An Engineered DC-Targeting Lentivector Induces Robust T Cell Responses and Inhibits HBV Replication in HBV Transgenic Mice via Upregulating T Cell Autophagy
An Engineered DC-Targeting Lentivector Induces Robust T Cell Responses and Inhibits HBV Replication in HBV Transgenic Mice via Upregulating T Cell Autophagy
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工程化的 DC 靶向慢载体通过上调 T 细胞自噬在 HBV 转基因小鼠中诱导强大的 T 细胞反应并抑制 HBV 复制
DOI：
10.1159/000491972
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发表时间：
2018-07
影响因子：
--
通讯作者：
Zhenghao Tang
中科院分区：
文献类型：
--
作者：
Siyuan Ma;Xiaohua Chen;Quanhui Tan;Shenglan Dai;Dan Li;Shanshan Wu;Yongsheng Yu;Guoqing Zang;Zhenghao Tang
背景/目标：开发工程化的树突状细胞（DC）靶向慢病毒载体（LV）已经成为深入研究的目标，因为它们有可能产生可以安全地施用于患者的抗原导向的免疫治疗剂。在这项研究中，我们构建了一个DC导向的LV（LVDC-UbHBcAg-LIGHT）作为一个潜在的疫苗，以诱导抗HBV免疫应答。方法：通过活体动物成像研究证实LVDC-UbHBcAg-LIGHT对体内DC的特异性。通过流式细胞术评估T细胞中细胞因子产生的水平。LDH释放法和ELISA法分别检测LVs诱导的HBcAg特异性细胞毒性T淋巴细胞（CTL）应答和抗体应答。分别用Abbott试剂盒和定量PCR检测血清HBsAg和HBV DNA水平。免疫组化法检测HBV转基因小鼠肝组织中HBsAg和HBcAg的表达水平。此外，还探讨了CD 8 + T细胞活化的分子机制。结果如下：活体动物成像研究显示，LVDC-UbHBcAg-LIGHT皮下给药后，在注射部位未检测到明显的发光信号。用LVDC-UbHBcAg-LIGHT免疫在HBV转基因小鼠中引起有效的T细胞应答，表现为产生IFN-γ、TNF-α和GzmB的CD 8 + T细胞以及产生IFN-γ的CD 4 + T细胞的百分比增加，HBcAg特异性CTL活性和抗体应答改善。此外，用LVDC-UbHBcAg-LIGHT接种有效地降低了HBV转基因小鼠的血清HBsAg、HBV DNA水平以及肝组织中HBsAg和HBcAg的表达。更重要的是，在活化的CD 8 + T细胞中诱导自噬，并且诱导的自噬通过选择性降解泛素化的凋亡和细胞周期相关蛋白聚集体而显著促进T细胞的增殖并降低凋亡的CD 8 + T细胞的频率。此外，我们通过共聚焦显微镜和免疫沉淀分析证实了自噬体和泛素化聚集体之间的相互作用。结论：这些结果表明，LVDC-UbHBcAg-LIGHT提供了一种简单的方法，在HBV转基因小鼠中引发有效的抗病毒免疫应答，并可能作为一种潜在的治疗策略，以更安全和更有效地根除HBV。此外，我们的研究结果揭示了自噬在促进活化的CD 8 + T细胞的存活和增殖中的直接作用。
Background/Aims: Developing engineered dendritic cell (DC)-targeting lentivectors (LVs) have been the target of intense research for their potential to create antigen-directed immunotherapeutics which can be safely administered to patients. In this study, we constructed a DC-directed LV (LVDC-UbHBcAg-LIGHT) as a potential vaccine to induce anti-HBV immune responses. Methods: Specificity of LVDC-UbHBcAg-LIGHT for DCs in vivo was confirmed through live animal imaging studies. The levels of cytokine production in T cells were assessed by flow cytometry. The HBcAg-specific cytotoxic T lymphocyte (CTL) responses and antibody responses induced by direct administration of the LVs were detected by LDH release assay and ELISA respectively. The levels of serum HBsAg and HBV DNA were evaluated by Abbott kits and quantitative PCR respectively. The expression levels of HBsAg and HBcAg in liver tissues of HBV transgenic mice were examined by immunohistochemistry. In addition, molecular mechanism underlying the activation of CD8+ T cells was explored. Results: Live animal imaging studies showed that following subcutaneous administration of LVDC-UbHBcAg-LIGHT, no obvious luminescence signal was detected at the injection site. Immunization with LVDC-UbHBcAg-LIGHT elicited potent T cell responses in HBV transgenic mice evidenced by increased percentages of IFN-γ, TNF-α and GzmB producing CD8+ T cells as well as IFN-γ producing CD4+ T cells, improved HBcAg-specific CTL activities and antibody responses. Additionally, vaccination with LVDC-UbHBcAg-LIGHT efficiently reduced serum HBsAg, HBV DNA levels and the expression of HBsAg and HBcAg in liver tissues of HBV transgenic mice. More importantly, autophagy was induced in the activated CD8+ T cells, and the induced autophagy noticeably promoted the proliferation of T cells and decreased the frequencies of apoptotic CD8+ T cells by selectively degrading ubiquitinated apoptosis and cell cycle-associated protein aggregates. Futhermore, we confirmed the interaction between autophagosomes and ubiquitinated aggregates by confocal microscopy and immunoprecipitation analysis. Conclusions: These results demonstrated that LVDC-UbHBcAg-LIGHT provided a simple method of eliciting effective antiviral immune responses in HBV transgenic mice and might potentially be used as a therapeutic strategy to eradicate HBV with more safety and efficiency. Moreover, our results revealed a direct role of autophagy in promoting the survival and proliferation of activated CD8+ T cells.
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DOI：
10.1084/jem.20061303
发表时间：
2007-01-22
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Pua HH;Dzhagalov I;Chuck M;Mizushima N;He YW
通讯作者：
He YW
影响因子：
64.5
作者：
Geijtenbeek, TBH;Torensma, R;Figdor, CG
通讯作者：
Figdor, CG
DOI：
10.1172/jci17098
发表时间：
2003-06
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
C. Esslinger;Laurence Chapatte;D. Finke;I. Miconnet;P. Guillaume;F. Lévy;H. Macdonald
通讯作者：
C. Esslinger;Laurence Chapatte;D. Finke;I. Miconnet;P. Guillaume;F. Lévy;H. Macdonald
DOI：
10.4049/jimmunol.1502061
发表时间：
2016-04-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Zhu D;Liu L;Yang D;Fu S;Bian Y;Sun Z;He J;Su L;Zhang L;Peng H;Fu YX
通讯作者：
Fu YX
影响因子：
5.4
作者：
GUIDOTTI, LG;MATZKE, B;CHISARI, FV
通讯作者：
CHISARI, FV