Ⅱ类抗原反式激活因子及HLADR抗原关系及其意义.docVIP

Ⅱ类抗原反式激活因子及HLADR抗原关系及其意义 　【摘要】 本 研究 探讨Ⅱ类抗原反式激活因子（CIITA）和人类白细胞抗原（HLADR）表达时相的关系和差异，及STAT1α反义寡核苷酸（STAT1α AS）对CIITA和HLADR的抑制作用。分离健康志愿者外周血T淋巴细胞，给予不同剂量干扰素γ（IFNγ）后，用RTPCR法检测CIITA mRNA，Western blot 分析 HLADR抗原表达，然后给予不同浓度STAT1α AS和STAT1α寡核苷酸有义链（STAT1α S），再次检测CIITA mRNA和HLADR的表达。结果表明： CIITA mRNA在IFN γ作用后5小时开始表达，14小时达峰值；HLADR在28小时后可被检测出，52小时达高峰。5、10和20 μmol/L STAT1α AS作用于细胞后，CIITA mRNA的表达显著低于对照组（P＜0.01），而在S组明显高于AS处理组（P＜0.01），S组与对照组间无显著差异；HLADR的表达可被STAT1α AS抑制，AS组仅为对照组的64.3%（P＜0.01），S组与对照组间仍无差异； STAT1α AS作用后，HLADR变化同CIITA。结论： CIITA mRNA表达与HLADR表达呈正相关且早于后者； STAT1α AS可特异性抑制CIITA和HLADR的表达，并能预防T淋巴细胞激活， CIITA在移植免疫病因中起重要作用。 【关键词】 Ⅱ类抗原反式激活因子 HLADR STAT1α 反义寡核苷酸 干扰素γ 毕业论文 This work was supported by National Natural Science Fundation of China（国家 自然 科学 基金）（Noand the 135 project fundation of Jiangsu province The class II transactivator (CIITA) was initially cloned using genetic complementation of the in vitro derived HLADRnegative mutant B cell line, RJ2.2.5［1］. All current evidence shows that CIITA is absolutely essential for MHC class II gene expression. In addition to regulating the transcription of class II genes, CIITA transfected into nonprofessional antigenpresenting cells (APCs) results in the expression of HLADM and invariant chain ［2］. CIITA is thus a key regulatory transcription factor to coordinate expression of genes required for HLA class IImediated immune responses［3］. We investigate the relationship between CIITA and HLADR expression in T cells, and explore its potential effect and significance. 毕业论文 Human peripheral blood mononuclear cells (PBMNC) were isolated from heparinized peripheral blood of normal donors using FicollHypaque densitygradient centrifugation and washed in Hanks’ balanced salt solution and used for experiments. To obtain a monocyteenriched cell population, PBMNC were incubated at 37℃ for 2 hours in plastic Petri dishes in RPMI 1640 culture medium (Gibco) containing 100 U/ml penicillin, 100 U/ml s