Blood-Derived T-Cell Clones Infected With Human T-Cell Leukemia Virus Type I

Blood ◽

10.1182/blood.v89.9.3303 ◽

1997 ◽

Vol 89 (9) ◽

pp. 3303-3314 ◽

Cited By ~ 45

Author(s):

J.H. Richardson ◽

P. Höllsberg ◽

A. Windhagen ◽

L.A. Child ◽

D.A. Hafler ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Leukemia Virus ◽

Type I ◽

Cell Leukemia Virus Type ◽

T Cell Clones ◽

Human T Cell ◽

Cell Clones ◽

T Cell Leukemia Virus ◽

Spontaneous Proliferation

Abstract Human T-cell leukemia virus type I (HTLV-I)-infected T cells expanded in vitro by single-cell cloning provide a unique system for investigating virus-cell interactions in nonimmortalized T cells. By analysis of clones generated randomly from the blood of virus carriers, we confirm that CD4 T cells are the major reservoir of HTLV-I in vivo and show that most infected cells contain a single integrated provirus. Contrary to the situation in HTLV-I immortalized cell lines, the HTLV-I provirus was found to be transcriptionally silent in a high proportion of randomly generated T-cell clones and could not be reactivated by mitogenic stimulation. The spontaneous proliferation previously documented in HTLV-I–infected T-cell clones was not observed in silently infected cells, and therefore correlates directly with the expression of tax and other viral genes. The only cytokine mRNA found to be significantly elevated in the virus-producing clones was interleukin-6; however, receptor-blocking experiments argue against a role for IL-6 in the virus-induced cell proliferation. We observed a striking variation in the ability of individual HTLV-I–producing clones to immortalize fresh peripheral blood lymphocytes. This ability did not correlate with the levels of viral mRNA expression, gag p24 production, spontaneous proliferation, or tax-transactivation, possibly suggesting a role for host cell factors as determinants of viral infectivity or immortalization. Studies to elucidate the basis of this phenotypic heterogeneity should enhance our understanding of viral spread and pathogenesis.

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Diverse usage of human T-cell receptor gene segments in HLA-DR1 allospecific T-cell clones

Human Immunology ◽

10.1016/0198-8859(96)00060-2 ◽

1996 ◽

Vol 49 (2) ◽

pp. 122-129 ◽

Cited By ~ 7

Author(s):

Masao Ota ◽

Mary Jane Geiger ◽

Sandra Rosen-Bronson ◽

Carolyn Katovich Hurley ◽

David D. Eckels

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Receptor Gene ◽

Cell Receptor ◽

T Cell Clones ◽

Human T Cell ◽

Cell Clones ◽

T Cell Receptor Gene ◽

Cell Receptor Gene

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Human T-cell receptor TCRAV, TCRBV, and TCRAJ sequences newly found in T-cell clones reactive with allogeneic HLA class II antigens

Immunogenetics ◽

10.1007/bf00216395 ◽

1993 ◽

Vol 38 (1) ◽

pp. 67-70 ◽

Cited By ~ 8

Author(s):

Fumiya Obata ◽

Misao Tsunoda ◽

Takehisa Kaneko ◽

Koichi Ito ◽

Ichiro Ito ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Class Ii ◽

Hla Class Ii ◽

T Cell Clones ◽

Human T Cell ◽

Cell Clones ◽

Class Ii Antigens

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Human T-Cell Leukemia Virus Type 1 (HTLV-1) Infection of Mice: Proliferation of Cell Clones with Integrated HTLV-1 Provirus in Lymphoid Organs

Journal of Virology ◽

10.1128/jvi.75.9.4420-4423.2001 ◽

2001 ◽

Vol 75 (9) ◽

pp. 4420-4423 ◽

Cited By ~ 10

Author(s):

Masakazu Tanaka ◽

Binlian Sun ◽

Jianhua Fang ◽

Takayuki Nitta ◽

Toshinori Yoshida ◽

...

Keyword(s):

T Cell ◽

Leukemia Virus ◽

T Cell Leukemia ◽

Cell Leukemia ◽

Cell Leukemia Virus Type ◽

Human T Cell ◽

Cell Clones ◽

T Cell Leukemia Virus ◽

Flanking Sequences

ABSTRACT Human T-cell leukemia virus type 1 (HTLV-1) is suggested to cause adult T-cell leukemia after 40 to 50 years of latency in a small percentage of carriers. However, little is known about the pathophysiology of the latent period and the reservoir organs where polyclonal proliferation of cells harboring integrated provirus occurs. The availability of animal models would be useful to analyze the latent period of HTLV-1 infection. At 18 months after HTLV-1 infection of C3H/HeJ mice inoculated with the MT-2 cell line, which is an HTLV-1-producing human T-cell line, HTLV-1 provirus was detected in spleen DNA from eight of nine mice. No more than around 100 proviruses were found per 105 spleen cells. Cellular sequences flanking the 3′ long terminal repeat (LTR) and the clonalities of the cells which harbor integrated HTLV-1 provirus were analyzed by linker-mediated PCR. The results showed that the flanking sequences are of mouse genome origin and that polyclonal proliferation of the spleen cells harboring integrated HTLV-1 provirus had occurred in three mice. A sequence flanking the 5′ LTR was isolated from one of the mice and revealed the presence of a 6-nucleotide duplication of cellular sequences, consistent with typical retroviral integration. Moreover, PCR was performed on DNA from infected tissues, with LTR primers and primers derived from seven novel flanking sequences of the three mice. Data revealed that the expected PCR products were found from lymphatic tissues of the same mouse, suggesting that the lymphatic tissues were the reservoir organs for the infected and proliferating cell clones. The mouse model described here should be useful for analysis of the carrier state of HTLV-1 infection in humans.

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