HSV-1多表位DNA疫苗眼表接种诱导粘膜免疫机制的研究

Herpes simplex keratitis (HSK) is the most common cause of corneal blindness in the world and is a chronic inflammatory process caused by the infection of herpes simplex virus type 1 (HSV-1). Development of a HSV-1 vaccine is a priority because these infections are common and cannot be well prevented. In the study, based on the immunology theory of epitope recognization and antigen presentation and by the method of bioinformatics, a novel HSV-1 multiepitope antigen molecule will be designed and DNA vaccine (pVAX1-MEG) mediated with liopsome will be constructed. The HSV-1 DNA vaccine will be prepared and immunized in the mice by ocular mucosal administration vs. intramuscular injection. The difference of immune responses and protection efficiency against HSV-1 challenge between ocular mucosal administration and intramuscular injection will be tested. Furhtermore, the change of lymphoid subpopulations and biological activity of ocular mucosal immune system (OMIS) will be compared with that of lacrimal gland, and mechanisms of ocular mucosal immunity induced by ocular mucosal delivery of HSV-1 multiepitode DNA vaccine will be explored. Collectively, the proposed studies will facilitate the development of more effective vaccines to inhibit or eliminate HSK in the future.

单纯疱疹病毒性角膜炎（Herpes Simplex Keratitis，HSK）居角膜病致盲病因的首位。本项目通过应用生物信息学分析技术，分析单纯疱疹病毒-1（Herpes Simplex Virus， HSV-1）gB、gD糖蛋白抗原表位特点，设计新型的病毒抗原表位串联基因（MEG）并构建重组真核表达质粒（pVAX1-MEG），利用阳离子脂质体作为载体，构建HSV-1 多表位DNA疫苗。比较肌肉注射和眼表粘膜接种两种接种途径所诱导的机体系统免疫和局部粘膜免疫免疫应答和免疫效果的差异，重点研究眼粘膜免疫系统（ocular mucosal immune system,OMIS）和泪腺中淋巴细胞亚群及其生物学功能的变化，探讨接种的HSV-1多表位基因疫苗所诱导的眼表粘膜免疫机制，为HSV-1 DNA疫苗的眼表粘膜接种提供理论依据，为HSK的防治提供新的疫苗接种策略。

单纯疱疹病毒性角膜炎（Herpes Simplex Keratitis，HSK）居角膜病致盲病因的首位。本研究通过应用生物信息学分析技术，分析单纯疱疹病毒-1（Herpes Simplex Virus， HSV-1）gB、gD糖蛋白抗原表位特点，设计新型的病毒抗原表位串联基因并构建HSV-1 多表位DNA疫苗，并比较肌肉注射和眼表粘膜接种两种接种途径所诱导的机体系统免疫和局部粘膜免疫免疫应答和免疫效果的差异。本研究成功构建HSV⁃1 多表位重组串联核酸疫苗；发现该疫苗无论是通过肌肉注射接种还是通过眼表粘膜免疫接种均可显著增强小鼠脾特异性CTL和NK细胞毒活性、脾T细胞增殖活性，血清IFN-γ、IL-4水平和血清特异性IgG滴度和中和抗体滴度，且两种接种方法间比较无显著性差异。肌注接种组和经眼表粘膜接种组泪液中IgA滴度和引流淋巴结培养液中均IFN-γ水平均增加，且经眼表粘膜接种组增加水平明显高于经肌肉接种组。病毒攻击实验结果也表明经眼表粘膜免疫接种预防HSV-1眼部感染效果优于肌肉接种。该研究为HSV-1 DNA疫苗的眼表粘膜接种提供理论依据，为HSK的防治提供新的疫苗接种策略。