Hematopoietic lineage cell-specific protein 1 is recruited to the immunological synapse by IL-2-inducible T cell kinase and regulates phospholipase Cγ1 microcluster dynamics during T cell spreading

Esteban Carrizosa, Timothy S. Gomez, Christine M. Labno, Deborah A. Klos Dehring, Xiaohong Liu, Bruce D. Freedman, Daniel D. Billadeau, Janis K. Burkhardt

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

Productive T cell activation requires efficient reorganization of the actin cytoskeleton. We showed previously that the actinregulatory protein, hematopoietic lineage cell-specific protein 1 (HS1), is required for the stabilization of F-actin and Vav1 at the immunological synapse and for efficient calcium responses. The Tec family kinase IL-2-inducible T cell kinase (Itk) regulates similar aspects of T cell activation, suggesting that these proteins act in the same pathway. Using video microscopy, we show that T cells lacking Itk or HS1 exhibited similar defects in actin responses, extending unstable lamellipodial protrusions upon TCR stimulation. HS1 and Itk could be coimmunoprecipitated from T cell lysates, and GST-pulldown studies showed that Itk's Src homology 2 domain binds directly to two phosphotyrosines in HS1. In the absence of Itk, or in T cells overexpressing an Itk Src homology 2 domain mutant, HS1 failed to localize to the immunological synapse, indicating that Itk serves to recruit HS1 to sites of TCR engagement. Because Itk is required for phospholipase C (PLC)γ1 phosphorylation and calcium store release, we examined the calcium signaling pathway in HS1-/- T cells in greater detail. In response to TCR engagement, T cells lacking HS1 exhibited diminished calcium store release, but TCR-dependent PLCγ1 phosphorylation was intact, indicating that HS1's role in calcium signaling is distinct from that of Itk. HS1-deficient T cells exhibited defective cytoskeletal association of PLCγ1 and altered formation of PLCγ1 microclusters. We conclude that HS1 functions as an effector of Itk in the T cell actin-regulatory pathway, and directs the spatial organization of PLCγ1 signaling complexes.

Original languageEnglish (US)
Pages (from-to)7352-7361
Number of pages10
JournalJournal of Immunology
Volume183
Issue number11
DOIs
StatePublished - Dec 1 2009

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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