The CD3ε Proline-Rich Sequence, and Its Interaction with Nck, Is Not Required for T Cell Development and Function

Szymczak, Andrea L; Workman, Creg J.; Gil, Diana; Dilioglou, Smaroula; Vignali, Kate M.; Palmer, Ed

doi:10.4049/jimmunol.175.1.270

Cited by 60 publications

(72 citation statements)

References 46 publications

(37 reference statements)

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“…No gross abnormalities were observed in the distribution of thymic populations, in accordance with our observation that the conformational change is not elicited during positive selection. Unfortunately this system does not offer information regarding the role of the CD3 proline-rich sequence in negative selection because it was not studied in transgenic TCR models (27).…”

Section: Discussionmentioning

confidence: 99%

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A conformational change senses the strength of T cell receptor–ligand interaction during thymic selection

Risueño

Santen

Alarcón³

2006

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

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T cell antigen receptor (TCR) triggering determines the fate of immature thymocytes. The affinity of the TCR for its endogenous peptide͞MHC ligands serves as a signal for positive or negative selection through mechanisms that are still little understood. We have used a conformation-specific antibody to demonstrate that recognition of TCR ligands that lead to negative selection induces a conformational change in the TCR in situ. In contrast, this conformational change is elicited in only a small percentage of immature thymocytes during positive selection. Using a TUNEL assay, we demonstrate that the conformational change in the TCR is strongly linked to activation of programmed cell death in conditions leading to negative selection. Furthermore, the few conformational change-positive thymocytes detected in conditions that preferably lead to positive selection are also TUNELpositive. Thus, the conformational change in the TCR may underlie the discrimination of ligands leading to positive and negative selection.signal transduction ͉ thymic differentiation ͉ negative selection T he T cell antigen receptor (TCR) complex is composed in most T cells of the ligand-binding TCR␣ and TCR␤ subunits and the signal-transducing CD3␥, CD3␦, CD3 , and CD3 (CD247) (1, 2). The differentiation of T cells is controlled by the transduction of pre-TCR and TCR signals. Differentiation of immature CD4 ϩ CD8 ϩ double-positive (DP) to mature CD4 ϩ or CD8 ϩ single-positive thymocytes is initiated when the TCR-␣␤ heterodimer engages the peptide͞MHC (pMHC) (2). A unique feature of the TCR is its ability to scan structurally similar pMHC ligands and transmit distinct biochemical signals depending on the strength of the ligand recognized (3, 4). In developing thymocytes, weak TCR ligands induce positive selection, and stronger ligands induce negative selection (5). However, mature T cells are tolerant to weak TCR ligands in the periphery, and they respond to strong ligands by activating a program of cell proliferation and cell differentiation. The differential responsiveness of immature and mature T cells to weak ligands constitutes the cornerstone of self-tolerance that is acquired in the thymus.Many studies have focused on how the TCR␣␤ heterodimer communicates ligand engagement to the CD3 subunits, how the CD3 subunits translate this information into a specific cellular response, and how the TCR complex discriminates between strong and weak ligands. The engagement of the TCR with a weak ligand in mature T cells results in a distinct pattern of intracellular signaling compared with that induced by a full-agonist peptide. For example, stimulation with a weak ligand induces only partial phosphorylation of the CD3 and CD3 chains and the recruitment, but not the phosphorylation, of the -associated protein of 70 kDa, ZAP70 (6, 7). Hence, weak ligands activate only a subset of the downstream signaling pathways that are activated by an agonist peptide (8). Weak ligands bind to the TCR with a lower affinity and a higher dissociation rate than full a...

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Section: Discussionmentioning

confidence: 99%

“…The importance of the CD3 proline-rich sequence in thymic development has recently been brought into question (27). Bone marrow chimeras in CD3 -deficient mice have been used to express a CD3 mutated in the proline-rich sequence before studying T cell development.…”

Section: Discussionmentioning

confidence: 99%

A conformational change senses the strength of T cell receptor–ligand interaction during thymic selection

Risueño

Santen

Alarcón³

2006

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

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“…Systems using 293T transfected cells such as Phoenix (Phoenix-Ampho, American Type Culture Collection (ATCC) SD3443) and Eco (ATCC: SD 3444), which can be directly transfected, are most commonly used. However, our lab has successfully used a 3T3-based cell line, GP+E86 (ATCC: CRL-9642) 7,11,14 . The two-step process we describe here first generates retrovirus by the transient transfection of 293T cells with three plasmids, to minimize the possibility of generating replicationcompetent retrovirus (RCR) 15,16 .…”

Section: Generation Of Retroviral Producer Cell Linesmentioning

confidence: 99%

Generation of T-cell receptor retrogenic mice

et al. 2006

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“…Bone marrow reconstitution with CD3ε PRS mutants in CD3ε-deficient mice, Nck overexpression in primary T cells, Nck knockout mice, and PRS knockin (KI) mice have all been used to study the role of the PRS and Nck in T cell development and mature T cell activation (14)(15)(16)(17)(18)(19)(20). Some experiments have suggested that the PRS is important for thymic maturation, but not for mature T cell activation in vitro (15), although experiments with bone marrow chimeras indicate that the PRS is important to activate mature T cells by weak, but not by strong, agonists in vitro (19).…”

mentioning

confidence: 99%

Relevance of Nck–CD3ε Interaction for T Cell Activation In Vivo

Borroto

Arellano

Blanco

et al. 2014

The Journal of Immunology

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On TCR ligation, the adaptor Nck is recruited through its src homology 3.1 domain to a proline-rich sequence (PRS) in CD3ε. We have studied the relevance of this interaction for T cell activation in vitro and in vivo by targeting the interaction sites in both partners. The first approach consisted of studying a knockin (KI) mouse line (KI-PRS) bearing a conservative mutation in the PRS that makes the TCR incompetent to recruit Nck. This deficiency prevents T cell activation by Ag in vitro and inhibited very early TCR signaling events including the tyrosine phosphorylation of CD3ζ. Most important, KI-PRS mice are partly protected against the development of neurological symptoms in an experimental autoimmune encephalitis model, and show a deficient antitumoral response after vaccination. The second approach consisted of using a high-affinity peptide that specifically binds the src homology 3.1 domain and prevents the interaction of Nck with CD3ε. This peptide inhibits T cell proliferation in vitro and in vivo. These data suggest that Nck recruitment to the TCR is fundamental to mount an efficient T cell response in vivo, and that the Nck–CD3ε interaction may represent a target for pharmacological modulation of the immune response.

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The CD3ε Proline-Rich Sequence, and Its Interaction with Nck, Is Not Required for T Cell Development and Function

Cited by 60 publications

References 46 publications

A conformational change senses the strength of T cell receptor–ligand interaction during thymic selection

A conformational change senses the strength of T cell receptor–ligand interaction during thymic selection

Generation of T-cell receptor retrogenic mice

Relevance of Nck–CD3ε Interaction for T Cell Activation In Vivo

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