Ⅱ类抗原反式激活因子和人白细胞抗原-DR检测方法的比较  

Comparesion of the method of CⅡTA and HLA-DR detection

作　　者：李慧[1] 徐开林[1] 潘秀英[1] 李振宇[1] 鹿群先[1] 

机构地区：[1]徐州医学院附属医院血液科,徐州221002

出　　处：《中华检验医学杂志》2003年第8期463-465,共3页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目 ( 30 1 70 389)

摘　　要：目的　研究Ⅱ类抗原反式激活因子 (CⅡTA)与人白细胞抗原 DR(HLA DR)表达的相关性 ,探讨CⅡTA在移植物抗宿主病 (GVHD)中检测的意义。方法　从健康人外周血分离获得T细胞 ,给予干扰素 γ(IFN γ) 10 0 0U /ml后 ,在不同时间用逆转录聚合酶链反应 (RT PCR )检测CⅡTAmRNA表达 ,用免疫印迹法检测HLA DR抗原的表达。然后 ,给予Stat1α反义寡核苷酸 (AS)抑制CⅡTA ,分别检测CⅡTAmRNA及HLA DR抗原在各自表达量最高时间点的变化。结果　CⅡTAmRNA在IFN γ作用后 5h开始表达 ,至 14h表达量最高 ,此后逐渐下降 ;HLA DR抗原在 2 8h可检测出 ,52h表达量最高 ,此后逐渐下降 ,至 76h仅可检测到少量抗原的表达 ;给予AS后与对照组相比 ,CⅡTAmRNA和HLA DR表达量均下降。结论　CⅡTA与HLA DR表达成正相关性 ,且先于HLA DR出现 。Objective To investigate the relation and difference of expression phase between classⅡtransactivator (CⅡTA) and HLA-DR antigens after IFN-γ incubation, so as to investigate the potential effect of the class Ⅱ trasactivator (CⅡTA) in graft-versus-host disease(GVHD). Methods T lymphocyte from peripheral blood of health was incubated with IFN-γ for 1 000 U/ml. RT-PCR was used to detect CⅡTA mRNA and Western blot was used to explore HLA-DR antigen in various time periods. Then Stat1α antisense oligonucleotides (AS) were given to inhibit the expression of CⅡTA, CⅡTA mRNA and HLA-DR antigen were tested again at peak point. Results CⅡTA mRNA was detectable 5 h after IFN-γ treatment and peaks at 14 h; HLA-DR protein was detectable 28 h after IFN-γ treatment and peaks at 52 h. The expression of CⅡTA mRNA and HLA-DR protein was lower in AS groups than that in control groups. Conclusion CⅡTA expression was positively correlated to HLA-DR expression, and earlier than the HLA-DR expression after IFN-γ incubation. CⅡTA might be used as an early predicting marker of GVHD.

关 键 词：Ⅱ类抗原反式激活因子 人白细胞抗原-DR 检测方法 移植物抗宿主病 CⅡTA 逆转录聚合酶链反应 

分 类 号：R446.6[医药卫生—诊断学]