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

该项目测试了一种改进的DC疫苗，该疫苗旨在促进体内交叉呈现和决定簇传播。根据我们以前的试验结果，我们做了几项重要的改进：用3个全长、明确的肿瘤抗原工程DC来激活多个CD8+和CD4+T细胞克隆(减少了对抗原丢失变体的担忧)；为DC的生命提供抗原递呈；为CD8+T细胞提供同源的CD4+T细胞帮助(帮助的CTL)；使用成熟的DC(与腺病毒(Adv)信号转导信号特异性匹配的鸡尾酒)；通过NK细胞的迁移和激活激活固有免疫；以及使用全身干扰素α增强(针对内源性DC 1型偏斜，改进的交叉免疫和对T细胞的直接作用)。总而言之，这种疫苗策略应该更有效地激活包含多个适应性和先天效应器的多克隆抗肿瘤反应，我们预测这将导致患者不仅激活疫苗编码的抗原特异性T细胞反应的频率更高，而且还会产生决定因素 扩散和显著的临床反应。 三个目标是：目标1：完成AdVTMM2/DC+/-干扰素α临床试验1.A.IV期患者的临床结局；1.免疫学结果(来自血液和TIL；基线、疫苗后和干扰素后α)；ADV特异性的细胞和体液反应。 目的2：临床反应的机制和生物标志物：2.a。DC疫苗转录图谱(+/-成熟，+/-AdVTMM2)；2.b.肿瘤转录图谱(基线、疫苗后和干扰素α后)；干扰素α(STATS)诱导外周血信号传导；血清学分析(自身免疫抗体、LDH、CRP细胞因子)；支原体分子拟态(基线记忆对支原体的影响)；生殖系DNA SNP分析 目的3：NK细胞活化在抗黑色素瘤免疫中的作用机制 体外研究扩展了我们最近在ADV/DC-NK细胞与储存的黑色素瘤患者血液和肿瘤(原发肿瘤扩大到细胞系)之间的相互作用：3.a.NK细胞与黑色素瘤细胞的直接相互作用(细胞毒性、细胞因子)；3b：辅助者/1型偏斜在形成适应性CD8+和CD4+T细胞反应中的作用；3c：黑色素瘤肿瘤对NK细胞功能的影响。

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.