Generation of Immune Responses against Hepatitis C Virus by Dendritic Cells Containing NS5 Protein-Coated Microparticles
Generation of Immune Responses against Hepatitis C Virus by Dendritic Cells Containing NS5 Protein-Coated Microparticles
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DOI：
10.1128/cvi.00287-08
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发表时间：
2009-02-01
影响因子：
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通讯作者：
Wands, Jack R.
中科院分区：
文献类型：
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作者：
Gehring, Stephan;Gregory, Stephen H.;Wands, Jack R.
树突状细胞 (DC) 内化和处理抗原并激活细胞免疫反应。本研究的目的是确定含有抗原包被磁珠的 DC 诱导针对非结构性丙型肝炎病毒 (HCV) 抗原 5 (NS5) 的免疫力的能力。将源自 Fms 样酪氨酸激酶受体 3 (Flt3) 配体预处理的 BALB/c 小鼠的脾细胞与包被有 HCV NS5、脂多糖 (LPS) 和/或抗 CD40 的磁珠一起孵育；净化；并用于免疫接种。使用细胞毒性 T 淋巴细胞 (CTL) 和 T 细胞增殖测定、细胞内细胞因子染色以及体内表达 NS5 的 SP2/0 骨髓瘤细胞的同基因肿瘤攻击来测量细胞免疫。从接种含有 NS5、LPS 和抗 CD40 包被的珠子的 DC 的动物中分离的脾细胞在 NS5 存在的情况下分泌升高水平的白细胞介素 2 (IL-2) 和 γ 干扰素。在用含有涂有 NS5、LPS 和抗 CD40 的珠子的 DC 免疫的组中，产生 CD4(+)、IL-2 的细胞数量增加了 5 倍以上，同时脾细胞增殖反应也增强。与对照小鼠的水平相比，免疫将抗原特异性 CTL 活性提高三倍，并显着减少体内表达 NS5 的肿瘤细胞的生长。因此，采用 NS5 包被珠子的策略会在小鼠体内诱导细胞免疫反应，这与解决 HCV 的个体中发生的自然免疫反应密切相关。
Dendritic cells (DCs) internalize and process antigens as well as activate cellular immune responses. The aim of this study was to determine the capacity of DCs that contain antigen-coated magnetic beads to induce immunity against the nonstructural hepatitis C virus (HCV) antigen 5 (NS5). Splenocytes derived from Fms-like tyrosine kinase receptor 3 (Flt3) ligand-pretreated BALB/c mice were incubated with magnetic beads coated with HCV NS5, lipopolysaccharide (LPS), and/or anti-CD40; purified; and used for immunization. Cellular immunity was measured using cytotoxic T-lymphocyte (CTL) and T-cell proliferation assays, intracellular cytokine staining, and a syngeneic tumor challenge using NS5-expressing SP2/0 myeloma cells in vivo. Splenocytes isolated from animals vaccinated with DCs containing beads coated with NS5, LPS, and anti-CD40 secreted elevated levels of interleukin-2 (IL-2) and gamma interferon in the presence of NS5. The numbers of CD4(+), IL-2-producing cells were increased >5-fold in the group immunized with DCs containing beads coated with NS5, LPS, and anti-CD40, paralleled by an enhanced splenocyte proliferative response. Immunization promoted antigen-specific CTL activity threefold compared to the level for control mice and significantly reduced the growth of NS5-expressing tumor cells in vivo. Thus, strategies that employ NS5-coated beads induce cellular immune responses in mice, which correlate well with the natural immune responses that occur in individuals who resolve HCV.