抗CⅡTA的核糖核酸酶P对Jurkat细胞MHCⅡ类分子表达的抑制作用  

Inhibitory Effect of Anti-CⅡTA RNase P on MHCⅡ Expression in Jurkat Cells

作　　者：何飞[1] 吴书林[1] 孙明[2] 郭荣[3] 

机构地区：[1]广东省人民医院心内科,广东省心血管病研究所,广州510080 [2]中南大学湘雅医院心内科,长沙410078 [3]广东省人民医院血液科,广州510080

出　　处：《中国实验血液学杂志》2007年第3期607-611,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(编号30571771);广东省医学科学技术研究基金资助项目(编号A2005017);广东省人民医院科学技术研究基金资助项目(编号Y2004087);广东省自然科学基金资助项目(编号04003959;编号06020896)

摘　　要：本研究旨在探讨抗MHCⅡ类分子转录激活因子(CⅡTA)的M1-RNA抑制细胞表面MHCⅡ类分子的表达。M1-RNA是核糖核酸酶P的催化活性单位,设计并克隆针对CⅡTA第3408位点的M1-RNA(M1-3408-GS)及其相应的CⅡTA靶基因片段(3176-3560),分别插入pUC19、pGEM-7zf(+)载体,进行细胞外切割活性筛选。将细胞外切割作用明显的M1-3408-GS亚克隆入psNAV载体并稳定转染Jurkat细胞株,采用流式细胞术检测该细胞表面经典的MHCⅡ(HLA-DR、-DP、-DQ)类抗原表达,RT-PCR检测其CⅡTA的mRNA水平。结果表明:在重组人干扰素(IFN)-γ诱导下,M1-3408-GS阳性Jurkat细胞株与对照组比较,其表面HLA-DR、HLA-DP、HLA-DQ抗原诱导型表达分别降低了83.17%、94.12%及84.31%;同时CⅡTA的mRNA含量明显降低(P<0.05,t=4.89)。结论:抗CⅡTA的M1-RNA(M1-3408-GS)降低了自身mRNA含量,从而阻止其调控的MHCⅡ类分子的表达。This study was purposed to investigate the inhibitory effect of anti-C Ⅱ TA M1-RNA on MHC Ⅱ expression. The M1-RNA with guide sequences （GS） recognizing C Ⅱ TA at 3408 site（M1-3408-GS） and C Ⅱ TA target RNA （3176 -3560） were constructed, then cloned into the pUC19 and pGEM-7zf（ ＋ ） vector respectively. The recombinant M1-RNA and its target RNA were incubated in cell-free conditions. It showed that M1-3408-GS could exclusively cleave target RNA, then it was cloned into the psNAV vector. Stable transfectants of Jurkat cells with M1-3408-GS were analyzed for classical MHC Ⅱ （ HLA-DR,-DP,-DQ ） induction in response to IFN-γ by flow cytometry. The level of C Ⅱ TA mRNA was measured by RT-PCR. The results showed that after IFN-γ treatment, the expression of HLA-DR, HLA-DP,HLA-DQ on M1-3408-GS positive Jurkat cells decreased 83.17%, 94.12% and 84.31% respectively as compared with control. At the same time the mRNA contents of C Ⅱ TA also markedly decreased （ P 〈 0.05, t = 4.89）. It is concluded that anti- C Ⅱ TA M1-RNA （ M1-3408-GS ） inhibits C Ⅱ TA, decreases itself mRNA content and so suppresses expression of MHC Ⅱ molecules regulated by C Ⅱ TA.

关 键 词：MHCⅡ类分子转录激活因子(CⅡTA) 核糖核酸酶P(M1-RNA) 移植免疫 

分 类 号：R392.4[医药卫生—免疫学] R733.71[医药卫生—基础医学]