抗细胞因子主动免疫调控肿瘤微环境及其协同治疗性HPV疫苗的抗肿瘤免疫效应与机制

Persistent infection of human papillomavirus（HPV）is the pathogenic cause of human cervical cancer and also some other malignant tumors. Present studies on therapeutic HPV vaccine mostly focus on inducing the production of tumor antigen-specific effector immune cells. However, there were no clinical trials of therapeutic HPV vaccine which aimed to treat cervical intraepithelial neoplasia (CIN) or cervical cancer have presented significant clinical efficacy. The reason is most probably due to immune suppression of tumor microenvironment. The currently proposed project will employ a new strategy of anticytokine active immunization against immune suppressive factors such as IL-4，IL-13，TGF-β1, IL-10 and VEGF to induce specific antibodies and modulate tumor microenvironment on a persistent basis; further, the combination of multiple cytokine targets or with an antigen-specific therapeutic vaccination will be performed; and, the studies will investigate the differentiation, proliferation, activation, and infiltration into tumor tissue of immune suppressive cells, including regulatory T dells (Tregs), tumor-associated macrophage (TAM) and myeloid derived suppressor cells (MDSCs), and of tumor-killing effector cells including cytotoxic T lymphocytes (CTLs) and Th1 cells, as well as their actual function statuses such as the expression of functional mediators and activation of signal pathways. The studies will demonstrate the changes and features of tumor microenvironment and anti-tumor immune which are under the persistent intervening of anti-cytokine immune, and discover the involved molecular and cellular mechanisms. In summary, the proposed project sought to further our knowledge of tumor immune suppression and escape, and develop a new and promising vaccine strategy by combining modulating tumor microenvironment and promoting antigen -specific cellular immunity.

HPV感染引起的宫颈癌等疾病危害严重，其治疗性疫苗在临床研究中尚缺乏疗效。本项目旨在探讨通过调控肿瘤免疫抑制微环境，促进抗肿瘤效应细胞的产生、瘤组织聚集及抗瘤活性保持，以增强疫苗临床应用潜力。项目创新性地以抗细胞因子主动免疫诱导特异抗体产生，持续性拮抗宫颈癌前病变或癌组织中高度表达、并在免疫抑制微环境产生与发展中可能起关键作用的细胞因子如IL-4、IL-13、TGF-β1、IL-10与VEGF等，探讨其单独、多靶点组合及与肿瘤抗原特异治疗性HPV疫苗联合应用的抗肿瘤效应，明确有潜力的细胞因子靶点及联合策略；同时，考察免疫抑制细胞Treg、TAM、MDSC以及效应细胞CTLs、Th1等的增殖、激活、肿瘤组织浸润以及功能状态（如信号通路激活与功能介质表达等），了解免疫干预产生的肿瘤微环境与抗肿瘤免疫应答的改变及特点，揭示关键效应机制。本项目将为宫颈癌等肿瘤疾病的治疗性疫苗免疫干预提供新的策略。

人乳头瘤病毒（HPV）感染可引起宫颈癌等生殖道癌以及头颈癌等非生殖道癌，严重危害人类健康。治疗性疫苗是有潜力的肿瘤免疫治疗策略，但在目前临床研究中尚缺乏显著疗效。分析其主要原因，可能由于疫苗激发肿瘤特异的细胞免疫应答的能力不足，以及肿瘤形成的免疫抑制、免疫逃逸机制尤其是免疫抑制的肿瘤微环境限制了效应细胞的产生及作用发挥。本项目旨在探讨调控肿瘤免疫抑制微环境，促进抗肿瘤效应细胞的产生、瘤组织聚集及抗瘤活性保持，以增强疫苗临床应用潜力。项目创新性地以抗细胞因子主动免疫诱导特异抗体产生，以持续性拮抗宫颈癌前病变或癌组织中高度表达、并在免疫抑制微环境产生与发展中可能起关键作用的细胞因子如IL-4、IL-13、IL-33、TGF-β1、FGF-2与VEGF等，探讨其单独、多靶点组合对肿瘤免疫的影响及其与肿瘤抗原特异治疗性HPV疫苗联合应用的抗肿瘤效应，明确有潜力的细胞因子靶点及疫苗联合干预策略；此外，本项目研究还致力于重组细菌囊泡、自折叠肽介导的肽纳米纤维、佐剂修饰的PLGA颗粒等新型纳米肿瘤疫苗的研究。结果显示，靶向2型免疫关键介质（IL-4、IL-13、IL-33）及重要促血管生成因子（FGF-2与VEGF）的主动免疫，可有效的下调荷瘤小鼠免疫抑制性细胞Treg、MDSC及Th2的产生及肿瘤浸润，而有效刺激效应细胞CTLs与Th1的增殖、激活、肿瘤组织浸润以及功能状态保持；同样，新型的纳米疫苗有效激发了抗肿瘤细胞免疫应答；通过抗细胞因子免疫调控及纳米疫苗免疫的联合干预，在实验小鼠中产生了较之单一手段更为强劲的抗肿瘤免疫，更显著的抑制了肿瘤的生长及转移。本项目通过了解免疫干预产生的肿瘤微环境与抗肿瘤免疫应答的改变及特点，揭示了关键效应机制，为研发宫颈癌等肿瘤疾病的新型治疗性疫苗及免疫干预策略、取得临床疗效突破提供了重要基础。