CYTOTOXIC T CELL RESPONSES TO HPV 16 EARLY GENE PROTEINS

细胞毒性 T 细胞对 HPV 16 早期基因蛋白的反应

• 批准号: 2098696
• 负责人: Dennis J. McCance
• 金额: $ 19.08万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1996-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2098696
• 关键词: athymic mouse; cell mediated cytotoxicity; cell mediated lymphocytolysis test; cellular immunity; chimeric proteins; cytotoxic T lymphocyte; epitope mapping; genetically modified animals; human papillomavirus; immunization; laboratory mouse; recombinant proteins; regulatory gene; spleen; tissue /cell culture; transfection; virus antigen; virus protein

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.

人乳头瘤病毒（HPV）16是最常见的病毒 宫颈癌。 对细胞介导的响应知之甚少 HPV 16抗原，尽管初步数据表明肿瘤保护 对E6和E7蛋白的反应是引起的。 我们以前绘制过 E7的T-助细胞表位，现在建议检查特异性 细胞毒性T淋巴细胞（CTL）对HPV 16的响应的保护价值和保护价值 表位。 最初，我们将研究CTL对E6和E7的响应 体外和体内蛋白质。 但是，由于HPV 16不能 在体外或实验动物中传播，有必要使用 新方法。 因此，我们建议： 1。研究CTL反应首先在体外对E6和E7蛋白的研究 两者都启动： 一个。用表达E6或 E7，或 b。含有全长E6或E7的离子病毒重组剂 蛋白质。 该矢量允许表达整个重组 蛋白质。 2。确定CTL响应是否针对HPV 16肿瘤有保护 产生细胞，如果相同的响应会导致回归 现有肿瘤。 3。绘制使用T细胞中保护蛋白中包含的表位 克隆。 4。研究这些肽是否掺入主要抗原 Sabin的部位减弱1型脊髓灰质炎病毒将刺激保护性 小鼠中的CTL。 由于小鼠通常不接受脊髓灰质炎病毒 复制，含有人脊髓灰质炎病毒受体的转基因小鼠和 将使用允许病毒复制。 免疫的脾细胞 转基因将在体外测试特定的HPV 16 CTL和 重建的裸鼠的肿瘤保护和回归。 结果将阐明这些向量的有用性 递送肽以产生保护性粘膜和系统性 免疫。 此外，这些嵌合体可能是潜在HPV的基础 疫苗，具有多种HPV类型的表现的优势 可以合并。