PDF(Pubmed)
Abstract：
Hepatitis B virus (HBV) infection is a major public health concern. The clearance of HBV may involve cytotoxic T-lymphocyte (CTL) activity and T helper type 1 reactions. Exosomes generated from dendritic cells (DCs) can induce immunological responses capable of eradicating viruses. However, exosomes loaded with antigens have not yet demonstrated therapeutic potential in HBV infection. Therefore, the present study aimed to investigate the antiviral effects of DC-derived exosomes (Dexs) loaded with ubiquitinated HBV core antigen (Dexs-Ub-HBcAg). Murine bone marrow-derived DCs were loaded with a recombinant lentivector encoding the ubiquitinated form of HBcAg. High-purity Dexs were generated using differential velocity centrifugation. Splenic T-lymphocytes were stimulated with Dexs-Ub-HBcAg and the specific T-cell-mediated immune responses were examined. Cytokine expression was analyzed using enzyme-linked immunosorbent assays. T-lymphocyte proliferation was detected using a Cell Counting Kit-8 assay and HBcAg-specific CTL activity was determined using a lactate dehydrogenase release assay. The results revealed that Dexs-Ub-HBcAg effectively stimulated T-cell proliferation and induced the activation of antigen-specific CTLs to exhibit HBcAg-specific CTL immune responses in vitro. These results suggest the potential of Dexs-Ub-HBcAg for development as a future therapeutic option for the elimination of HBV.
摘要：
乙型肝炎病毒（HBV）感染是主要的公共卫生问题。HBV的清除可能涉及细胞毒性T淋巴细胞（CTL）活性和T辅助1型反应。由树突状细胞(DC)产生的外来体可以诱导能够根除病毒的免疫应答。然而,载有抗原的外泌体尚未证明在HBV感染中的治疗潜力。因此,本研究旨在研究负载泛素化HBV核心抗原（Dexs-Ub-HBcAg）的DC衍生外泌体（Dexs）的抗病毒作用。用编码HBcAg的泛素化形式的重组lentivector装载鼠骨髓来源的DC。使用差速离心产生高纯度Dexs。用Dexs-Ub-HBcAg刺激脾T淋巴细胞，并检查特异性T细胞介导的免疫反应。使用酶联免疫吸附测定分析细胞因子表达。使用细胞计数试剂盒-8测定法检测T淋巴细胞增殖,并使用乳酸脱氢酶释放测定法测定HBcAg特异性CTL活性。结果表明，Dexs-Ub-HBcAg可有效刺激T细胞增殖，并诱导抗原特异性CTL的激活，从而在体外表现出HBcAg特异性CTL免疫反应。这些结果表明Dexs-Ub-HBcAg作为消除HBV的未来治疗选择的发展潜力。
