CYTOTOXIC T CELL RESPONSES TO HPV 16 EARLY GENE PROTEINS

人乳头瘤病毒（HPV）16型是最常见的病毒相关 子宫颈癌 对细胞介导的对 HPV 16抗原，尽管初步数据表明， 对E6和E7蛋白质引起应答。 我们之前绘制了 E7的辅助性T细胞表位，现在建议检查特异性 细胞毒性T淋巴细胞（CTL）对HPV16的保护作用 表位 首先，我们将研究CTL对E6和E7的反应， 蛋白质在体外和体内。 然而，由于HPV 16不能被 在体外或实验室动物中繁殖，将需要使用 新方法。 因此，我们建议： 1.首次在体外研究小鼠对E6和E7蛋白的CTL应答 用以下两种物质之一预处理： a.用表达E6的质粒构建体转染的同基因细胞或 E7或 B.含有全长E6或E7的重组痘苗病毒 proteins. 该载体允许表达整个重组 蛋白 2.确定CTL应答是否对HPV 16肿瘤具有保护作用 生产细胞，如果相同的反应将导致退化 现有肿瘤 3.使用T细胞定位保护性蛋白内所含的表位 克隆 4.研究这些肽是否掺入主要抗原 Sabin减毒脊髓灰质炎病毒1型的位点将刺激保护性 小鼠中的CTL。 由于小鼠通常不允许脊髓灰质炎病毒 复制，含有人脊髓灰质炎病毒受体的转基因小鼠， 将使用允许病毒复制的方法。 免疫小鼠脾细胞 转基因将在体外测试特异性HPV 16 CTL， 在重组裸鼠中的肿瘤保护和消退。 结果将阐明这些载体作为一种手段的有用性 提供肽来产生保护性粘膜和全身 免疫力 此外，这些嵌合体可能是潜在HPV的基础。 疫苗，其优点是来自多于一种HPV类型的表位 可以合并。

Human papillomavirus (HPV) type 16 is the most common virus associated with cervical cancer. Little is known of the cell mediated responses to HPV 16 antigens, although preliminary data suggests that tumor protection responses are elicited to E6 and E7 proteins. We have previously mapped T-helper cell epitopes of E7 and now propose to examine the specificity and protective value of cytotoxic T-lymphocyte (CTL) responses to HPV 16 epitopes. Initially, we will investigate CTL responses to E6 and E7 proteins in vitro and in vivo. However, since HPV 16 cannot be propagated in vitro or in laboratory animals it will be necessary to use new approaches. We, therefore, propose to: 1. Study CTL responses first in vitro to E6 and E7 proteins in mice primed with either: a. syngeneic cells transfected with plasmid constructs expressing E6 or E7, or b. vaccinia virus recombinants containing the full length E6 or E7 proteins. This vector allows expression of the whole recombinant protein. 2. Determine whether CTL responses are protective against HPV 16 tumor producing cells and if the same responses will cause regression of existing tumors. 3. Map the epitopes contained within protective proteins using T cell clones. 4. Investigate if these peptides incorporated into the major antigenic site of the Sabin attenuated poliovirus type 1 will stimulate protective CTLs in mice. Since mice are normally non-permissive for poliovirus replication, transgenic mice containing the human poliovirus receptor and allowing viral replication will be used. Spleen cells from immunized transgenics will be tested for specific HPV 16 CTLs in vitro and for tumor protection and regression in reconstituted nude mice. The results will shed light on the usefulness of these vectors as a means of delivering peptides to produce protective mucosal and systemic immunity. Moreover, these chimeras may be the basis for a potential HPV vaccine, with the advantage that epitopes from more than one HPV type could be incorporated.