结核分枝杆菌EsxV脂质体纳米颗粒亚单位疫苗的制备及免疫学特性  

Preparation of Mycobacterium tuberculosis EsxV lipid nanoparticles subunit vaccine and its immunological characteristics

作　　者：白鹭 路延之 宁唤唤 康亚莉 谢燕玲 康健 李雪 崔若囡 韦垠 刘月芹[2] 柏银兰 BAI Lu;LU Yanzhi;NING Huanhuan;KANG Yali;XIE Yanling;KANG Jian;LI Xue;CUI Ruonan;WEI Yin;LIU Yueqin;BAI Yinlan(Department of Microbiology and Pathogen Biology,Basic Medical School,Air Force Medical University,Xi’an 710032,Shaanxi,China;School of Life Sciences,Yan’an University,Yan’an 716000,Shaanxi,China;College of Life Sciences,Northwest University,Xi’an 710069,Shaanxi,China;Department of Physiology and Neurobiology,Basic Medical School,Ningxia Medical University,Yinchuan 750001,Ningxia,China)

机构地区：[1]空军军医大学基础医学院微生物与病原生物学教研室,陕西西安710032 [2]延安大学生命科学学院,陕西延安716000 [3]西北大学生命科学学院,陕西西安710069 [4]宁夏医科大学基础医学院生理学和神经生物学系,宁夏银川750001

出　　处：《生物工程学报》2023年第10期4085-4097,共13页Chinese Journal of Biotechnology

基　　金：陕西省重点研发计划(2022ZDLSF01-07);国家自然科学基金(81971560,82272343);国家“十三五”重大专项(2018ZX10302302002004)。

摘　　要：本研究旨在制备脂质体纳米颗粒(lipid nanoparticles,LNP)为载体的结核分枝杆菌(Mycobacterium tuberculosis,Mtb)抗原EsxV亚单位疫苗,明确该疫苗经黏膜免疫诱导的免疫应答水平。采用薄膜分散法制备包裹蛋白EsxV和c-di-AMP的LNP(EsxV:C:L),并对其包封率、LNP形态、粒径、表位电荷及多相分散指数进行了检测。EsxV:C:L滴鼻免疫BALB/c小鼠,检测免疫后血清和黏膜抗体、肺或脾细胞因子转录及分泌水平、肺T细胞亚群细胞比例。结果成功获得大小均一、呈球状、带负电的EsxV:C:L LNP亚单位疫苗。与EsxV:C相比,EsxV:C:L鼻黏膜接种可诱导小鼠呼吸道黏膜sIgA水平增加,脾细胞因子IL-2分泌水平升高,提高中央记忆T细和组织驻留T细胞的比例。综上,EsxV:C:L经黏膜免疫,可诱导更强的黏膜免疫和记忆性T细胞免疫应答,可能提供更好的抗Mtb感染的保护作用。To prepare a lipid nanoparticle(LNP)-based subunit vaccine of Mycobacterium tuberculosis(Mtb)antigen EsxV and study its immunological characteristics,the LNP containing EsxV and c-di-AMP(EsxV:C:L)was prepared by thin film dispersion method,and its encapsulation rate,LNP morphology,particle size,surface charge and polyphase dispersion index were measured.BALB/c mice were immunized with EsxV:C:L by nasal drops.The levels of serum and mucosal antibodies,transcription and secretion of cytokines in lung and spleen,and the proportion of T cell subsets were detected after immunization.EsxV:C:L LNPs were obtained with uniform size and they were spherical and negatively charged.Compared with EsxV:C immunization,EsxV:C:L mucosal inoculation induced increased sIgA level in respiratory tract mucosa.Levels of IL-2 secreted from spleen and ratios of memory T cells and tissue-resident T cells in mice were also elevated.In conclusion,EsxV:C:L could induce stronger mucosal immunity and memory T cell immune responses,which may provide better protection against Mtb infection.

关 键 词：结核分枝杆菌 EsxV 环二腺苷酸 脂质体纳米颗粒 亚单位疫苗 

分 类 号：R392[医药卫生—免疫学]