Itk functions to control actin polymerization at the immune synapse through localized activation of Cdc42 and WASP

Christine M. Labno; Carol M. Lewis; Daoqi You; Daisy W. Leung; Ana Takesono; Natalie Kamberos; Abhinav Seth; Lisa D. Finkelstein; Michael K. Rosen; Pamela L. Schwartzberg; Janis K. Burkhardt

doi:10.1016/j.cub.2003.08.005

Itk functions to control actin polymerization at the immune synapse through localized activation of Cdc42 and WASP

Christine M. Labno, Carol M. Lewis, Daoqi You, Daisy W. Leung, Ana Takesono, Natalie Kamberos, Abhinav Seth, Lisa D. Finkelstein, Michael K. Rosen, Pamela L. Schwartzberg, Janis K. Burkhardt

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111 Scopus citations

Abstract

Actin polymerization at the immune synapse is required for T cell activation and effector function; however, the relevant regulatory pathways remain poorly understood. We showed previously that binding to antigen presenting cells (APCs) induces localized activation of Cdc42 and Wiskott-Aldrich Syndrome protein (WASP) at the immune synapse [1]. Several lines of evidence suggest that Tec kinases could interact with WASP-dependent actin regulatory processes [2]. Since T cells from RIk-/-, Itk-/-, and RIk-/- x Itk-/- mice have defects in signaling and development [3], we asked whether Itk or RIk function in actin polymerization at the immune synapse. We find that Itk-/- and RIk-/- x Itk-/- T cells are defective in actin polymerization and conjugate formation in response to antigen-pulsed APCs. Itk functions downstream of the TCR, since similar defects were observed upon TCR engagement alone. Using conformation-specific probes, we show that although the recruitment of WASP and Arp2/3 complex to the immune synapse proceeds normally, the localized activation of Cdc42 and WASP is defective. Finally, we find that the defect in Cdc42 activation likely stems from a requirement for Itk in the recruitment of Vav to the immune synapse. Our results identify Itk as a key element of the pathway leading to localized actin polymerization at the immune synapse.

Original language	English (US)
Pages (from-to)	1619-1624
Number of pages	6
Journal	Current Biology
Volume	13
Issue number	18
DOIs	https://doi.org/10.1016/j.cub.2003.08.005
State	Published - Sep 16 2003

ASJC Scopus subject areas

Neuroscience(all)
Biochemistry, Genetics and Molecular Biology(all)
Agricultural and Biological Sciences(all)

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10.1016/j.cub.2003.08.005

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@article{9081d76226344ef2973ff29b38201465,

title = "Itk functions to control actin polymerization at the immune synapse through localized activation of Cdc42 and WASP",

abstract = "Actin polymerization at the immune synapse is required for T cell activation and effector function; however, the relevant regulatory pathways remain poorly understood. We showed previously that binding to antigen presenting cells (APCs) induces localized activation of Cdc42 and Wiskott-Aldrich Syndrome protein (WASP) at the immune synapse [1]. Several lines of evidence suggest that Tec kinases could interact with WASP-dependent actin regulatory processes [2]. Since T cells from RIk-/-, Itk-/-, and RIk-/- x Itk-/- mice have defects in signaling and development [3], we asked whether Itk or RIk function in actin polymerization at the immune synapse. We find that Itk-/- and RIk-/- x Itk-/- T cells are defective in actin polymerization and conjugate formation in response to antigen-pulsed APCs. Itk functions downstream of the TCR, since similar defects were observed upon TCR engagement alone. Using conformation-specific probes, we show that although the recruitment of WASP and Arp2/3 complex to the immune synapse proceeds normally, the localized activation of Cdc42 and WASP is defective. Finally, we find that the defect in Cdc42 activation likely stems from a requirement for Itk in the recruitment of Vav to the immune synapse. Our results identify Itk as a key element of the pathway leading to localized actin polymerization at the immune synapse.",

author = "Labno, {Christine M.} and Lewis, {Carol M.} and Daoqi You and Leung, {Daisy W.} and Ana Takesono and Natalie Kamberos and Abhinav Seth and Finkelstein, {Lisa D.} and Rosen, {Michael K.} and Schwartzberg, {Pamela L.} and Burkhardt, {Janis K.}",

note = "Funding Information: We thank Anne Sperling and Matt Welch for gifts of antibodies and Frances Weis-Garcia and the Monoclonal Antibody Core Facility of Memorial Sloan-Kettering Cancer Center for their technical assistance in generating mAbs. We thank Shirley Bond for assistance with microscopy, Ana Venegas for technical assistance with mice, Jill Voss and Amy Dodson-Watts for administrative assistance, and members of the Burkhardt, Rosen, and Schwartzberg laboratories for many helpful discussions. Peptide synthesis, flow cytometry, antibody purification, and fluorescence microscopy were performed by using the University of Chicago Cancer Research Center Core Facilities. This research was supported by National Institutes of Health grants R01-AI44835 to J.K.B. and R01-GM56322 and Welch grant I-51544 to M.K.R. P.L.S. was supported in part by the Searle Scholars program/Chicago Community Trust, C.M. Lewis was supported by the Howard Hughes Medical Institute-National Institutes of Health Scholars Program, A.T. was supported by the Japan Society for the Promotion of Science, and L.D.F. was supported by the American Cancer Society.",

year = "2003",

month = sep,

day = "16",

doi = "10.1016/j.cub.2003.08.005",

language = "English (US)",

volume = "13",

pages = "1619--1624",

journal = "Current Biology",

issn = "0960-9822",

publisher = "Cell Press",

number = "18",

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TY - JOUR

T1 - Itk functions to control actin polymerization at the immune synapse through localized activation of Cdc42 and WASP

AU - Labno, Christine M.

AU - Lewis, Carol M.

AU - You, Daoqi

AU - Leung, Daisy W.

AU - Takesono, Ana

AU - Kamberos, Natalie

AU - Seth, Abhinav

AU - Finkelstein, Lisa D.

AU - Rosen, Michael K.

AU - Schwartzberg, Pamela L.

AU - Burkhardt, Janis K.

N1 - Funding Information: We thank Anne Sperling and Matt Welch for gifts of antibodies and Frances Weis-Garcia and the Monoclonal Antibody Core Facility of Memorial Sloan-Kettering Cancer Center for their technical assistance in generating mAbs. We thank Shirley Bond for assistance with microscopy, Ana Venegas for technical assistance with mice, Jill Voss and Amy Dodson-Watts for administrative assistance, and members of the Burkhardt, Rosen, and Schwartzberg laboratories for many helpful discussions. Peptide synthesis, flow cytometry, antibody purification, and fluorescence microscopy were performed by using the University of Chicago Cancer Research Center Core Facilities. This research was supported by National Institutes of Health grants R01-AI44835 to J.K.B. and R01-GM56322 and Welch grant I-51544 to M.K.R. P.L.S. was supported in part by the Searle Scholars program/Chicago Community Trust, C.M. Lewis was supported by the Howard Hughes Medical Institute-National Institutes of Health Scholars Program, A.T. was supported by the Japan Society for the Promotion of Science, and L.D.F. was supported by the American Cancer Society.

PY - 2003/9/16

Y1 - 2003/9/16

N2 - Actin polymerization at the immune synapse is required for T cell activation and effector function; however, the relevant regulatory pathways remain poorly understood. We showed previously that binding to antigen presenting cells (APCs) induces localized activation of Cdc42 and Wiskott-Aldrich Syndrome protein (WASP) at the immune synapse [1]. Several lines of evidence suggest that Tec kinases could interact with WASP-dependent actin regulatory processes [2]. Since T cells from RIk-/-, Itk-/-, and RIk-/- x Itk-/- mice have defects in signaling and development [3], we asked whether Itk or RIk function in actin polymerization at the immune synapse. We find that Itk-/- and RIk-/- x Itk-/- T cells are defective in actin polymerization and conjugate formation in response to antigen-pulsed APCs. Itk functions downstream of the TCR, since similar defects were observed upon TCR engagement alone. Using conformation-specific probes, we show that although the recruitment of WASP and Arp2/3 complex to the immune synapse proceeds normally, the localized activation of Cdc42 and WASP is defective. Finally, we find that the defect in Cdc42 activation likely stems from a requirement for Itk in the recruitment of Vav to the immune synapse. Our results identify Itk as a key element of the pathway leading to localized actin polymerization at the immune synapse.

AB - Actin polymerization at the immune synapse is required for T cell activation and effector function; however, the relevant regulatory pathways remain poorly understood. We showed previously that binding to antigen presenting cells (APCs) induces localized activation of Cdc42 and Wiskott-Aldrich Syndrome protein (WASP) at the immune synapse [1]. Several lines of evidence suggest that Tec kinases could interact with WASP-dependent actin regulatory processes [2]. Since T cells from RIk-/-, Itk-/-, and RIk-/- x Itk-/- mice have defects in signaling and development [3], we asked whether Itk or RIk function in actin polymerization at the immune synapse. We find that Itk-/- and RIk-/- x Itk-/- T cells are defective in actin polymerization and conjugate formation in response to antigen-pulsed APCs. Itk functions downstream of the TCR, since similar defects were observed upon TCR engagement alone. Using conformation-specific probes, we show that although the recruitment of WASP and Arp2/3 complex to the immune synapse proceeds normally, the localized activation of Cdc42 and WASP is defective. Finally, we find that the defect in Cdc42 activation likely stems from a requirement for Itk in the recruitment of Vav to the immune synapse. Our results identify Itk as a key element of the pathway leading to localized actin polymerization at the immune synapse.

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ER -

Itk functions to control actin polymerization at the immune synapse through localized activation of Cdc42 and WASP

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