Class I MHC molecules on hematopoietic cells can support intrathymic positive selection of T cell receptor transgenic T cells

We have developed a unique in vivo system to determine the relationship between endogenous altered peptide ligands and the development of major histocompatibility complex class II– restricted T cells. Our studies use the 3.L2 T cell receptor (TCR) transgenic mouse, in which T cells are specific for Hb(64–76)/I-Ek and positively selected on I-Ek plus self-peptides. To this endogenous peptide repertoire, we have individually added one of six well-characterized 3.L2 ligands. This transgenic approach expands rather than constrains the repertoire of self-peptides. We find that a broad range of ligands produce negative selection of thymocytes in vivo. When compared with the in vitro TCR–ligand binding kinetics, we find that these negatively selecting ligands all have a half-life of 2 s or greater. Additionally, one of two ligands examined with no detectable binding to the 3.L2 TCR and no activity on mature 3.L2 T cells (Q72) enhances the positive selection of transgenic thymocytes in vivo. Together, these data establish a kinetic threshold between negative and positive selection based on the longevity of TCR–ligand complexes.

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Phenotypic and functional analysis of positive selection in the gamma/delta T cell lineage.

Journal of Experimental Medicine ◽

10.1084/jem.177.4.1061 ◽

1993 ◽

Vol 177 (4) ◽

pp. 1061-1070 ◽

Cited By ~ 17

Author(s):

F B Wells ◽

Y Tatsumi ◽

J A Bluestone ◽

S M Hedrick ◽

J P Allison ◽

...

Keyword(s):

Signal Transduction ◽

T Cells ◽

T Cell ◽

Positive Selection ◽

Class I ◽

Gamma Delta ◽

Class I Mhc ◽

General Role ◽

Alpha Beta ◽

Selection Of

Recent evidence suggests that T cells expressing gamma/delta antigen receptors (T cell receptor [TCR]) are subject to positive selection during development. We have shown that T cells expressing a class I major histocompatibility complex (MHC)-specific gamma/delta TCR transgene (tg) are not positively selected in class I MHC-deficient, beta 2-microglobulin (beta 2m) gene knockout mice (tg+ beta 2m-). In this report, we examine phenotypic and functional parameters of gamma/delta positive selection in this transgenic model system. TCR-gamma/delta tg+ thymocytes of mature surface phenotype (heat stable antigen-, CD5hi) were found in beta 2m+ but not in beta 2m- mice. Moreover, subsets of tg+ thymocytes with the phenotype of activated T cells (interleukin [IL]2R+, CD44hi, or Mel-14lo) were also present only in the beta 2m+ mice. Cyclosporine A, which blocks positive selection of TCR-alpha/beta T cells, also inhibited gamma/delta tg+ T cell development. These results support the idea that positive selection of TCR-gamma/delta requires active TCR-mediated signal transduction. Whereas tg+ beta 2m+ thymocytes produced IL-2 and proliferated when stimulated by alloantigen, TCR engagement of tg+ beta 2m- thymocytes by antigen induced IL-2R expression but was uncoupled from the signal transduction pathway leading to IL-2 production and autocrine proliferation. Overall, these results demonstrate significant parallels between gamma/delta and alpha/beta lineage development, and suggest a general role for TCR signaling in thymic maturation.

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Distinct phases in the positive selection of CD8+ T cells distinguished by intrathymic migration and T-cell receptor signaling patterns

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1408482111 ◽

2014 ◽

Vol 111 (25) ◽

pp. E2550-E2558 ◽

Cited By ~ 30

Author(s):

J. O. Ross ◽

H. J. Melichar ◽

B. B. Au-Yeung ◽

P. Herzmark ◽

A. Weiss ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Positive Selection ◽

T Cell Receptor ◽

Cd8 T Cells ◽

Cell Receptor ◽

Receptor Signaling ◽

T Cell Receptor Signaling ◽

Cell Receptor Signaling ◽

Selection Of

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Early Deletion and Late Positive Selection of T-Cells Expressing a Male-Specific Receptor in T-Cell Receptor Transgenic Mice

Developmental Immunology ◽

10.1155/1990/18208 ◽

1990 ◽

Vol 1 (1) ◽

pp. 1-10 ◽

Cited By ~ 52

Author(s):

Hung Sia Teh ◽

Hiroyuki Kishi ◽

Bernadette Scott ◽

Peter Borgulya ◽

Harald Von Boehmer ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Transgenic Mice ◽

Positive Selection ◽

T Cell Receptor ◽

Interleukin 2 ◽

Cell Receptor ◽

Early Expression ◽

Single Positive ◽

Selection Of

The ontogeny of T cells in T-cell receptor (TCR) transgenic mice, which express a transgenicαβheterodimer, specific for the male (H-Y) antigen in association with H-2Db, was determined. The transgenicαchain was expressed on about 10% of the fetal thymocytes on day 14 of gestation. About 50% of day-15 fetal thymocytes expressed bothαandβtranschains and virtually all fetal thymocytes expressed the transgenicαβheterodimer by day 17. The early expression of the transgenic TCR on CD4-8-thymocytes prevented the development ofγδcells, and led to accelerated growth of thymocytes and an earlier expression of CD4 and CD8 molecules. Up to day 17, no significant differences in T-cell development could be detected between female and male thymuses. By day 18 of gestation, the male transgenic thymus contained more CD4-8-thymocytes than the female transgenic thymus. The preponderance of CD4-8-thymocytes in the male transgenic thymus increased until birth and was a consequence of the deletion of the CD4+8+thymocytes and their CD4-8+precursors. By the time of birth, the male transgenic thymus contained half the number of cells as the female transgenic thymus. The deletion of autospecific precursor cells in the male transgenic mouse began only at day 18 of gestation, despite the fact that the ligand could already be detected by day 16.The preferential accumulation of CD4-8+T cells, which expressed a high density of the transgenic TCR, occurred only after birth and was .obvious in 6-week-old female thymus. These data support the hypothesis that the positive selection of T cells expressing this transgenic heterodimer may involve two steps, i.e., the commitment of CD4+8+thymocytes to the CD4-8+lineage following the interaction of the transgenic TCR with restricting major histocompatibility molecules, followed by a slow conversion of CD4+8+thymocytes into CD4-8+T cells.In normal mice, the precursors of CD+4+8 and single positive thymocytes have the CD4-8-CD3-J11d+(or M1/69+) phenotype. Because of the early expression of the transgenicαβheterodimer, this population was not detected in adult transgenic mice. All CD4-8-M1/ 69+cells expressed the transgenic receptor associated with CD3 and could be readily grown in media containing T-cell lectins and interleukin 2.

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Positive selection determines T cell receptor V beta 14 gene usage by CD8+ T cells.

Journal of Experimental Medicine ◽

10.1084/jem.170.1.135 ◽

1989 ◽

Vol 170 (1) ◽

pp. 135-143 ◽

Cited By ~ 80

Author(s):

N S Liao ◽

J Maltzman ◽

D H Raulet

Keyword(s):

T Cells ◽

T Cell ◽

Positive Selection ◽

T Cell Receptor ◽

Cd8 T Cells ◽

Cd4 T Cells ◽

Cell Receptor ◽

Radiation Chimeras ◽

Gene Usage ◽

Selection Of

We report here a mAb, 14-2, reactive with TCRs that include V beta 14. The frequency of V beta 14+ T cells varies with CD4 and CD8 subset and is controlled by the H-2 genes. Thus CD8+ T cells from H-2b mice include approximately 2.3% V beta 14+ T cells while CD8+ T cells from mice expressing K kappa include greater than 8% V beta 14+ T cells. In all strains examined, 7-8% of CD4+ T cells express V beta 14. The frequent usage of V beta 14 in CD8+ T cells of K kappa-expressing mice is a result of preferential positive selection of V beta 14+ CD8+ T cells as demonstrated by analysis of radiation chimeras. These studies demonstrate that H-2-dependent positive selection occurs in unmanipulated mice. Furthermore, the results imply that positive selection, and possibly H-2 restriction, can be strongly influenced by a V beta domain, with some independence from the beta-junctional sequence and alpha chain.

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Clone-specific T cell receptor antagonists of major histocompatibility complex class I-restricted cytotoxic T cells.

Journal of Experimental Medicine ◽

10.1084/jem.177.6.1541 ◽

1993 ◽

Vol 177 (6) ◽

pp. 1541-1550 ◽

Cited By ~ 199

Author(s):

S C Jameson ◽

F R Carbone ◽

M J Bevan

Keyword(s):

T Cells ◽

Major Histocompatibility Complex ◽

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Class I ◽

Major Histocompatibility ◽

Class I Mhc ◽

Histocompatibility Complex ◽

T Cell Clone

A previous report showed that the proliferative response of helper T cells to class II major histocompatibility complex (MHC)-restricted antigens can be inhibited by analogues of the antigen, which act as T cell receptor (TCR) antagonists. Here we define and analyze peptide variants that antagonize various functions of class I MHC-restricted cytotoxic T lymphocyte (CTL) clones. Of 64 variants at individual TCR contact sites of the Kb-restricted octamer peptide ovalbumin257-264 (OVAp), a very high proportion (40%) antagonized lysis by three OVAp-specific CTL clones. This effect was highly clone specific, since many antagonists for one T cell clone have differential effects on another. We show that this inhibition of CTL function is not a result of T cell-T cell interaction, precluding veto-like phenomena as a mechanism for antagonism. Moreover, we present evidence for direct interaction between the TCR and antagonist-MHC complexes. In further analysis of the T cell response, we found that serine esterase release and cytokine production are susceptible to TCR antagonism similarly to lysis. Ca2+ flux, an early event in signaling, is also inhibited by antagonists but may be more resistant to the antagonist effect than downstream responses.

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Increased frequency of T cell receptor V alpha 12.1 expression on CD8+ T cells: evidence that V alpha participates in shaping the peripheral T cell repertoire.

Journal of Experimental Medicine ◽

10.1084/jem.174.3.639 ◽

1991 ◽

Vol 174 (3) ◽

pp. 639-648 ◽

Cited By ~ 52

Author(s):

H DerSimonian ◽

H Band ◽

M B Brenner

Keyword(s):

T Cells ◽

T Cell ◽

Gene Product ◽

T Cell Receptor ◽

Cd8 T Cells ◽

Peripheral Blood ◽

Cell Receptor ◽

Class I ◽

Class I Mhc ◽

Alpha Gene

The T cell receptor repertoire has a potential for vast diversity. However, this diversity is limited by the fact that the majority of thymocytes die as the repertoire is shaped by positive and negative selection events during development. Such thymic selection affecting TCR V beta gene segment usage has been demonstrated in the mouse. However, similar data has not been forthcoming in man, and little is known about the role of the TCR alpha chain in antigen/major histocompatibility complex (MHC) recognition in any species. Here, we used a monoclonal antibody recognizing the TCR V alpha 12.1 gene product to assess the expression of this gene in the peripheral blood of man. In most individuals tested, the percentage of cells expressing V alpha 12.1 was significantly higher in CD8+ T cells than in CD4+ T cells. That the V alpha gene product itself was responsible for this increased expression in CD8+ T cells was underscored by the lack of substantial skewing of V beta usage in the V alpha 12.1-bearing T cells. Moreover, the skewed expression of V alpha 12.1 was already present at birth, indicating that it was likely to be due to a developmental process rather than the result of exposure to environmental antigens. Based on the established role for CD8 in binding to class I MHC molecules, we suggest that increased expression of V alpha 12.1 on CD8+ T cells points to a role for TCR's using V alpha 12.1 in class I MHC/Ag recognition. These results indicate that V alpha gene usage in the peripheral blood of man is not random, and they support a role for V alpha as a participant in the self-MHC recognition process that shapes the TCR repertoire.

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Increased p300 Expression Inhibits Glucocorticoid Receptor-T-Cell Receptor Antagonism but Does Not Affect Thymocyte Positive Selection

Molecular and Cellular Biology ◽

10.1128/mcb.22.13.4556-4566.2002 ◽

2002 ◽

Vol 22 (13) ◽

pp. 4556-4566 ◽

Cited By ~ 7

Author(s):

Cheng-Tai Yu ◽

Ming-Hsien Lin Feng ◽

Hsiu-ming Shih ◽

Ming-Zong Lai

Keyword(s):

T Cells ◽

T Cell ◽

Transgenic Mice ◽

Glucocorticoid Receptor ◽

Positive Selection ◽

T Cell Receptor ◽

Critical Role ◽

Interleukin 2 ◽

Cell Receptor ◽

Selection Of

ABSTRACT Positive selection of T cells is postulated to be dependent on the counterinteraction between glucocorticoid receptor (GR)- and T-cell-receptor (TCR)-induced death signals. In this study we used T-cell-specific expression of p300 to investigate whether GR-TCR cross talk between thymocytes was affected. Activation of the p300-transgenic T cells led to enhanced thymocyte proliferation and increased interleukin 2 production. Thymocyte death, induced by TCR engagement, was no longer prevented by dexamethasone in p300-transgenic mice, indicating an absence of GR-TCR cross-inhibition. This was accompanied by a 50% reduction in the number of thymocytes in p300-transgenic mice. However, the CD4/CD8 profile of thymocytes remained unchanged in p300-transgenic mice. There was no effect on positive selection of the bulk thymocytes or thymocytes with transgenic TCR in p300-transgenic mice. In addition, there was no apparent TCR repertoire “hole” in the selected antigens examined. Our results illustrate a critical role of CBP/p300 in thymic GR-TCR counterinteraction yet do not support the involvement of GR-TCR antagonism in thymocyte positive selection.

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