Multiple Antigen-Engineered DC Immunization and IFNalpha-2b Boost for Metastatic Melanoma

本项目试验了一种改进的DC疫苗，旨在促进体内交叉呈现和决定性传播。基于我们之前的试验结果，我们已经做了几个重要的改进：用3个全长的、明确的肿瘤抗原来设计DC，以激活多个CD8+和CD4+ T细胞克隆（减少对抗原丢失变异的担忧）；为DC的生命周期提供抗原呈递；向CD8+ T细胞提供同源的CD4+ T细胞帮助（“帮助”CTL）；使用成熟的DC（一种与腺病毒（AdV）转导信号特异性匹配的鸡尾酒）；通过NK细胞迁移和活化激活先天免疫；系统性IFNα增强（内源性DC 1型倾斜，改善交叉启动和对T细胞的直接作用）。总之，这种疫苗策略应该更有效地激活包含多种适应性和先天效应的多克隆抗肿瘤反应，我们预测这将导致更高频率的患者不仅激活疫苗编码的抗原特异性T细胞反应，而且还会产生决定因素

This project tests an improved DC vaccine which is designed to promote in vivo cross presentation and determinant spreading. Based on results from our previous trials, we have made several important improvements: engineering the DC with 3 full-length, defined, tumor antigens to activate multiple CD8+ and CD4+ T cell clones (reducing the concern for antigen loss variants); providing antigen presentation for the life of the DC; providing cognate CD4+ T cell help to the CD8+ T cells ("helped" CTL); using a matured DC (a cocktail specifically matched to adenovirus (AdV) transduction signals); activating innate immunity via NK cell migration and activation; and boosting with systemic IFNα (for endogenous DC type 1 skewing, improved cross-priming and direct effects on T cells). Together, this vaccine strategy should more potently activate a polyclonal anti-tumor response incorporating multiple adaptive and innate effectors which we predict will lead to a higher frequency of patients who not only activate vaccine-encoded antigen-specific T cell responses, but also develop determinant spreading and a significant clinical response. The three aims are: Aim 1: Completion of the AdVTMM2/DC +/- IFNα clinical trial 1.a. Clinical outcomes in the stage IV patients; 1.b. Immunologic outcomes (from blood and TIL; baseline, post-vaccine and post-IFNα); 1.c. AdV-specific cellular and humoral responses. Aim 2: Mechanism and biomarkers of clinical response: 2.a. DC Vaccine Transcriptional Profiling (+/- maturation, +/- AdVTMM2); 2.b. Tumor Transcriptional Profiling (baseline, post-vaccine and post-IFNα); 2.c. Peripheral blood signaling induced by IFNα (STATs); 2.d. Serum profiling (autoimmunity antibodies, LDH, CRP cytokines); 2.e. Molecular mimicry with mycoplasma (impact of baseline memory to mycoplasma); 2.f. Germline DNA SNP analysis Aim 3: Mechanism of NK cell "activation" in anti-melanoma immunity In vitro studies to expand our recent findings in AdV/DC-NK cell cross-talk with banked melanoma patient blood and tumor (primary tumor expanded to cell lines): 3.a. Direct NK cell interactions with melanoma tumor cells (cytotoxicity, cytokines); 3b: Helper/type 1 skewing role in shaping adaptive CD8+ and CD4+ T cell responses; 3c: Melanoma tumor impact on NK cell function.