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

TONG Xiao; SHEN Tao; LIANG Hua; MENG Qing-lai; ZHONG Wei-zhou; MA Yan; WEI Li-li; XIANG Wen-hua; SHEN Rong-xian; XU Jian-qing; ZHANG Xiao-yan; SHAO Yi-ming

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December 2005

TONG Xiao, SHEN Tao, LIANG Hua, MENG Qing-lai, ZHONG Wei-zhou, MA Yan, WEI Li-li, XIANG Wen-hua, SHEN Rong-xian, XU Jian-qing, ZHANG Xiao-yan and SHAO Yi-ming. A Novel Flow Cytometric Assay for Evaluating Antigen-specific CTL Responses and Its Application in EIAV Immunological Study[J]. Virologica Sinica, 2005, 20(6): 642-646.

Citation: TONG Xiao, SHEN Tao, LIANG Hua, MENG Qing-lai, ZHONG Wei-zhou, MA Yan, WEI Li-li, XIANG Wen-hua, SHEN Rong-xian, XU Jian-qing, ZHANG Xiao-yan, SHAO Yi-ming. A Novel Flow Cytometric Assay for Evaluating Antigen-specific CTL Responses and Its Application in EIAV Immunological Study .VIROLOGICA SINICA, 2005, 20(6) : 642-646.

利用流式细胞术测定CTL活性方法在EIAV免疫学中的应用

童骁 ^1,2 ,
沈弢 ² ,
梁华 ² ,
孟庆来 ^1,2 ,
钟卫洲 ¹ ,
马燕 ² ,
魏丽丽 ³ ,
相文华 ³ ,
沈荣显 ³ ,
徐建青 ² ,
张晓燕 ^2* ,
邵一鸣 ²

1.
1.湖北武汉中国科学院武汉病毒研究所分子病毒学重点实验室 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 3.中国农业科学院哈尔滨兽医研究所黑龙江哈尔滨

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。
- 细胞毒性T淋巴细胞（CTL）
- , 马传染性贫血症病毒（Equine infectious anemia virus EIAV）
- , 流式细胞测定术

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

Abstract

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.
- CTL
- , EIAV
- , Flow Cytometry

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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

Abstract: 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.