SOCS3与LIGHT在诱导DC分化成熟中的相互作用  

Interaction Between Member LIGHT of TNF Superfamily and SOCS3,which Respond to Induce the Differentiation and Maturation of Dendritic Cell

作　　者：林苹[1] 张洁[1] 王琪[1] 陆燕蓉[2] 王修杰[1] 熊竹娟[1] 杨洪亮[1] 任婧婧[1] 

机构地区：[1]四川大学华西医院生物治疗国家重点实验室实验肿瘤研究室,成都610041 [2]四川大学华西医院卫生部移植工程与移植免疫重点实验室,成都610041

出　　处：《四川大学学报（医学版）》2007年第4期644-648,共5页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号3980014730270588)资助

摘　　要：目的探讨SOCS3与LIGHT在刺激树突状细胞(DC)分化成熟过程中的相互作用。方法用重组小鼠粒-巨噬细胞集落刺激因子(rmGM-CSF)、重组小鼠白细胞介素4(rmIL-4)体外诱导培养小鼠骨髓单核细胞产生骨髓来源树突状细胞(BMDC)。RT-PCR、Western blot检测受LIGHT刺激后BMDC中SOCS3 mRNA及SOCS3蛋白表达的时间动力学;利用反义核苷酸技术抑制SOCS3的表达,通过流式细胞术测定BMDC表面分子CD40、CD86的表达水平。结果LIGHT刺激后,BMDC的SOCS3 mRNA水平有不同程度增高(P<0.05),并呈双峰度变化(分别出现在LIGHT刺激后2h和10h,增高幅度分别为92%和83%,其蛋白表达亦有所增加(LIGHT刺激24h后表达量增加80%,P<0.05);反义寡核苷酸能阻断SOCS3的表达,其对mRNA和蛋白表达的抑制率分别为49%和45%;经反义寡核苷酸处理后的BMDC在LIGHT刺激下其CD40、CD86表达均升高(P<0.05)。结论LIGHT在刺激BMDC分化成熟过程中同时上调了SOCS3的表达;阻断SOCS3能使BMDC对LIGHT的反应更敏感,从而促使BMDC成熟度更高;SOCS3对LIGHT诱导的BMDC分化成熟有负反馈调节作用。Objective To detect the relation between the member LIGHT of TNF superfamily and the suppressor of cytokine signaling 3 （SOCS3）, and to investigate the effect of SOCS3 on dendritic cell （DC） maturation induced by LIGHT. Methods Bone marrow-derived DC （BMDC） was generated from mouse bone marrow monocyte by culturing with rmGM-CSF,rmIL-4 in vitro. SOCS3 mRNA in BMDC was analyzed by RT- PCR, and the protein of SOCS3 was measured by Western blot. After blocking the SOCS3 expression with the specific anti-sense oligonucleotide, we applied the flow cytometry to measure the expression of CD86 and CD40 on DC for making clear whether the silence of SOCS3 would regulate the LIGHT-stimulated DC maturation. Results With the effect of LIGHT, the level of SOCS3 mRNA and protein in BMDC sharply increased. The specific antisense oligonucleotide could effectively block SOCS3 mRNA expressing in BMDC with the ratio of 49% and block SOCS3 protein expression with the ratio of 45%. Compared with SOCS3-unblocked DC, the SOCS3-blocked BMDC with stimulation of LIGHT showed higher CD40 and CD86 （P〈0. 05）. Conclusion LIGHT enhances the expression of SOCS3 during stimulating BMDC maturation. As more sensitive to LIGHT, the SOCS3-blocked BMDC is driven to more mature. SOCS3 presents a negative regulation mechanism in BMDC maturation induced by LIGHT.

关 键 词：LIGHT SOCS3 树突状细胞 分化成熟 

分 类 号：R392.12[医药卫生—免疫学]