Small Interfering RNAs Specific for Spleen Tyrosine Kinase InhibitMaturation of Dendritic Cells of AsthmaticMice in Vitro_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

ZHENG Yonghua ¹ , LU Jingjing ² , GUO Zhongliang ² , REN Tao ² ,  LIANG Yongjie. ²

Department of Respiratory Medicine, Dongyang People’s Hospital. Dongyang, Zhejiang, 322100, China Corresponding Author: LIANG Yong-jie, E-mail: liangyj2008@ yahoo. com. cn;

Corresponding author：

LIANG Yongjie., Email: liangyj2008@yahoo.com.cn

Keywords：

Asthma; Small interfering RNA; Dendritic cells; Spleen tyrosine kinase; Cytokines

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Objective By using small interfering RNAs ( siRNAs) specific for spleen tyrosine kinase ( Syk) , to evaluate the role of Syk in maturation of bone marrow-derived dendritic cells. Methods The fragments of 21-23 bp siRNAs specific for mice Syk were chemo synthesized and transfected into the asthmatic murine bone marrow-derived dendritic cells ( BMDCs) by Lipofectamine 2000 transfection system for 48 hours. Then BMDCs were co-cultured with T cells from the normal mice spleen for 48 hours. The cytokines including IL-4, IL-13, IL-2 and INF-γin supernatant were detect by ELISA. The expression of Syk protein was measured by Western Blot to determine whether the Syk gene was silenced. Results The expression of Syk protein was obviously decreased in the siRNA-interference group. The secretions of IL-4 and IL-13 were significantly inhibited by siRNA interference ( P lt; 0. 05) , but the secretions of IL-2 and INF-γwere not interfered signficantly ( P gt;0. 05) . Conclusion Syk specific siRNA fragments can block the antigen presentation function of dendritic cells and block the activation and differentiation of T cells.

Citation： ZHENG Yonghua,LU Jingjing,GUO Zhongliang,REN Tao,LIANG Yongjie.. Small Interfering RNAs Specific for Spleen Tyrosine Kinase InhibitMaturation of Dendritic Cells of AsthmaticMice in Vitro. Chinese Journal of Respiratory and Critical Care Medicine, 2009, 09(5): 487-491. doi: Copy

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1. Elias JA, Zhu Z, Chupp G, et al. Airway remodeling in asthma. J Clin Invest, 1999, 104: 1001-1006.
2. Elias JA, Lee CG, Tao Zet al. New insights into the pathogenesis of ·490· 中国呼吸与危重监护杂志2009 年9 月第8 卷第5 期Chin J Respir Crit Care Med, September 2009 , Vol.
3. 马艳良, 何权瀛. 从哮喘的流行病学调查看我国哮喘诊断不足问题. 中国呼吸与危重监护杂志, 2004, 3: 10-14.
4. 刘春涛, 冯玉麟, 梁宗安, 等. 支气管哮喘防治现状与对策的卫生经济学分析. 中国呼吸与危重监护杂志, 2002, 1: 148-150.
5. 钟南山, 主编. 支气管哮喘与临床. 北京: 人民卫生出版社, 2006, 450-451.
7. Guermonprez P, Valladeau J, Zitvogel L, et al. Antigen presentation and T cell stimulation by dendritic cells. Annu Rev Immunol, 2002 , 20: 621-667.
8. Hannon GJ. RNA interference. Nature, 2002, 418: 244-251.
9. Plasterk RHA. RNA Silencing: The Genome’s Immune System. Science, 2002, 296: 1263-1265.
10. , No.
11. Wong WS, Leong KP. Tyrosine kinase inhibitors: a new approach for asthma. Biochim Biophys Acta, 2004, 1697: 53-69.
12. 程晓明, 王长征, 李淑平, 等. 哮喘致敏小鼠模型的建立及脾脏 CD4 + T 细胞的分离. 第三军医大学学报, 2001, 23: 369-370.
13. Cohn L, Elias JA, Chupp GL. Asthma: mechanisms of disease persistence and progression. Annu Rev Immunol, 2004 , 22: 789 - 815.
14. Nadler MJ, Matthews SA, Turner H, et al. Signal transduction by the high-affinity immunoglobulin E receptor FcqRI: coupling form to function. Adv Immunol, 2000, 76: 325-355.
15. Nel AE. T-cell activation through the antigen receptor. Part 1 : Signaling components, signaling pathways, and signal integration at the T-cell antigen receptor synapse. J Allergy Clin Immunol, 2002 , 109: 758-770.
16. Samelson LE. Signal transduction mediated by the T cell antigen receptor: the role of adapter proteins. Annu Rev Immunol, 2002, 20: 371-394.
17. Gauld SB, Dal Porto JM, Cambier JC. B cell antigen receptor signaling: roles in cell development and disease. Science, 2002 , 296: 1641-1642.
18. Hong JJ, Yankee TM, Harrison ML, et al. Regulation of signaling in B cells through the phosphorylation of Syk on linker region tyrosines. J Biol Chem, 2002, 277: 31703-31714.
19. Matsubara S, Koya T, Takeda K, et al. Syk activation in dendritic cells is essential for airway hyperresponsiveness and inflammation. Am J Respir Cell Mol Biol, 2006, 34: 426-433.
20. emann UA, Ray P, FlaveII RA. Pulmonary overexpression of IL-9 induces Th2 cytokine expression, leading to immune pathology. J Clin Invest, 2002, 109: 29-39.
21. 毛光宇, 杨炯, 陈宏斌, 等. 过敏性哮喘患者树突状细胞对原始T 细胞活化的影响. 中国呼吸与危重监护杂志, 2005, 4: 115-118.
22. Ma B, Zhu Z, Homer RJ, et al. The C10 /CCL6 chemokine and CCR1 play critical roles in the pathogenesis of IL-13-induced inflammation and remodeling. J Immunol, 2004, 172: 1872-1881.
23. Zhu Z, Ma B, Zheng T, et al. IL-13-induced chemokine responses in the lung: Role of CCR2 in the pathogenesis of IL-13-induced inflammation and remodeling. J Immunol, 2002, 168: 2953-2962.
24. Prechtel AT, Turza NM, Theodoridis AA, et al. Small interfering RNA ( siRNA ) delivery into monocyte-derived dendritic cells by electroporation. J Immunol Methods, 2006, 311: 139-152.

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