利用流式细胞术测定CTL活性方法在EIAV免疫学中的应用  

A Novel Flow Cytometric Assay for Evaluating Antigen-specific CTL Responses and Its Application in EIAV Immunological Study

作　　者：童骁[1] 沈弢 梁华 孟庆来[1] 钟卫洲[1] 马燕 魏丽丽[3] 相文华[3] 沈荣显[3] 徐建青 张晓燕 邵一鸣 

机构地区：[1]中国科学院武汉病毒研究所分子病毒学重点实验室,湖北武汉430071 [2]中国疾病预防与控制中心国家性病艾滋病预防控制中心,北京100050 [3]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001

出　　处：《中国病毒学》2005年第6期642-646,共5页Virologica Sinica

基　　金：国家自然科学基金资助(30371319)。

摘　　要：利用PKH-26和CFSE两种荧光染料对靶细胞染色,建立了一种通过流式细胞术进行马传染性贫血症病毒 (Equine infectious anemia virus,EIAV)抗原特异性细胞毒性T淋巴细胞(Cytotoxic T lymphocytes,CTL)反应的 新方法,避免了经典的Cr51释放法对检测人员的放射线威胁,降低了本底释放,提高了检测的灵敏度。将该检测方 法用于检测EIAV疫苗毒接种马和嵌合克隆接种马的细胞免疫反应变化趋势,数据显示细胞免疫反应在接种后3 个月达到成熟阶段而后保持在较高的反应水平。该方法的成功建立和应用为研究EIAV减毒疫苗的免疫机制提 供了好的研究手段,也为其他病毒的免疫学研究提供了新的参考方法。A novel flow cytometric assay for evaluating Equine infectious anemia virus （EIAV） antigen-specific CTL responses was established and by using PKH-26 and CFSE as cell colouration dyes. Compared with the traditional 51Cr -release assay, the new method is more sensitive, has a decrease background and avoids the risk of radioactive exposure to workers. The cellular immuno-response against EIAV infection was detected using this method. The maturation of EIAV antigen-specific CTL response was characterized. The result showed that the CTL response reached a maximum at three months and remained relatively high. The novel method is directly applicable to research on the immune mechanism of the EIAV attenuated vaccine.

关 键 词：细胞毒性T淋巴细胞(CTL) 马传染性贫血症病毒(Equine in fectious ANEMIA virus EIAV) 流式细胞测定术 

分 类 号：S852.65[农业科学—基础兽医学]