Inhibition of IL-2 inducible T-cell kinase alleviates T-cell activation and murine myocardial inflammation associated with CVB3 infection

Feng He  ¹ , Hailan Yao  ¹ , Zonghui Xiao  ¹ , Jisheng Han  ² , Jizhen Zou  ³ , Zhewei Liu  ⁴

Affiliations

• ¹ Molecular Immunology Laboratory, Capital Institute of Pediatrics, Beijing 100020, China.
• ² Medical Department, Aerospace 731 Hospital, Beijing 100074, China.
• ³ Pathology Laboratory, Capital Institute of Pediatrics, Beijing 100020, China.
• ⁴ Molecular Immunology Laboratory, Capital Institute of Pediatrics, Beijing 100020, China. Electronic address: zheweiliu@163.com.

Abstract

Background: Coxsackievirus B3 (CVB3) infection causes myocarditis, pancreatitis, and aseptic meningitis. Targeting antigen-specific T cell reactions might be a promising way to alleviate the inflammatory response induced by CVB3 infection. IL-2-inducible T-cell kinase (ITK), a member of Tec kinase family expressed mainly in T cells, plays an important role in the activation of T cells. The role of ITK in viral myocarditis induced by CVB3 has not been documented.

Methodology: In this study, we inhibited the ITK expression in Jurkat cells, primary human peripheral blood mononuclear cells (PBMC), and mouse splenocytes by ITK-specific siRNA. The inhibition efficiently suppressed cell proliferation (P<0.05) and T-cell related cytokine secretion (P<0.05). In order to inhibit ITK in vivo, the pGCSIL plasmid containing short hairpin RNAs targeting ITK was constructed and transduced into mice infected with CVB3. ITK-inhibited mice showed reduced cell proliferation (3, 5, and 7 days post-challenge, P<0.05) as well as CD4+ and CD8+ T cells (5 days post-challenge, P<0.05). The altered production of inflammatory cytokines alleviated pathologic heart damage and improved mice survival rate (P<0.05).

Conclusion: ITK played an important role in the T cell development and represented a new target for the modulation of T-cell-mediated inflammatory response by CVB3 infection.

Keywords: Coxsackievirus B3; ITK; Myocarditis; RNAi.