转录因子Hlx修饰树突状细胞系DC2.4的功能研究  

Functional study of transcription factor Hlx modified dendritic cell line DC2.4

作　　者：周成林[1,2] 苏兆亮[1,2] 朱金连[1] 何志强[2] 王胜军[2] 李平[1] 许化溪[2] 

机构地区：[1]镇江出入境检验检疫局,江苏镇江212008 [2]江苏大学检验医学研究所免疫研究室,江苏镇江212013

出　　处：《细胞与分子免疫学杂志》2011年第3期274-277,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30871193;81001319);江苏省高校自然基金资助项目(09KJB310001);江苏省高校博士创新基金资助项目(CX09B_217Z)

摘　　要：目的:在鼠树突状细胞系DC2.4中瞬时表达Th1特异的核转录Hlx,探讨该转录因子对树突状细胞系DC2.4的功能的影响。方法:含有Hlx基因全部编码序列的真核表达载体PIRES2-EGFP/Hlx通过脂质体转染的方法转入DC细胞,转染效率应用FACS鉴定;Hlx基因在DC中表达情况应用RT-PCR和Real-time PCR检测。转染48 h后,分别从细胞因子,表面分子,吞噬功能,单向混合淋巴细胞反应方面对DC功能进行探讨。结果:真核表达载体PIRES2-EGFP/Hlx成功转入DC细胞,转染率可高达60%;在树突状细胞系DC2.4中瞬时高表达Hlx可以有效的增进成熟标志CD80,CD86,MHCII分子的表达,减弱DC的吞噬功能,IL-12表达量增加,但同时却高表达IL-10,TGF-β并表现为可抑制淋巴细胞增殖。结论:在小鼠树突状细胞系DC2.4中瞬时高表达Hlx可有效地促进DC2.4的成熟,但是在功能方面多表现为调节性DC的功能。AIM： To transfect Hlx into mouse dendritic cell line DC2.4 and observe the effect of hlx on function of dendritic cells.METHODS： The eukaryotic expression vector PIRES2-EGFP/Hlx was transfected into DC2.4 by liposomes.The transfection efficiency was identified through FACS.RT-PCR and Real-time PCR were used to test the transcription level of Hlx in DC2.4.Forty-eight hours after transfection,DC2.4 cells were studied for cytokine production,cell phenotype,phagocytosis,unilateral mixed lymphocyte reaction.RESULTS： The pIRES2-EGFP/Hlx vector was transfected into DC2.4 with the transfection efficiency of up to 60%.Highly expressed Hlx in DC2.4 increased the expression of maturation makers including CD80 and CD86,and major histocompatibility complex-II.Functional assay showed that over-expression of Hlx in DC2.4 increased the interleukin-12 transcription and decreased DC endocytosis.The Hlx modified DC2.4 highly expressed IL-10 and TGF-β at the same time.Furthermore,it was shown that in a unilateral mixed lymphocyte reaction model,Hlx modified DC2.4 inhibited proliferation of lymphocytes.CONCLUSION： Transient over-expression of Hlx in DC2.4 promotes DC2.4 maturation and up-regulates IL-12,IL-10 and TGF-β expression.However,the Hlx modified DC2.4 cells functionally appear as regulatory dendritic cells.

关 键 词：IL-10 Hlx 树突状细胞 调节性DC 

分 类 号：R446.63[医药卫生—诊断学]