Mechanisms of localized activation of the T cell antigen receptor inside clusters

Yair Neve-Oz; Yair Razvag; Julia Sajman; Eilon Sherman

doi:10.1016/j.bbamcr.2014.09.025

Mechanisms of localized activation of the T cell antigen receptor inside clusters

Yair Neve-Oz, Yair Razvag, Julia Sajman, Eilon Sherman^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

26 Scopus citations

Abstract

The T cell antigen receptor (TCR) has been shown to cluster both before and upon engagement with cognate antigens. However, the effect of TCR clustering on its activation remains poorly understood. Here, we used two-color photo-activated localization microscopy (PALM) to visualize individual molecules of TCR and ZAP-70, as a marker of TCR activation and phosphorylation, at the plasma membrane of uniformly activated T cells. Imaging and second-order statistics revealed that ZAP-70 recruitment and TCR activation localized inside TCR clusters. Live cell PALM imaging showed that the extent of localized TCR activation decreased, yet remained significant, with cell spreading. Using dynamic modeling and Monte-Carlo simulations we evaluated possible mechanisms of localized TCR activation. Our simulations indicate that localized TCR activation is the result of long-range cooperative interactions between activated TCRs, or localized activation by Lck and Fyn. Our results demonstrate the role of molecular clustering in cell signaling and activation, and are relevant to studying a wide range of multi-molecular complexes. This article is part of a Special Issue entitled: Nanoscale membrane organisation and signalling.

Original language	American English
Pages (from-to)	810-821
Number of pages	12
Journal	Biochimica et Biophysica Acta - Molecular Cell Research
Volume	1853
Issue number	4
DOIs	https://doi.org/10.1016/j.bbamcr.2014.09.025
State	Published - 1 Apr 2015

Bibliographical note

Funding Information:
The authors would like to thank Zeiss, Harald Hess (HHMI, Janelia Farm) for providing the PALM software and Thorstan Wiegand (Helmholtz Centre for Environmental Research — UFZ) for providing us his point-pattern analyses software. This research was supported by Grant no. 321993 from the Marie Skłodowska-Curie actions of the European Commission , the Lejwa Fund for Biochemistry , and Grants no. 1417/13 and no. 1937/13 from the Israeli Science Foundation .

Publisher Copyright:
© 2014 Elsevier B.V.

Keywords

Immune synapse
Microcluster
Modeling and simulation
Nanocluster
Photoactivated localization microscopy
Super-resolution light microscopy
T cell activation
T cell receptor

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title = "Mechanisms of localized activation of the T cell antigen receptor inside clusters",

abstract = "The T cell antigen receptor (TCR) has been shown to cluster both before and upon engagement with cognate antigens. However, the effect of TCR clustering on its activation remains poorly understood. Here, we used two-color photo-activated localization microscopy (PALM) to visualize individual molecules of TCR and ZAP-70, as a marker of TCR activation and phosphorylation, at the plasma membrane of uniformly activated T cells. Imaging and second-order statistics revealed that ZAP-70 recruitment and TCR activation localized inside TCR clusters. Live cell PALM imaging showed that the extent of localized TCR activation decreased, yet remained significant, with cell spreading. Using dynamic modeling and Monte-Carlo simulations we evaluated possible mechanisms of localized TCR activation. Our simulations indicate that localized TCR activation is the result of long-range cooperative interactions between activated TCRs, or localized activation by Lck and Fyn. Our results demonstrate the role of molecular clustering in cell signaling and activation, and are relevant to studying a wide range of multi-molecular complexes. This article is part of a Special Issue entitled: Nanoscale membrane organisation and signalling.",

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note = "Funding Information: The authors would like to thank Zeiss, Harald Hess (HHMI, Janelia Farm) for providing the PALM software and Thorstan Wiegand (Helmholtz Centre for Environmental Research — UFZ) for providing us his point-pattern analyses software. This research was supported by Grant no. 321993 from the Marie Sk{\l}odowska-Curie actions of the European Commission , the Lejwa Fund for Biochemistry , and Grants no. 1417/13 and no. 1937/13 from the Israeli Science Foundation . Publisher Copyright: {\textcopyright} 2014 Elsevier B.V.",

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AU - Sherman, Eilon

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N2 - The T cell antigen receptor (TCR) has been shown to cluster both before and upon engagement with cognate antigens. However, the effect of TCR clustering on its activation remains poorly understood. Here, we used two-color photo-activated localization microscopy (PALM) to visualize individual molecules of TCR and ZAP-70, as a marker of TCR activation and phosphorylation, at the plasma membrane of uniformly activated T cells. Imaging and second-order statistics revealed that ZAP-70 recruitment and TCR activation localized inside TCR clusters. Live cell PALM imaging showed that the extent of localized TCR activation decreased, yet remained significant, with cell spreading. Using dynamic modeling and Monte-Carlo simulations we evaluated possible mechanisms of localized TCR activation. Our simulations indicate that localized TCR activation is the result of long-range cooperative interactions between activated TCRs, or localized activation by Lck and Fyn. Our results demonstrate the role of molecular clustering in cell signaling and activation, and are relevant to studying a wide range of multi-molecular complexes. This article is part of a Special Issue entitled: Nanoscale membrane organisation and signalling.

AB - The T cell antigen receptor (TCR) has been shown to cluster both before and upon engagement with cognate antigens. However, the effect of TCR clustering on its activation remains poorly understood. Here, we used two-color photo-activated localization microscopy (PALM) to visualize individual molecules of TCR and ZAP-70, as a marker of TCR activation and phosphorylation, at the plasma membrane of uniformly activated T cells. Imaging and second-order statistics revealed that ZAP-70 recruitment and TCR activation localized inside TCR clusters. Live cell PALM imaging showed that the extent of localized TCR activation decreased, yet remained significant, with cell spreading. Using dynamic modeling and Monte-Carlo simulations we evaluated possible mechanisms of localized TCR activation. Our simulations indicate that localized TCR activation is the result of long-range cooperative interactions between activated TCRs, or localized activation by Lck and Fyn. Our results demonstrate the role of molecular clustering in cell signaling and activation, and are relevant to studying a wide range of multi-molecular complexes. This article is part of a Special Issue entitled: Nanoscale membrane organisation and signalling.

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Mechanisms of localized activation of the T cell antigen receptor inside clusters

Abstract

Bibliographical note

Keywords

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