Liu Shang, Wang Jie, Li Yunxuan, Wang Muhan, Du Pei, Zhang Zhijie, Li Wenguo, Sun Rongchen, Fan Mingtao, Yang Meijia, Yin Hongping
School of Life Science and Technology, China Pharmaceutical University, Nanjing 211198, China.
Jiangsu Cell Tech Medical Research Institute Co., Ltd., Nanjing 211100, China.
Pharmaceutics. 2025 Feb 7;17(2):211. doi: 10.3390/pharmaceutics17020211.
: In chronic hepatitis B infection (CHB), the hepatitis B surface antigen (HBsAg) continuously exhausts the hepatitis B surface antibody (HBsAb), which leads to the formation of immune tolerance. Accordingly, the hepatitis B virus (HBV) infection can be blocked by inhibiting the binding of the hepatitis B surface pre-S1/pre-S2 antigen to the hepatocyte receptor NTCP, but the clinical cure rate of pre-S-based vaccines for CHB is limited. : In this study, we designed and prepared multivalent hepatitis B therapeutic mRNA vaccines encoding three hepatitis B surface antigen proteins (L, M, and S) at the cell membrane, verified via in vitro transfection and expression experiments. An in vivo immunization experiment in HBV transgenic (Tg) mice was first completed. Subsequently, an adeno-associated virus plasmid vector carrying the HBV1.2-fold genome (pAAV HBV1.2) model and the adeno-associated virus vector carrying HBV1.3-fold genome (rAAV HBV1.3) model were constructed and immunized with mRNA vaccines. The HBV antigen, antibodies, and HBV DNA in serum were detected. Indirect (enzyme-linked immunosorbent assay) ELISA were made to analyze the activated antigen-specific IgG in HBV Tg mice. Antigen-dependent T-cell activation experiments were carried out, as well as the acute toxicity tests in mice. : The L protein/pre-S antigens could be stably presented at the cell membrane with the support of the S protein (and M protein). After vaccinations, the vaccines effectively reactivated the production of high levels of HBsAb, disrupted immune tolerance, and activated the production of high-affinity antibodies against structural pre-S antigen in HBV Tg mice. The HBsAg seroconversion and serum HBV DNA clearance were achieved in two HBV mice models. Furthermore, pre-S antigen-dependent T-cell response against HBV infection was confirmed. The therapeutic vaccine also showed safety in mice. : A novel therapeutic mRNA vaccine was developed to break through HBsAg-mediated immune tolerance and treat CHB by stably presenting the pre-S antigen at the membrane, and the vaccine has great potential for the functional cure of CHB.
在慢性乙型肝炎感染（CHB）中，乙型肝炎表面抗原（HBsAg）持续消耗乙型肝炎表面抗体（HBsAb），从而导致免疫耐受的形成。因此，通过抑制乙型肝炎表面前S1/前S2抗原与肝细胞受体NTCP的结合，可以阻断乙型肝炎病毒（HBV）感染，但基于前S的CHB疫苗的临床治愈率有限。在本研究中，我们设计并制备了在细胞膜上编码三种乙型肝炎表面抗原蛋白（L、M和S）的多价乙型肝炎治疗性mRNA疫苗，并通过体外转染和表达实验进行了验证。首先在HBV转基因（Tg）小鼠中完成了体内免疫实验。随后，构建了携带HBV 1.2倍基因组的腺相关病毒质粒载体（pAAV HBV1.2）模型和携带HBV 1.3倍基因组的腺相关病毒载体（rAAV HBV1.3）模型，并用mRNA疫苗进行免疫。检测血清中的HBV抗原、抗体和HBV DNA。采用间接（酶联免疫吸附测定）ELISA分析HBV Tg小鼠中活化的抗原特异性IgG。进行了抗原依赖性T细胞活化实验以及小鼠急性毒性试验。L蛋白/前S抗原在S蛋白（和M蛋白）的支持下可以稳定地呈现在细胞膜上。接种疫苗后，疫苗有效地重新激活了高水平HBsAb的产生，打破了免疫耐受，并激活了HBV Tg小鼠中针对结构性前S抗原的高亲和力抗体的产生。在两种HBV小鼠模型中实现了HBsAg血清学转换和血清HBV DNA清除。此外，证实了针对HBV感染的前S抗原依赖性T细胞反应。该治疗性疫苗在小鼠中也显示出安全性。开发了一种新型治疗性mRNA疫苗，通过在膜上稳定呈递前S抗原突破HBsAg介导的免疫耐受并治疗CHB，该疫苗在CHB的功能性治愈方面具有巨大潜力。
Zotero 插件
