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首页> 外文期刊>World Journal of Gastroenterology >Development of hepatitis C virus vaccine using hepatitis B core antigen as immuno-carrier.
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Development of hepatitis C virus vaccine using hepatitis B core antigen as immuno-carrier.

机译：以乙型肝炎核心抗原为免疫载体的丙型肝炎病毒疫苗的研制。

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AIM: To develop hepatitis C virus (HCV) vaccine using HBcAg as the immuno-carrier to express HCV T epitope and to investigate its immunogenicity in mice. METHODS: We constructed the plasmid pTrc-core(NheI) using gene engineering technique, constructed the pcDNA3.1-core(NheI)-GFP plasmid with GFP as the reporter gene, and transfected them into Hela cells. The expression of GFP was observed under confocal microscopy and the feasibility of using HBcAg as an immuno-carrier vaccine was studied. pTrc-core gene with a synthetic T epitope antigen gene of HCV (35-44aa) was fused and expressed in the plasmid pTrc-core-HCV (T). For the fusion of the HBcAg-T protein, sucrose, density gradient centrifugation was used, and its molecular weight and purity were analyzed by SDS-PAGE. Then balb/c mice were immunized by the plasmid with the HBcAg (expressed by pTrc-core) protein as control. The tumor regression potential was investigated in mice and evaluated at appropriate time. After three times of immunization, the peripheral blood and spleen of vaccinated mice were collected. HBcAb was detected by ELISA, and nonspecific T lymphocyte proliferation and response of splenocytes were respectively examined by MTT assay. T cell subset of blood and spleen were detected by FACS. RESULTS: GFP was successfully expressed. Tumor regression trial showed that no tumor formation was found in the group receiving immunization, while tumor xenograft progression was not changed in the control group. Strong nonspecific lymphocyte proliferation response was induced. FACS also showed that the ratio of CD8(+) T cells in the experimental group was higher than the controls, but the serum HBcAb in experimental group was similar to the control. CONCLUSION: HBcAg can be used as an immuno-carrier of vaccine, the fusion of HBcAg-T protein could induce stronger cellular immune responses and it might be a candidate for therapeutic vaccines specific for HCV.
机译：目的：开发以HBcAg为免疫载体表达HCV T表位的丙型肝炎病毒疫苗，并研究其在小鼠中的免疫原性。方法：利用基因工程技术构建了质粒pTrc-core（NheI），构建了以GFP为报告基因的pcDNA3.1-core（NheI）-GFP质粒，并将其转染到Hela细胞中。在共聚焦显微镜下观察GFP的表达，并研究了使用HBcAg作为免疫载体疫苗的可行性。将具有合成的HCV T表位抗原基因的pTrc-core基因（35-44aa）融合并在质粒pTrc-core-HCV（T）中表达。对于蔗糖HBcAg-T蛋白的融合，使用密度梯度离心，并通过SDS-PAGE分析其分子量和纯度。然后，以HBcAg（由pTrc-核心表达）蛋白作为对照，通过质粒免疫balb / c小鼠。在小鼠中研究了肿瘤消退潜力，并在适当的时间进行了评估。免疫三遍后，收集接种小鼠的外周血和脾脏。 ELISA法检测HBcAb，MTT法分别检测非特异性T淋巴细胞增殖和脾细胞反应。通过FACS检测血液和脾的T细胞亚群。结果：GFP成功表达。肿瘤消退试验显示，在接受免疫的组中未发现肿瘤形成，而对照组中异种移植的进展没有变化。诱导强烈的非特异性淋巴细胞增殖反应。 FACS还显示实验组中CD8（+）T细胞的比例高于对照组，但实验组中的血清HBcAb与对照组相似。结论：HBcAg可作为疫苗的免疫载体，HBcAg-T蛋白的融合可诱导更强的细胞免疫反应，可能是HCV特异治疗性疫苗的候选者。

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