CⅡTA反义RNA抑制皮肤成纤维细胞表面MHCⅡ类抗原表达  

Inhibition of MHCⅡExpression on Human Dermal Fibroblast Through CⅡTA Anti-sense RNA

作　　者：郭荣[1,2] 邹萍[1] 陆华中[3] 范华骅[3] 曹谊林[4] 崔磊[4] 刘伟[4] 商庆新[4] 郑滨[3] 高跞[3] 高峰[3] 

机构地区：[1]华中科技大学同济医学院附属协和医院血研所 [2]广东省人民医院血液科,510080 [3]上海市血液中心血液工程研究室 [4]上海第二医科大学组织工程研究重点实验室

出　　处：《中华皮肤科杂志》2003年第11期619-622,共4页Chinese Journal of Dermatology

基　　金：上海市科技发展基金资助项目(0143nm068);上海市科技发展基金/重大项目子课题资助项目(00DJ14001-8)

摘　　要：目的探讨用纳米载体介导主要组织相容性复合物(majorhistocompatibilitycomplex,MHC)Ⅱ类抗原转录激活因子(MHCⅡtransactivator,CⅡTA)的反义RNA,抑制皮肤成纤维细胞表面MHCⅡ类抗原表达的可行性。方法将CⅡTA反义RNA(pDarⅡ质粒)稳定转染人类原代皮肤成纤维细胞(pDarⅡ-D),流式细胞仪检测pDarⅡ-D表面经典的MHCⅡ(HLA-DR、-DP、-DQ)类抗原表达,RT-PCR检测CⅡTA及经典的MHCⅡ类分子的mRNA水平。体外混合淋巴细胞反应检测pDarⅡ-D组刺激外周血T细胞反应的能力。结果与正义链组比较,在重组人干扰素(IFN)-γ诱导下,pDarⅡ-D组HLA-DR及-DP抗原诱导性表达分别降低了95.63%、87.89%;同时CⅡTA及经典的MHCⅡ类分子的mRNA含量明显减少(P<0.01);pDarⅡ-D组刺激T细胞分泌IL-2mRNA水平降低(P<0.05)。结论CⅡTA反义片段抑制了自身mRNA含量,从而阻止了其调控的MHCⅡ类分子的相应表达。Objective To investigate the feasibility of using anti-sense RNA against classⅡmajor histocompatibility complex(MHCⅡ)transactivator(CⅡTA),which might regulate MHCⅡexpression,to suppress the relative immune response.Methods Stable transfectants of dermal fibroblasts with pDarⅡ(pDarⅡ-D)were tested for the expression of classic MHCⅡ(HLA-DR,-DP,-DQ)antigens induced with recombinant human interferon-gamma(IFN-γ).mRNA abundance of CⅡTA,and classic MHCⅡwas mea-sured by RT-PCR.IL-2mRNA expressed in T cells,stimulated by transfected dermal fibroblasts,was de-termined by mixed lymphocyte reaction.Results When induced with IFN-γ,the expression of HLA-DR and-DP antigens on pDarⅡ-D was reduced by95.63%and87.89%,respectively.Meanwhile,the mRNA contents of CⅡTA and classic MHCⅡwere decreased significantly(P<0.01).T cells expressed less IL-2mRNA in the presence of pDarⅡ-D.Conclusion pDarⅡinhibits CⅡTA mRNA and further inhibits its regulation of MHCⅡmolecule expression.

关 键 词：主要组织相容性复合物 II类抗原转录激活因子 皮肤成纤维细胞 同种异体细胞移植 反义RNA 抗原表达 

分 类 号：R392[医药卫生—免疫学]