Peptide-Specific, TCR-α–Driven, Coreceptor-Independent Negative Selection in TCR α-Chain Transgenic Mice

Furmanski, Anna L.; Bartók, I; Chai, Jian-Guo; Singh, Yogesh; Ferreira, Cristina; Scott, Diane; Holland, Stephen J.; Bourdeaux, Christophe; Crompton, Tessa; Dyson, Julian

doi:10.4049/jimmunol.0902291

Cited by 3 publications

(3 citation statements)

References 45 publications

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“…First, to introduce wide-ranging structural variation into the CDRs in vivo, our TCR-β chain retrogenic approach (14,15) was modified to direct V(D)J recombination into the germ-line CDRs. This was achieved by appropriate positioning of recombination signal sequences (RSSs) and an intervening spacer into the center of the targeted germ-line CDR.…”

Section: Resultsmentioning

confidence: 99%

The T-cell receptor is not hardwired to engage MHC ligands

Holland

Bartók

Attaf

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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The bias of αβ T cells for MHC ligands has been proposed to be intrinsic to the T-cell receptor (TCR). Equally, the CD4 and CD8 coreceptors contribute to ligand restriction by colocalizing Lck with the TCR when MHC ligands are engaged. To determine the importance of intrinsic ligand bias, the germ-line TCR complementarity determining regions were extensively diversified in vivo. We show that engagement with MHC ligands during thymocyte selection and peripheral T-cell activation imposes remarkably little constraint over TCR structure. Such versatility is more consistent with an opportunist, rather than a predetermined, mode of interface formation. This hypothesis was experimentally confirmed by expressing a hybrid TCR containing TCR-γ chain germ-line complementarity determining regions, which engaged efficiently with MHC ligands. MHC restriction | TCRT cells expressing an αβ T-cell receptor (TCR) are MHCrestricted, recognizing self-and foreign peptide epitopes presented by MHC class I and II molecules during thymic development and peripheral activation, respectively. Two mechanisms are proposed to underlie this ligand bias. First, the CD8 and CD4 coreceptors have dual specificity for extracellular MHC and the intracellular proximal kinase Lck. Consequently, when MHC class I or II ligands are engaged, Lck is colocalized with the TCR/CD3 complex initiating signal transduction. The importance of this mechanism in disadvantaging non-MHC ligands is highlighted by the recovery of T-cell selection, where MHC and the coreceptors are both absent in comparison to the absence of MHC alone (1). In this setting, non-MHC ligands drive thymic positive selection and are recognized by peripheral T cells (2). The ability of αβ TCRs to recognize non-MHC ligands does not rule out an intrinsic bias of the TCR for MHC ligands. Indeed, evidence for such a hardwired bias is suggested by pairwise interactions between TCR-β germ-line complementarity determining regions 1 and 2 (CDR1/2) and the MHC α-helices observed in several structures (3-7). Recently, such recurrent interactions have been shown to be dependent on the partner TCR-α chain, which can impose distinct modes of TCR-β engagement, suggesting they may not drive MHC specificity (8). Although the relatively limited set of TCR/MHCpeptide structures reveals a semiconserved docking geometry, the angle of TCR engagement varies by more than 60°and the generally central docking position can shift toward the peptide aminoor carboxy-terminus (9). Likewise, conserved features of the MHC α-helices, including exposure of the polypeptide backbone and surface depressions, have been suggested to provide energetically favorable sites for CDR engagement (5). However, a crucial role for specific MHC residues in TCR docking has not emerged (10). The role of germ-line TCR structure in the bias to MHC ligands has thus been perplexing, especially given the structural variability of both components. To investigate this, we have applied a unique mutagenesis approach based on redirecting V(D)J recombina...

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Section: Resultsmentioning

confidence: 99%

The T-cell receptor is not hardwired to engage MHC ligands

Holland

Bartók

Attaf

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Down-modulation of CD4 and CD8 co-receptors has been observed in other transgene models of αβ T cell tolerance and has been proposed as a potential censorship mechanism for self-reactive cells (31, 32). Alternatively, peripheral DN T cells could be artifacts of premature expression of the transgenic αβTCR at the DN stage of thymocyte development as mentioned above (33). Some authors have argued that prematureαβTCR expression may preempt co-receptor expression and MHC-selection in the thymus and allow peripheral migration of αβ analogues of γδ T cells (34, 35).…”

Section: Discussionmentioning

confidence: 99%

Self-Tolerance Checkpoints in CD4 T Cells Specific for a Peptide Derived from the B Cell Antigen Receptor

Detanico

Heiser

Aviszus

et al. 2011

The Journal of Immunology

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Linked recognition of antigen by B and T lymphocytes is ensured in part by a state of tolerance acquired by CD4 T cells to germline-encoded sequences within the B cell antigen receptor (BCR). We sought to determine how such tolerance is attained when a peptide from the BCR variable (V) region is expressed by small numbers of B cells as it is in the physiological state. Mixed bone marrow (BM) chimeras were generated using donor BM from mice with B cells that expressed a transgene (Tg)-encoded kappa light chain and BM from T cell receptor (TCR) Tg mice in which the CD4 T cells (CA30) were specific for a Vκ peptide encoded by the kappa transgene (κTg). In chimeras where few B cells express the κTg, many CA30 cells were deleted in the thymus. However, a substantial fraction survived to the CD4 single-positive stage. Among single-positive CA30 thymocytes, few reached maturity and migrated to the periphery. Maturation was strongly associated with, and likely promoted by, expression of an endogenous TCR alpha chain. CD4+ CA30 cells that reached peripheral lymphoid tissues were antigen-experienced and anergic, and some developed into regulatory cells. These findings reveal several checkpoints and mechanisms that enforce a state of self-tolerance in developing T cells specific for BCR V region sequences, thus ensuring that T cell help to B cells occurs through linked recognition of foreign antigen.

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“…19,20,40,41,43 The use of TCR retrogenic mice has facilitated the analysis of specific T-cell populations in vivo in a variety of areas, particularly autoimmunity (Table 2). 19,20,[40][41][42][43][46][47][48][49][50][51][52][53][54][55][56][57] Initially, we performed proof-of-principle experiments to demonstrate that the development and function of antigen-specific T cells, such as the ovalbumin-specific TCR OT-I and OT-II, were comparable between TCR retrogenic and transgenic mice. 19,20 Retroviral expression of both TCRs resulted in the expected skewing to either the CD8 + or CD4 + lineage, and peripheral T-cell accumulation as early as 5 weeks after bone marrow transfer.…”

Section: Multicistronic 'Self-cleaving' 2a Peptide-based Retroviral Vmentioning

confidence: 99%

T‐cell receptor retrogenic mice: a rapid, flexible alternative to T‐cell receptor transgenic mice

Bettini

Vignali

2012

Immunology

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The T-cell receptor (TCR) is unique in its complexity. It determines not only positive (life) and negative (death) selection in the thymus, but also mediates proliferation, anergy, differentiation, cytotoxicity and cytokine production in the periphery. Through its association with six CD3 signalling chains (εγ, δε and ζζ), the TCR is capable of recognizing an extensive variety of antigenic peptides, from both pathogens and self-antigens, and translating these interactions into multiple signalling pathways that mediate diverse T-cell developmental and functional responses. The analysis of TCR biology has been revolutionized by the development of TCR transgenic mice, which express a single clonotypic T-cell population, with diverse specificities and genetic backgrounds. However, they are time consuming to generate and characterize, limiting the analysis of large numbers of TCR over a short period of time in multiple genetic backgrounds. The recent development of TCR retrogenic technology resolves these limitations and could in time have a similarly important impact on our understanding of T-cell development and function. In this review, we will discuss the advantages and limitations of retrogenic technology compared with the generation and use of TCR transgenic mice for studying all aspects of T-cell biology.

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Peptide-Specific, TCR-α–Driven, Coreceptor-Independent Negative Selection in TCR α-Chain Transgenic Mice

Cited by 3 publications

References 45 publications

The T-cell receptor is not hardwired to engage MHC ligands

The T-cell receptor is not hardwired to engage MHC ligands

Self-Tolerance Checkpoints in CD4 T Cells Specific for a Peptide Derived from the B Cell Antigen Receptor

T‐cell receptor retrogenic mice: a rapid, flexible alternative to T‐cell receptor transgenic mice

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