鉴定影响乙型肝炎病毒小包膜蛋白降解的病毒学因素

张宝予; 王勇翔

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微生物与感染 ›› 2025, Vol. 20 ›› Issue (1) : 1-8.

论著

鉴定影响乙型肝炎病毒小包膜蛋白降解的病毒学因素

张宝予¹,王勇翔²

作者信息 +

Identification of virological factors affecting the degradation of hepatitis B virus small envelope protein

Bao-Yu ZHANG¹,Yong-Xiang Wang

Author information +

文章历史 +

摘要

乙型肝炎病毒（hepatitis B virus，HBV）的包膜包含大、中、小包膜蛋白（large, middle, small envelope protein; L, M, S）。当前慢性乙型肝炎的治疗目标是功能性治愈，其关键指标是外周血中的乙肝表面抗原（hepatitis B surface antigen, HBsAg）持续消失。但目前关于HBsAg降解的机制仍不明确。我们在前期工作中发现，靶向HBV包膜蛋白的嵌合抗原受体（chimeric antigen receptor, CAR）G12-CAR抑制HBsAg分泌，并促进其在胞内降解。为研究相关机制，本研究采用多种表达载体转染Huh-7人肝癌细胞表达S蛋白，再用巴佛洛霉素（bafilomycin）和硼替佐米（bortezomib）分别抑制细胞内的溶酶体自噬和蛋白酶体降解途径，并通过Western blot和酶联免疫吸附试验检测细胞内和外的S蛋白。结果显示，S蛋白可通过溶酶体自噬和蛋白酶体途径降解；L蛋白可干扰S蛋白在细胞内降解；编码S蛋白的mRNA的 3’非翻译区（untranslated region, UTR）序列也影响了S蛋白在细胞内降解。

Abstract

The envelope proteins of hepatitis B virus can be classified into large envelope protein (L), middle envelope protein (M) and small envelope protein (S), and S protein is the most abundant transmembrane protein in the viral envelope. The current treatment goal for chronic hepatitis B is functional cure. One of the important indicators of functional cure is the sustained disappearance of hepatitis B surface antigen (HBsAg) in the blood, but the mechanism of intracellular degradation of S protein is still unclear. In our previous work, we found that G12-CAR, a chimeric antigen receptor (CAR) targeting hepatitis B virus envelope proteins, inhibited S protein secretion and promoted its intracellular degradation. In this study, Huh-7 human hepatocellular carcinoma cells were transfected with different vectors to express S protein, and bafilomycin and bortezomib were used to inhibit lysosomal autophagy and proteasomal degradation. The intracellular expression products were identified using SDS-PAGE and Western blot, and the extracellular expression products were detected by ELISA. The results showed that S proteins could be degraded by lysosomal autophagy and proteasome pathways; some hepatitis B virus factors affected S protein degradation; L proteins affected intracellular degradation of S proteins; and the envelope protein mRNA 3'-UTR sequences in the HBV genome affected intracellular S protein degradation.

导出引用

张宝予王勇翔. 鉴定影响乙型肝炎病毒小包膜蛋白降解的病毒学因素[J]. 微生物与感染. 2025, 20(1): 1-8

Bao-Yu ZHANG Yong-Xiang Wang. Identification of virological factors affecting the degradation of hepatitis B virus small envelope protein[J]. Journal of Microbes and Infections. 2025, 20(1): 1-8

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