Ubiquitin-modified hepatitis B virus core antigen effectively facilitates antigen presentation and enhances cytotoxic T lymphocyte activity via the cytoplasmic transduction peptide in vitro
Ubiquitin-modified hepatitis B virus core antigen effectively facilitates antigen presentation and enhances cytotoxic T lymphocyte activity via the cytoplasmic transduction peptide in vitro
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DOI：
10.3892/mmr.2015.3352
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发表时间：
2015-07-01
影响因子：
3.4
通讯作者：
Zang, Guoqing
中科院分区：
文献类型：
--
作者：
Song, Linlin;Zhuo, Meng;Zang, Guoqing
分化簇（CD）8（+）细胞毒性T淋巴细胞（CTL）在消除B型肝炎病毒（HBV）感染的细胞中具有关键作用。泛素（Ub）作为蛋白质降解的标志物起作用，其可促进适于主要组织相容性复合物I类呈递的肽的产生，而HBV核心抗原（HBcAg）具有显著的免疫原性特性。然而，UB修饰的HBcAg是否能够有效地诱导显著的CD 8（+）CTL活性仍有待阐明。为了解决这个问题，构建了原核表达载体，表达Ub-HBcAg-胞质转导肽（CTP）。融合蛋白成功表达，并随后脉冲到骨髓来源的树突状细胞（DC）。结果证实，在CTP的细胞穿透特性的帮助下，融合蛋白能够直接穿透到DC的细胞质中。结果表明，Ub-HBcAg-CTP融合蛋白不仅能增加DC表面分子的表达，促进增殖期T细胞分泌细胞因子，而且能诱导T细胞分化为特异性CTL，增强其抗病毒能力。结论：Ub-HBcAg-CTP融合蛋白能促进DC的成熟，增强靶向抗原的提呈，有效诱导HBcAg特异性CTL免疫应答。
Cluster of differentiation (CD)8(+) cytotoxic T lymphocytes (CTLs) have a key role in the elimination of hepatitis B virus (HBV)-infected cells. Ubiquitin (Ub) functions as a marker for protein degradation, which may promote the generation of peptides appropriate for major histocompatibility complex class I presentation, while the HBV core antigen (HBcAg) possesses marked immunogenic properties. However, it remains to be elucidated whether Ub-modified HBcAg is able to effectively elicit significant CD8(+) CTL activity. In order to address this issue, a prokaryotic vector was constructed to express the Ub-HBcAg-cytoplasmic transduction peptide (CTP). The fusion protein was successfully expressed and subsequently pulsed into bone-marrow-derived dendritic cells (DCs). It was confirmed that with assistance from the cell-penetrating properties of CTP, the fusion protein was able to directly penetrate into the cytoplasm of DCs. The results revealed that the Ub-HBcAg-CTP fusion protein not only increased the expression of surface molecules in DCs and cytokine secretion from proliferating T cells, but also induced T cells to differentiate into specific CTLs and enhanced their antiviral ability. In conclusion, the Ub-HBcAg-CTP fusion protein promoted DC maturation, enhanced the presentation of targeting antigens and efficiently induced HBcAg-specific CTL immune responses in vitro.