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首页> 外文期刊>World Journal of Gastroenterology >Viral replication modulated by synthetic peptide derived from hepatitis B virus X protein
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Viral replication modulated by synthetic peptide derived from hepatitis B virus X protein

机译：乙型肝炎病毒X蛋白衍生的合成肽调节病毒复制

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AIM: A strategy for viral vaccine design is the use of conserved peptides to overcome the problem of sequence diversity. At present it is still unclear whether conserved peptide is safe as a candidate vaccine. We reported it here for the first time not only to highlight the biohazard issue and safety importance for viral peptide vaccine, but also to explore the effect of a fully conserved peptide on HBV replication within the carboxyl terminus of HBx. METHODS: We synthesized the fully conserved peptide (CP) with nine residues, FVLGGCRHK. HBV-producing 2.2.15 cells were treated with or without 3.5 μM CP for 36 hours. Quantitative detection of viral DNA was performed by realtime PCR. HBV antigens were determined by enzyme-linked immunoadsorbent assay (ELISA). Quantitative analyses of p53 and Bax proteins were based on immunofluorescence. Flow cytometry was performed to detect cell cycle and apoptosis. RESULTS: Both extracellular and intracellular copies of HBV DNA per ml were significantly increased after incubation with 3.5 μM of CP. HBsAg and HBeAg in the cultured medium of CP-treatment cells were as abundant as untreated control cells. CP influenced negatively the extracellular viral gene products, and 3.5 μM CP could significantly inhibit intracellular HBsAg expression. In response to CP, intracellular HBeAg displayed an opposite pattern to that of HBsAg, and 3.5 μM CP could efficiently increase the level of intracellular HBeAg. Flow cytometric analyses exhibited no significant changes on cell cycle, apoptosis, p53 and Bax proteins in 2.2.15 cells with or without CP. CONCLUSION: Together with the results generated from the synthetic peptide, we address that the conserved region, a domain of HBx, may be responsible for modulating HBV replication. As conserved peptides from infectious microbes are used as immunogens to elicit immune responses, their latent biological hazard for human beings should be evaluated.
机译：目的：病毒疫苗设计的策略是使用保守的肽来克服序列多样性的问题。目前，尚不清楚保守肽作为候选疫苗是否安全。我们在此首次报道，不仅强调病毒肽疫苗的生物危害性问题和安全重要性，而且还探讨了完全保守的肽对HBx羧基末端内HBV复制的影响。方法：我们合成了具有9个残基的完全保守的肽（CP）FVLGGCRHK。在有或没有3.5μMCP的情况下，将产生HBV的2.2.15细胞处理36小时。通过实时PCR对病毒DNA进行定量检测。 HBV抗原通过酶联免疫吸附测定（ELISA）确定。 p53和Bax蛋白的定量分析基于免疫荧光。进行流式细胞仪检测细胞周期和凋亡。结果：与3.5μMCP孵育后，每毫升HBV DNA的细胞外和细胞内拷贝均显着增加。 CP处理细胞的培养基中的HBsAg和HBeAg与未处理的对照细胞一样丰富。 CP对细胞外病毒基因产物产生负面影响，而3.5μMCP可以显着抑制细胞内HBsAg表达。响应CP，细胞内HBeAg表现出与HBsAg相反的模式，而3.5μMCP可以有效增加细胞内HBeAg的水平。流式细胞仪分析显示，在有或没有CP的2.2.15细胞中，细胞周期，凋亡，p53和Bax蛋白没有显着变化。结论：与合成肽产生的结果一起，我们指出保守区域（HBx结构域）可能负责调节HBV复制。由于来自感染微生物的保守肽被用作引发免疫反应的免疫原，因此应评估其对人类的潜在生物危害。

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