Oral Combination Vaccine, Comprising Bifidobacterium Displaying Hepatitis C Virus Nonstructural Protein 3 and Interferon-α, Induces Strong Cellular Immunity Specific to Nonstructural Protein 3 in Mice
Oral Combination Vaccine, Comprising Bifidobacterium Displaying Hepatitis C Virus Nonstructural Protein 3 and Interferon-α, Induces Strong Cellular Immunity Specific to Nonstructural Protein 3 in Mice
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口服组合疫苗包含展示丙型肝炎病毒非结构蛋白 3 的双歧杆菌和干扰素-α，可在小鼠中诱导针对非结构蛋白 3 的强细胞免疫
DOI：
10.1089/vim.2016.0111
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发表时间：
2017
期刊：
影响因子：
2.2
通讯作者：
白川利朗
中科院分区：
文献类型：
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作者：
北川孝一;大本知佳;小田麗未;荒木綾芽;斉藤大樹;重村克巳;片山高嶺;堀田博;白川利朗
我们先前利用展示丙型肝炎病毒非结构蛋白3（NS 3）多肽的双歧杆菌制备了一种口服丙型肝炎病毒（HCV）疫苗。NS 3特异性细胞免疫对于病毒清除和从HCV感染中恢复是重要的。在本研究中，我们增强了口服HCV疫苗长双歧杆菌2165（B）诱导的细胞免疫应答。longum 2165），通过在小鼠实验模型中将作为佐剂的干扰素-α（IFN-α）与疫苗组合。IFN-α是一种广泛使用的细胞因子，符合HCV感染的标准护理（SOC），并发挥各种免疫调节作用。用B. longum 2165隔日给药和/或IFN-α每周两次给药，持续一个月，然后使用脾细胞分析免疫应答。我们通过细胞因子定量、细胞内细胞因子染色和靶向表达NS 3/4A蛋白的EL 4肿瘤细胞（EL 4-NS 3/4A）的细胞毒性T淋巴细胞（CTL）测定来确定NS 3特异性细胞免疫的诱导。我们还对荷EL 4-NS 3/4A肿瘤的小鼠进行了体内联合治疗。结果证实，B.与磷酸盐缓冲液对照组（B）相比，longum 2165和IFN-α诱导的IFN-γ分泌量、分泌IFN-γ的CD 4 +T细胞和CD 8 +T细胞数量以及对EL 4-NS 3/4 A细胞的CTL活性均显著增加。longum 2165单药组与IFN-α单药组比较，差异有显著性（p< 0.05）。我们还证实了联合治疗强烈增强了体内肿瘤生长抑制作用，而没有严重的不良反应（p< 0.05）。这些结果表明，B. longum 2165和IFN-α联合治疗可诱导针对NS 3蛋白的强细胞免疫，增强目前针对慢性HCV感染的SOC免疫治疗。
We previously generated an oral hepatitis C virus (HCV) vaccine usingBifidobacteriumdisplaying the HCV nonstructural protein 3 (NS3) polypeptide. NS3-specific cellular immunity is important for viral clearance and recovery from HCV infection. In this study, we enhanced the cellular immune responses induced by our oral HCV vaccine,Bifidobacterium longum2165 (B. longum2165), by combining interferon-α (IFN-α) as an adjuvant with the vaccine in a mouse experimental model. IFN-α is a widely used cytokine meeting the standard of care (SOC) for HCV infection and plays various immunoregulatory roles. We treated C57BL/6N mice withB. longum2165 every other day and/or IFN-α twice a week for a month and then analyzed the immune responses using spleen cells. We determined the induction of NS3-specific cellular immunity by cytokine quantification, intracellular cytokine staining, and a cytotoxic T lymphocyte (CTL) assay targeting EL4 tumor cells expressing NS3/4A protein (EL4-NS3/4A). We also treated mice bearing EL4-NS3/4A tumor with the combination therapyin vivo. The results confirmed that the combination therapy ofB. longum2165 and IFN-α induced significantly higher IFN-γ secretion, higher population of CD4+T and CD8+T cells secreting IFN-γ, and higher CTL activity against EL4-NS3/4A cells compared with the control groups of phosphate-buffered saline,B. longum2165 alone, and IFN-α alone (p< 0.05). We also confirmed that the combination therapy strongly enhanced tumor growth inhibitory effectsin vivowith no serious adverse effects (p< 0.05). These results suggest that the combination ofB. longum2165 and IFN-α could induce a strong cellular immunity specific to NS3 protein as a combination therapy augmenting the current SOC immunotherapy against chronic HCV infection.