Biophysical mechanism of T-cell receptor triggering in a reconstituted system

James, John R.; Vale, Ronald D.

doi:10.1038/nature11220

Cited by 305 publications

(353 citation statements)

References 38 publications

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“…Notably, in addition to the conformational dynamics model shown here, other models such as kinetic segregation and kinase regulation have been shown to be crucial in TCR activation [7,8,76,77]. Our finding does not contradict other models but rather support the notion that different factors act together to generate a multi-layered regulation of TCR activation.…”

Section: Discussionsupporting

confidence: 75%

“…The kinase-phosphatase equilibrium should play a key role in Step 2 in regulating ITAM phosphorylation. Kinetic segregation of phosphatases and the local enrichment of active Lck are both important to initiate TCR phosphorylation [7,8,76,77]. Altogether, the initial TCR triggering is regulated by a complex mechanism and modulation of either lipid binding-dissociation equilibrium or kinase-phosphatase equilibrium can affect the final outcome of TCR activation.…”

Section: Discussionmentioning

confidence: 99%

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Lipid-dependent conformational dynamics underlie the functional versatility of T-cell receptor

Guo

Yan

et al. 2017

Cell Res

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T-cell receptor-CD3 complex (TCR) is a versatile signaling machine that can initiate antigen-specific immune responses based on various biochemical changes of CD3 cytoplasmic domains, but the underlying structural basis remains elusive. Here we developed biophysical approaches to study the conformational dynamics of CD3ε cytoplasmic domain (CD3ε CD ). At the single-molecule level, we found that CD3ε CD could have multiple conformational states with different openness of three functional motifs, i.e., ITAM, BRS and PRS. These conformations were generated because different regions of CD3ε CD had heterogeneous lipid-binding properties and therefore had heterogeneous dynamics. Live-cell imaging experiments demonstrated that different antigen stimulations could stabilize CD3ε CD at different conformations. Lipid-dependent conformational dynamics thus provide structural basis for the versatile signaling property of TCR.

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Section: Discussionsupporting

confidence: 75%

Section: Discussionmentioning

confidence: 99%

Lipid-dependent conformational dynamics underlie the functional versatility of T-cell receptor

Guo

Yan

et al. 2017

Cell Res

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“…The relationship between the immune synapse gap and the efficiency of T-cell activation and cytoIysis is well documented. 21 Consistent with these observations, it has been reported that the size of the extracellular domain of antigens targeted by BiTEs influences the efficacy of the cell killing, such that molecules with large extracellular domains do not lead to efficient redirected cell lysis. 22 In a similar way, it could be hypothesized that the size of the bispecific molecules could affect the potency of the cell killing, with larger molecules being less efficient than smaller molecules.…”

Section: Discussionsupporting

confidence: 52%

bisFabs: Tools for rapidly screening hybridoma IgGs for their activities as bispecific antibodies

Patke¹,

Li²,

Wang³

et al. 2017

mAbs

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Bispecific antibodies are a growing class of therapeutic molecules. Many of the current bispecific formats require DNA engineering to convert the parental monoclonal antibodies into the final bispecific molecules. We describe here a method to generate bispecific molecules from hybridoma IgGs in 3-4 d using chemical conjugation of antigen-binding fragments (Fabs) (bisFabs). Proteolytic digestion conditions for each IgG isotype were analyzed to optimize the yield and quality of the final conjugates. The resulting bisFabs showed no significant amounts of homodimers or aggregates. The predictive value of murine bisFabs was tested by comparing the T-cell redirected cytotoxic activity of a panel of antibodies in either the bisFab or full-length IgG formats. A variety of antigens with different structures and expression levels was used to extend the comparison to a wide range of binding geometries and antigen densities. The activity observed for different murine bisFabs correlated with those observed for the fulllength IgG format across multiple different antigen targets, supporting the use of bisFabs as a screening tool. Our method may also be used for the screening of bispecific antibodies with other mechanisms of action, allowing for a more rapid selection of lead therapeutic candidates.

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“…These data suggest that the hRSV N may inhibit IS assembly via mechanisms that take advantage of the low affinity of the physiological TCR−pMHC interaction. For example, N may (i) impair bending of the T-cell membrane necessary to form a close contact (44), or (ii) impair physiological microclustering of newly formed monovalent pMHC−TCR pairs.…”

Section: Resultsmentioning

confidence: 99%

Surface expression of the hRSV nucleoprotein impairs immunological synapse formation with T cells

Céspedes

Bueno

Ramírez

et al. 2014

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

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Human respiratory syncytial virus (hRSV) is the leading cause of bronchiolitis and pneumonia in young children worldwide. The recurrent hRSV outbreaks and reinfections are the cause of a significant public health burden and associate with an inefficient antiviral immunity, even after disease resolution. Although several mouse-and human cell-based studies have shown that hRSV infection prevents naïve T-cell activation by antigen-presenting cells, the mechanism underlying such inhibition remains unknown. Here, we show that the hRSV nucleoprotein (N) could be at least partially responsible for inhibiting T-cell activation during infection by this virus. Early after infection, the N protein was expressed on the surface of epithelial and dendritic cells, after interacting with trans-Golgi and lysosomal compartments. Further, experiments on supported lipid bilayers loaded with peptide-MHC (pMHC) complexes showed that surface-anchored N protein prevented immunological synapse assembly by naive CD4 + T cells and, to a lesser extent, by antigen-experienced T-cell blasts. Synapse assembly inhibition was in part due to reduced T-cell receptor (TCR) signaling and pMHC clustering at the T-cell− bilayer interface, suggesting that N protein interferes with pMHC− TCR interactions. Moreover, N protein colocalized with the TCR independently of pMHC, consistent with a possible interaction with TCR complex components. Based on these data, we conclude that hRSV N protein expression at the surface of infected cells inhibits T-cell activation. Our study defines this protein as a major virulence factor that contributes to impairing acquired immunity and enhances susceptibility to reinfection by hRSV.

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Biophysical mechanism of T-cell receptor triggering in a reconstituted system

Cited by 305 publications

References 38 publications

Lipid-dependent conformational dynamics underlie the functional versatility of T-cell receptor

Lipid-dependent conformational dynamics underlie the functional versatility of T-cell receptor

bisFabs: Tools for rapidly screening hybridoma IgGs for their activities as bispecific antibodies

Surface expression of the hRSV nucleoprotein impairs immunological synapse formation with T cells

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