Dendritic Cell Immunotherapy for Cervical Cancer

描述(申请人提供)：人类乳头瘤病毒(HPV)16型和18型与宫颈癌的诱发和发展有关。本实验室的研究表明，HPV E7负载的树突状细胞(DC)是CD8+CTL的有效诱导者，具有杀伤宫颈癌患者自体肿瘤细胞的能力，DC还能够诱导抗原特异性的CD4+辅助T细胞反应。这些观察结果为HPVE7负载的DC免疫治疗宫颈癌提供了理论基础。核心假设是，HPV抗原负载的自体DC疫苗将在携带HPV16或HPV18基因的IB或IIA期宫颈癌患者中诱导抗原特异性和肿瘤特异性免疫反应。这一假设将在DC疫苗接种的第一阶段，即递增剂量临床试验中得到解决，共招募12名患者，覆盖3种剂量水平的DC疫苗。我们将使用来源于外周血单核细胞的完全成熟的自体DC，并负载HPVE7抗原的脂质体和锁孔帽状血蓝蛋白作为免疫示踪抗原。特定目标1将测试负载E7的DC疫苗是否能够在体内诱导延迟类型的E7特异性T细胞反应 E7抗原过敏皮肤试验。具体目标2将通过体外分析测试负载E7的DC疫苗是否能够诱导抗原特异性T细胞反应。实验室研究将包括HPV E7对CD4+T细胞和CD8+T细胞的增殖反应，以及ELISPOT对抗原特异性CD4+和CD8+T细胞细胞因子表达的评估。我们还将对识别已知的HPV16 E7表位的HLA*0201抗原限制性外周血CD8+T细胞进行HLA二聚体分析。这项研究将确定负载E7抗原的自体DC疫苗是否为HPV16或HPV18相关宫颈癌患者提供了潜在的治疗益处。

DESCRIPTION (provided by applicant): Human papillomavirus (HPV) types 16 and 18 are implicated in the induction and progression of cervical cancer. Studies from this laboratory have shown that HPV E7-loaded dendritic cells (DC) are effective inducers of CD8+ CTL with the ability to lyse autologous tumor cells from cervical cancer patients, and that DC are also able to induce antigen-specific CD4+ helper T cell responses. These observations provide a rationale for immunotherapy with HPV E7-loaded DC for the treatment of cervical cancer. The central hypothesis is that HPV antigen-loaded autologous DC vaccination will induce antigen-specific and tumor-specific immune responses in patients with stage IB or IIA cervical cancer carrying the HPV16 or HPV18 genotypes. This hypothesis will be addressed in a Phase I, escalating dose clinical trial of DC vaccination, enrolling a total of 12 patients and covering 3 dose levels of DC vaccination. We will use fully mature autologous DC derived from peripheral blood monocytes, and loaded by lipofection with HPV E7 antigen and keyhole limpet hemocyanin as an immunological tracking antigen. Specific Aim 1 will test whether E7 loaded DC vaccination is capable of inducing E7-specific T cell responses in vivo, as measured by delayed-type Hypersensitivity skin tests with E7 antigen. Specific Aim 2 will test whether E7-loaded DC vaccination is capable of inducing antigen-specific T cell responses as measured by in vitro analysis. Laboratory studies will include CD4+ T cell and CD8+ T cell proliferative responses to HPV E7, and assessment of cytokine expression by antigen-specific CD4+ and CD8+ T cells by ELISPOT. We will also conduct HLA dimer analysis of the frequencies of HLA* 0201 antigen-restricted peripheral blood CD8+ T cells recognizing known HLA* 0201-restricted HPV16 E7 peptide epitopes. This study will determine whether E7 antigen-loaded autologous DC vaccination offers the potential of therapeutic benefit for patients with HPV16 or HPV18-associated cervical cancer.