CYTOTOXIC T CELL RESPONSES TO HPV 16 EARLY GENE PROTEINS

人乳头瘤病毒(HPV)16型是最常见的相关病毒 患有宫颈癌。对细胞介导的反应知之甚少。 HPV16抗原，尽管初步数据表明，肿瘤保护 对E6和E7蛋白有反应。我们之前已经绘制了 E7的T辅助细胞表位，现在建议检测其特异性 细胞毒性T淋巴细胞(CTL)对HPV16免疫应答的保护作用 表位。首先，我们将调查对E6和E7的CTL反应 体外和体内的蛋白质。然而，由于HPV16不能 在体外繁殖或在实验动物中繁殖将有必要使用 新方法。因此，我们建议： 1.首次研究小鼠CTL对E6和E7蛋白的体外应答 以以下任一为基准点： A.转染表达E6或E6的同源细胞 E7，或 B.含有全长E6或E7的痘苗病毒重组体 蛋白质。该载体可表达完整的重组蛋白 蛋白。 2.确定CTL反应是否对HPV16肿瘤具有保护作用 产生细胞，如果同样的反应会导致 现有的肿瘤。 3.使用T细胞绘制保护性蛋白中包含的表位 克隆人。 4.研究这些多肽是否与主要抗原结合 沙宾减毒1型脊髓灰质炎病毒的位置将刺激保护性 小鼠体内的CTL。由于小鼠通常不允许携带脊髓灰质炎病毒 复制，含有人脊髓灰质炎病毒受体和 允许病毒复制将被使用。免疫后的脾细胞 转基因药物将在体外测试特定的HPV16CTL，并检测 重组裸鼠体内肿瘤的保护和消退。 结果将阐明这些向量作为一种手段的有用性 提供多肽以产生保护性的粘膜和全身 豁免权。此外，这些嵌合体可能是潜在的HPV的基础。 疫苗，其优点是表位来自一种以上的HPV类型 可能会被合并。

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.