HUMAN PAPILLOMAVIRUS SPECIFIC T-CELL RESPONSES

我们的长期目标是识别人乳头瘤病毒(HPV)人工合成 有可能作为免疫治疗和/或疫苗试剂的多肽 抗HPV相关的宫颈肿瘤。我们和其他人已经证明了 HPV与大多数浸润性癌和浸润性癌有关 (宫颈上皮内瘤变)，并已被发现在 人类生殖器癌发生的重要途径。奇怪的是，有些人 HPV感染的患者会患上宫颈肿瘤，而其他人则不会。 此外，细胞免疫缺陷的患者也增加了 人乳头瘤病毒相关疾病的流行。因此，重要的是， 了解细胞介导的免疫反应(辅助性T细胞和细胞毒性T细胞 细胞免疫)，以便制定有效的策略 HPV阳性肿瘤的免疫预防和免疫治疗。我们建议 HPV相关性宫颈上皮内瘤变患者HPV特异性T细胞反应的检测 并将它们的水平与没有HPV迹象的女性进行比较 感染和/或CIN。我们还将把这些T细胞反应与 那些已经接受治疗并仍处于疾病中的女性- 无症状的或有复发或持续性疾病的。这将是 通过以下步骤完成：(A)制备从HPV衍生的合成肽 编码E6/E7癌蛋白及其与重组E6、E7的联合应用 患者白细胞T细胞增殖研究中的蛋白质，(B) 确定这些多肽是否使人类白细胞抗原相合的靶细胞增敏 由患者的细胞毒性T淋巴细胞(CTL)裂解，以及(C)执行 血液和组织活检标本的双色免疫荧光分析 对于HPV特异性的T4和T8细胞。而主要的焦点将是多肽 从肿瘤中最常见的HPV蛋白E6和E7来看，我们 还将在文献中搜索来自其他HPV的其他多肽 蛋白质。这些研究的阳性结果将识别HPV多肽 是肿瘤特异性T细胞抗原。这些研究将提供 为未来对高危患者进行免疫治疗干预奠定基础 宫颈癌。

Our long-term goal is to identify Human Papillomavirus (HPV) synthetic peptides that have potential as immunotherapeutic and/or vaccine reagents against HPV-associated cervical neoplasia. We and others have shown that HPV is associated with the majority of preinvasive and invasive carcinomas (CIN) of the uterine cervix and has been found to contribute in a significant way to the genesis of human genital cancer. Curiously, some HPV infected patients develop cervical neoplasms while others do not. Also, patients with defects in cell mediated immunity have an increased prevalence of HPV-associated diseases. It is, therefore, important to understand cell mediated immune responses (both helper T- and cytotoxic T- cell immunity) in order to develop strategies for efficient immunoprevention and immunotherapy of HPV-positive tumors. We propose to determine HPV-specific T cell responses in women with HPV-associated CIN and compare their levels with those in women with no signs of HPV infection and/or CIN. We will also compare these T cell responses to those in women that have undergone treatment and remained in a disease- free condition or with recurrent or persistent disease. This will be accomplished by: (a) preparing synthetic peptides derived from HPV encoded E6/E7 oncoproteins and using them along with recombinant E6 and E7 proteins in T-cell proliferation studies on patient white blood cells, (b) determining whether these peptides sensitize HLA-matched target cells for lysis by cytotoxic T lymphocytes (CTLs) from patients, and (c) performing two-color immunofluorescence analysis of blood and tissue biopsy samples for HPV-specific T4- and T8-cells. While the major focus will be peptides from E6 and E7, the HPV proteins most commonly expressed in tumors, we will also search the literature for additional peptides from other HPV proteins. Positive results from these studies would identify HPV peptides that are tumor-specific T-cell antigens. These studies will provide the basis for future immunotherapeutic intervention in patients at risk for cervical cancer.