Stromal cell-derived factor 1-α (SDF)-induced human T cell chemotaxis becomes phosphoinositide 3-kinase (PI3K)-independent: Role of PKC-θ

Nahid A. Shahabi, K. McAllen, Burt Sharp

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Stromal cell-derived factor 1α (SDF-1α) is the exclusive ligand for the chemokine receptor CXCR4. This receptor plays a pivotal role in immune responses, the pathogenesis of infection such as HIV, and cellular trafficking. However, the signaling mechanisms regulating SDF-driven T cell migration are not well defined. In this study, we determined the role of PI3K and protein kinase C- θ (PKC-θ) in SDF-induced human T cell migration in fresh versus cultured T cells. Purified human T cells (fresh vs. 48 h in media, unstimulated or activated by anti-CD3+anti-CD28) were used. Western blots showed that SDF induced phospho-(p)-Akt [threonine (Thr)308 and serine 473], a proxy for PI3K activity, in fresh cells and p-PKC-θ in 48 h unstimulated cells. LY294002 (PI3K inhibitor) reduced SDF-induced chemotaxis in fresh cells by 51%, whereas it minimally affected chemotaxis in 48 h unstimulated or activated cells. However, a specific PKC-θ inhibitor, pseudosubstrate for PKC-θ, reduced chemotaxis in 48 h unstimulated and stimulated T cells by 72% and 87%, respectively. Thus, chemotaxis becomes independent of PI3K signaling in human T cells cultured for 48 h. Under these conditions, PKC-θ is phosphorylated (Thr538) by SDF, and chemotaxis becomes largely PKC-θ-dependent.

Original languageEnglish (US)
Pages (from-to)663-671
Number of pages9
JournalJournal of Leukocyte Biology
Volume83
Issue number3
DOIs
StatePublished - Mar 1 2008

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Chemokine CXCL12
1-Phosphatidylinositol 4-Kinase
Chemotaxis
Protein Kinase C
T-Lymphocytes
Cell Movement
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Protein C Inhibitor
Chemokine Receptors
Proxy
Threonine
Protein Kinase Inhibitors
Serine
Cultured Cells
Western Blotting
HIV
Ligands
Infection

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology
  • Cell Biology

Cite this

Stromal cell-derived factor 1-α (SDF)-induced human T cell chemotaxis becomes phosphoinositide 3-kinase (PI3K)-independent : Role of PKC-θ. / Shahabi, Nahid A.; McAllen, K.; Sharp, Burt.

In: Journal of Leukocyte Biology, Vol. 83, No. 3, 01.03.2008, p. 663-671.

Research output: Contribution to journalArticle

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abstract = "Stromal cell-derived factor 1α (SDF-1α) is the exclusive ligand for the chemokine receptor CXCR4. This receptor plays a pivotal role in immune responses, the pathogenesis of infection such as HIV, and cellular trafficking. However, the signaling mechanisms regulating SDF-driven T cell migration are not well defined. In this study, we determined the role of PI3K and protein kinase C- θ (PKC-θ) in SDF-induced human T cell migration in fresh versus cultured T cells. Purified human T cells (fresh vs. 48 h in media, unstimulated or activated by anti-CD3+anti-CD28) were used. Western blots showed that SDF induced phospho-(p)-Akt [threonine (Thr)308 and serine 473], a proxy for PI3K activity, in fresh cells and p-PKC-θ in 48 h unstimulated cells. LY294002 (PI3K inhibitor) reduced SDF-induced chemotaxis in fresh cells by 51{\%}, whereas it minimally affected chemotaxis in 48 h unstimulated or activated cells. However, a specific PKC-θ inhibitor, pseudosubstrate for PKC-θ, reduced chemotaxis in 48 h unstimulated and stimulated T cells by 72{\%} and 87{\%}, respectively. Thus, chemotaxis becomes independent of PI3K signaling in human T cells cultured for 48 h. Under these conditions, PKC-θ is phosphorylated (Thr538) by SDF, and chemotaxis becomes largely PKC-θ-dependent.",
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