酵母微囊封装的纳米乳佐剂增强亚单位疫苗的体液和细胞免疫应答  

MF59 and subunit vaccine encapsulated in yeast microcapsules with enhanced humoral and cellular immune responses

作　　者：章婉娴 黄婷婷 吴昌静 张远冬 ZHANG Wan-xian;HUANG Ting-ting;WU Chang-jing;ZHANG Yuan-dong(School of Pharmacy,Zunyi Medical University,Zunyi 563000,China)

机构地区：[1]遵义医科大学药学院,贵州遵义563000

出　　处：《药学学报》2025年第4期1019-1028,共10页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(82160678,82003686);中国博士后科学基金(2019M650249,2020T130088ZX)。

摘　　要：利用酵母衍生的微囊多级装载纳米乳佐剂MF59和抗原,以解决MF59佐剂不能直接包载抗原以及诱导细胞免疫能力不足的问题。利用强酸碱法制备酵母微囊(yeast microcapsules,YC),其具有多孔、中空的结构和较好的佐剂效应。将聚合物聚己内酯-聚乙烯亚胺(polycaprolactone-polyethyleneimine,PCL-PEI)修饰MF59纳米乳获得正电性的乳粒,通过静电力驱动使乳粒自发沉积到YC中,并进一步吸附抗原鸡卵清白蛋白(ovalbumin,OVA),得到共载抗原和佐剂的疫苗YC-MF59-OVA。YC-MF59-OVA基于囊壳上β-葡聚糖可被抗原提呈细胞(antigen presenting cells,APCs)识别而内吞,同时在接种部位募集抗原提呈细胞,进而有效激活机体免疫响应,使血清中IgG、IgG1、IgG2a抗体水平相比游离OVA升高了2~3倍;诱导了显著高于OVA的IFN-γ^(+)CD8^(+)、IL-4^(+)CD4^(+)T细胞免疫应答,刺激脾组织中记忆性CD44^(+)CD62L^(+)T细胞比例显著升高。本动物实验方案经遵义医科大学实验动物伦理委员会审核批准(批准号:ZMU21-2407-169)。研究结果表明,YC-MF59-OVA能够激活强效的体液免疫应答和细胞免疫应答,证实YC能够显著弥补纳米乳佐剂的不足,为后续疫苗递送系统的开发提供新的参考。Yeast-derived microcapsules were employed to co-encapsulate a nano-emulsion adjuvant (MF59) and antigens,effectively addressing the limitations of MF59 adjuvant in direct antigen encapsulation and its capacity to induce cellular immunity.Yeast microcapsules (YCs) were prepared using strong acid and alkali treatments,resulting in a porous and hollow structure with enhanced adjuvant properties.Positively charged polycaprolactone-polyethyleneimine (PCL-PEI) modified MF59 nanoemulsions were produced,which allowed for electrostatic interaction-driven spontaneous deposition into YCs.This modification facilitated the adsorption of the antigen,chicken ovalbumin (OVA),forming the complex YC-MF59-OVA.YC-MF59-OVA was efficiently recognized and endocytosed by antigen-presenting cells (APCs),a process facilitated by the β-glucan present on the capsular shell.Simultaneously,YC-MF59-OVA enabled a sustained release of the antigen and promoted the recruitment of APCs at the site of inoculation,leading to enhanced activation of immune responses in mice.Specifically,YC-MF59-OVA significantly elevated serum levels of IgG,IgG1,and IgG2a antibodies,achieving concentrations that were two to three times higher than those observed in the group treated with free OVA.In addition,the cellular immune response was notably improved,as evidenced by increased frequencies of IFN-γ~+CD8~+ and IL-4~+CD4~+T cells compared to the OVA-immunized group.Furthermore,there was a marked increase in the proportion of memory T cells (CD44~+CD62L~+) in the splenic tissues of treated animals.The animal experiment protocol was reviewed and approved by Institutional Animal Care and Use Committee of Zunyi Medical University (approval No.ZMU21-2407-169).These findings demonstrate that YC-MF59-OVA can elicit robust humoral and cellular immune responses,confirming that YC can significantly overcome the limitations of traditional nanoemulsion adjuvants.This study provides a promising reference for the development of advanced vaccine delivery systems.

关 键 词：酵母微囊 纳米乳 免疫佐剂 亚单位疫苗 细胞免疫应答 

分 类 号：R966[医药卫生—药理学]