Spatial and Temporal Dynamics of T Cell Receptor Signaling with a Photoactivatable Agonist

Morgan Huse; Lawrence O. Klein; Andrew T. Girvin; Joycelyn M. Faraj; Qi Jing Li; Michael S. Kuhns; Mark M. Davis

doi:10.1016/j.immuni.2007.05.017

Spatial and Temporal Dynamics of T Cell Receptor Signaling with a Photoactivatable Agonist

Morgan Huse, Lawrence O. Klein, Andrew T. Girvin, Joycelyn M. Faraj, Qi Jing Li, Michael S. Kuhns, Mark M. Davis

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

194 Scopus citations

Abstract

The precise timing of signals downstream of the T cell receptor (TCR) is poorly understood. To address this problem, we prepared major histocompatibility complexes containing an antigenic peptide that is biologically inert until exposed to ultraviolet (UV) light. UV irradiation of these complexes in contact with cognate T cells enabled the high-resolution temporal analysis of signaling. Phosphorylation of the LAT adaptor molecule was observed in 4 s, and diacylglycerol production and calcium flux was observed in 6-7 s. TCR activation also induced cytoskeletal polarization within 2 min. Antibody blockade of CD4 reduced the intensity of LAT phosphorylation and the speed of calcium flux. Furthermore, strong desensitization of diacylglycerol production, but not LAT phosphorylation, occurred shortly after TCR activation, suggesting that different molecular events play distinct signal-processing roles. These results establish the speed and localization of early signaling steps, and have important implications regarding the overall structure of the network.

Original language	English (US)
Pages (from-to)	76-88
Number of pages	13
Journal	Immunity
Volume	27
Issue number	1
DOIs	https://doi.org/10.1016/j.immuni.2007.05.017
State	Published - Jul 27 2007
Externally published	Yes

Keywords

MOLIMMUNO

ASJC Scopus subject areas

Immunology and Allergy
Immunology
Infectious Diseases

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10.1016/j.immuni.2007.05.017

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@article{b6eba40bc79c416b9d9f1a594b431bed,

title = "Spatial and Temporal Dynamics of T Cell Receptor Signaling with a Photoactivatable Agonist",

abstract = "The precise timing of signals downstream of the T cell receptor (TCR) is poorly understood. To address this problem, we prepared major histocompatibility complexes containing an antigenic peptide that is biologically inert until exposed to ultraviolet (UV) light. UV irradiation of these complexes in contact with cognate T cells enabled the high-resolution temporal analysis of signaling. Phosphorylation of the LAT adaptor molecule was observed in 4 s, and diacylglycerol production and calcium flux was observed in 6-7 s. TCR activation also induced cytoskeletal polarization within 2 min. Antibody blockade of CD4 reduced the intensity of LAT phosphorylation and the speed of calcium flux. Furthermore, strong desensitization of diacylglycerol production, but not LAT phosphorylation, occurred shortly after TCR activation, suggesting that different molecular events play distinct signal-processing roles. These results establish the speed and localization of early signaling steps, and have important implications regarding the overall structure of the network.",

keywords = "MOLIMMUNO",

author = "Morgan Huse and Klein, {Lawrence O.} and Girvin, {Andrew T.} and Faraj, {Joycelyn M.} and Li, {Qi Jing} and Kuhns, {Michael S.} and Davis, {Mark M.}",

note = "Funding Information: We thank M. Bose and A. Sanchez for assistance with chemical synthesis and G. Marriott for advice on photoactivation. We also thank Q. Hu for help with CD4-blocking studies, J. Huppa for the α-tubulin-YFP retrovirus, N. Prado for technical support, and other members of the Davis lab for helpful discussions. M.H. was supported by a Giannini Family Foundation Fellowship, L.O.K. was supported by a National Defense Science and Engineering Graduate Fellowship, A.T.G. was supported by a National Science Foundation Graduate Fellowship, J.M.F. was supported by the Stanford Summer Research Program in Biomedical Sciences, and M.S.K. was supported by a senior fellowship from the Irvington Institute for Immunological Research. Additional support was provided by grants from the National Institutes of Health and from the Howard Hughes Medical Institute to M.M.D. ",

year = "2007",

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doi = "10.1016/j.immuni.2007.05.017",

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T1 - Spatial and Temporal Dynamics of T Cell Receptor Signaling with a Photoactivatable Agonist

AU - Huse, Morgan

AU - Klein, Lawrence O.

AU - Girvin, Andrew T.

AU - Faraj, Joycelyn M.

AU - Li, Qi Jing

AU - Kuhns, Michael S.

AU - Davis, Mark M.

N1 - Funding Information: We thank M. Bose and A. Sanchez for assistance with chemical synthesis and G. Marriott for advice on photoactivation. We also thank Q. Hu for help with CD4-blocking studies, J. Huppa for the α-tubulin-YFP retrovirus, N. Prado for technical support, and other members of the Davis lab for helpful discussions. M.H. was supported by a Giannini Family Foundation Fellowship, L.O.K. was supported by a National Defense Science and Engineering Graduate Fellowship, A.T.G. was supported by a National Science Foundation Graduate Fellowship, J.M.F. was supported by the Stanford Summer Research Program in Biomedical Sciences, and M.S.K. was supported by a senior fellowship from the Irvington Institute for Immunological Research. Additional support was provided by grants from the National Institutes of Health and from the Howard Hughes Medical Institute to M.M.D.

PY - 2007/7/27

Y1 - 2007/7/27

N2 - The precise timing of signals downstream of the T cell receptor (TCR) is poorly understood. To address this problem, we prepared major histocompatibility complexes containing an antigenic peptide that is biologically inert until exposed to ultraviolet (UV) light. UV irradiation of these complexes in contact with cognate T cells enabled the high-resolution temporal analysis of signaling. Phosphorylation of the LAT adaptor molecule was observed in 4 s, and diacylglycerol production and calcium flux was observed in 6-7 s. TCR activation also induced cytoskeletal polarization within 2 min. Antibody blockade of CD4 reduced the intensity of LAT phosphorylation and the speed of calcium flux. Furthermore, strong desensitization of diacylglycerol production, but not LAT phosphorylation, occurred shortly after TCR activation, suggesting that different molecular events play distinct signal-processing roles. These results establish the speed and localization of early signaling steps, and have important implications regarding the overall structure of the network.

AB - The precise timing of signals downstream of the T cell receptor (TCR) is poorly understood. To address this problem, we prepared major histocompatibility complexes containing an antigenic peptide that is biologically inert until exposed to ultraviolet (UV) light. UV irradiation of these complexes in contact with cognate T cells enabled the high-resolution temporal analysis of signaling. Phosphorylation of the LAT adaptor molecule was observed in 4 s, and diacylglycerol production and calcium flux was observed in 6-7 s. TCR activation also induced cytoskeletal polarization within 2 min. Antibody blockade of CD4 reduced the intensity of LAT phosphorylation and the speed of calcium flux. Furthermore, strong desensitization of diacylglycerol production, but not LAT phosphorylation, occurred shortly after TCR activation, suggesting that different molecular events play distinct signal-processing roles. These results establish the speed and localization of early signaling steps, and have important implications regarding the overall structure of the network.

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Spatial and Temporal Dynamics of T Cell Receptor Signaling with a Photoactivatable Agonist

Abstract

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