人乳头瘤病毒11/6b型E7蛋白对浆细胞样树突状细胞中TLR/MyD88信号传导通路的影响

Persistence and recurrence is the major difficulty of condyloma accuminatum (CA) treatment, because effective cellular immunity couldn't be induced to clear the HPV infection with decreased immune function on the lesions. Plasmacytoid dendritic cells (pDC) as an important subsets of dendritic cells, play an important role of connecting the natural immune response and the adaptive immune response in the immune system. PDCs can secrete type I interferon with antiviral activity, and also cause persistent infection due to virus infection-induced immune tolerance.Our previous study found pDC infiltration in the interstitial condyloma lesions, and HPV 11 E7 CTL epitope peptides combined with TLR agonists could significantly promote the differentiation,maturation and functional activity of monocyte-derived dendritic cells (mdDC) and pDC and, and further enhanced mdDC induced specific antiviral cellular immune response. Based on this, this project intends to use the condyloma acuminata associated HPV type 11 and 6b major pathogenic gene E7 proteins, combined with TLR7 and TLR9 agonists and antagonists, stimulating human peripheral blood separated pDCs in vitro, to study the influence on TLR/MyD88 signaling pathway in pDCs, in order to explore the possible mechanism of pDC-induced immune responses, which will be conducive to further establish experimental and theoretical basis for the study on the immunotherapy targeting the regulation of pDCs function.

尖锐湿疣的迁延与复发是治疗的难点，原因在于皮损局部免疫功能下调，机体无法诱导有效的细胞免疫清除HPV感染。浆细胞样树突状细胞(pDC)可分泌具抗病毒作用的I型干扰素，亦可因病毒感染诱导免疫耐受而导致持续感染, 在机体免疫系统中具有连接天然免疫反应和适应性免疫反应的作用。我们前期研究发现尖锐湿疣皮损间质中有pDC浸润，HPV 11 E7 CTL表位多肽联合TLR激动剂可明显促进单核细胞来源树突状细胞(mdDC)和pDC的分化成熟和功能活性，并进一步增强mdDC诱导的特异性抗病毒细胞免疫应答。在此基础上，本项目拟采用主要引起尖锐湿疣感染的HPV11和6b型的主要致病基因E7蛋白及TLR7和TLR9的激动剂及抑制剂，体外刺激人外周血分离的pDCs，研究其对pDC中TLR/MyD88信号传导通路的影响，探索其可能诱导的免疫应答机制，为研究基于靶向调控pDC功能的免疫治疗手段提供一定的实验和理论基础

尖锐湿疣的迁延与复发是治疗的难点，原因在于皮损局部免疫功能下调，机体无法诱导有效的细胞免疫清除人乳头瘤病毒（HPV）感染。浆细胞样树突状细胞(pDC)可分泌具抗病毒作用的I型干扰素，亦可因病毒感染诱导免疫耐受而导致持续感染, 在机体免疫系统中具有连接天然免疫反应和适应性免疫反应的作用。我们前期研究发现尖锐湿疣皮损间质中有pDC浸润，HPV 11 E7 CTL表位多肽联合TOLL样受体（TLR）激动剂可明显促进单核细胞来源树突状细胞(mdDC)和pDC的分化成熟和功能活性，并进一步增强mdDC诱导的特异性抗病毒细胞免疫应答。在此基础上，本项目拟采用主要引起尖锐湿疣感染的HPV11和6b型的主要致病基因E7蛋白联合TLR7和TLR9的激动剂，体外刺激人外周血分离的pDCs，研究其对pDC中TLR/MyD88信号传导通路的影响，探索其可能诱导的免疫应答机制。研究结果发现，HPV11/6b E7蛋白可诱导pDC的分化成熟，增强其抗原递呈功能，激活TLR7和TLR9通路而对pDC的功能产生影响，诱导I型干扰素和促炎症因子IL-6的表达，且在TLR7/9激动剂的刺激下更为显著。但对Myd88/TLR通路的相关蛋白的表达影响不大。证明低危型HPV 6b/11 E7蛋白较高危型HPV16E7蛋白具有更强的免疫原性，在感染初期可激活天然免疫，增强pDC免疫功能，而TLR7激动剂咪喹莫特可显著增强其免疫原性，初步阐明了临床咪喹莫特治疗尖锐湿疣的可能机制，为进一步研究基于靶向调控pDC功能的免疫治疗药物提供理论和实验基础。..