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RECOGNITION OF LEUKEMIA ANTIGEN BY ANTI-VIRAL CTL'S

抗病毒 CTL 对白血病抗原的识别

基本信息

批准号：
3458236
负责人：
HILLARY D. WHITE
金额：
$ 9.75万
依托单位：
DARTMOUTH COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
1987
资助国家：
美国
起止时间：
1987-07-01 至 1993-06-30
项目状态：
已结题

项目摘要

We propose to identify ways in which anti-viral CTL recognize or fail to recognize leukemia virus cell surface antigen in the murine system. In particular, we plan to study why normal cells from AKXL mice, which carry the Akv-1 murine leukemia provirus, are recognized by C57B1/6 anti-AKR/Gross virus CTL, whereas cells from other AKXL mice which carry the highly related Akv-4 provirus, are not recognized by such antiviral CTL's. Green and coworkers have used standard immunological techniques including CTL recognition and induction assays and antiviral monoclonal antibody recognition assays that clearly indicate the importance of the viral antigen gp70 in the CTL recognition process. To further study viral epitope expression for recognition by CTL, I propose here to clone and compare by restriction mapping the Akv-1 and Akv-4 proviral DNA loci. The cell surface viral antigens expressed by these proviruses and recognized by CTL will then be studied and compared by transfecting recombinant proviral subclones into fibroblast or lymphoblastoid cell lines. After culturing the transfected cells in G418 selective medium, viral antigen expression will be assayed for by cytofluorography using antiviral monoclonal antibodies and by CTL functional assays. By these means, we can better define the viral elements necessary for CTL recognition and induction and better understand the promotion of tumor establishment resulting from the failure of these processes.

我们建议确定抗病毒CTL识别或不能识别小鼠白血病病毒细胞表面抗原系统特别是，我们计划研究为什么正常细胞从携带Akv-1鼠白血病原病毒的AKXL小鼠，被C57 B1/6抗AKR/Gross病毒CTL识别，而细胞来自其他携带高度相关Akv-4的AKXL小鼠前病毒不能被这种抗病毒CTL识别。绿色和同事们使用标准的免疫技术， CTL识别和诱导测定以及抗病毒单克隆抗体抗体识别试验清楚地表明了病毒抗原gp 70的CTL识别过程。到进一步研究CTL识别的病毒表位表达，在此建议通过限制性酶切图谱克隆和比较 Akv-1和Akv-4前病毒DNA基因座。细胞表面病毒由这些前病毒表达并被CTL识别的抗原将然后通过克隆重组体进行研究和比较将前病毒亚克隆入成纤维细胞或淋巴母细胞样细胞系。在G418选择性培养基中培养转染的细胞后，将通过细胞荧光照相术测定病毒抗原表达使用抗病毒单克隆抗体和通过CTL功能测定。通过这些方法，我们可以更好地定义病毒元素 CTL识别和诱导所必需的，并且更好地了解促进肿瘤形成的原因，这些过程的失败。