A novel TLR7 agonist as adjuvant to stimulate high quality HBsAg-specific immune responses in an HBV mouse model
A novel TLR7 agonist as adjuvant to stimulate high quality HBsAg-specific immune responses in an HBV mouse model
复制标题
一种新型 TLR7 激动剂作为佐剂在 HBV 小鼠模型中刺激高质量 HBsAg 特异性免疫反应
DOI：
10.1186/s12967-020-02275-2
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发表时间：
2020-03-04
影响因子：
7.4
通讯作者：
Jin, Guangyi
中科院分区：
文献类型：
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作者：
Hu, Yunlong;Tang, Li;Jin, Guangyi
背景B型肝炎病毒（HBV）感染的发病率和死亡率在全球范围内是巨大的。新的治疗方法，可以诱导保护性免疫反应，迫切需要有效地控制HBV流行，并最终根除慢性HBV infection.MethodsWe设计和评估的HBV治疗性疫苗组成的一种新的Toll样受体7（TLR 7）激动剂T7-EA，明矾佐剂和重组HBsAg蛋白。我们使用RNA-seq、ELISA和hTLR 7/8报告分析来体外表征T7-EA，并使用实时PCR来评估体内组织保留特性。结果T7-EA对Raw 264.7细胞具有特异性的hTLR 7受体激动剂作用，并诱导了与未修饰的TLR 7配体相似的基因表达模式;然而，T7-EA比未修饰的TLR 7配体更有效。体内研究表明，T7-EA具有组织保留活性，刺激局部细胞因子和趋化因子表达长达7天。T7-EA可诱导Th 1型免疫应答，表现为与接受传统明矾佐剂HBV疫苗的小鼠相比，在正常小鼠中增加了HBsAg特异性IgG 2a滴度和T细胞应答。结论T7-EA可作为HBV疫苗的候选佐剂。
BackgroundThe global burden of hepatitis B virus (HBV) infection in terms of morbidity and mortality is immense. Novel treatments that can induce a protective immune response are urgently needed to effectively control the HBV epidemic and eventually eradicate chronic HBV infection.MethodsWe designed and evaluated an HBV therapeutic vaccine consisting of a novel Toll-like receptor 7 (TLR7) agonist T7-EA, an Alum adjuvant and a recombinant HBsAg protein. We used RNA-seq, ELISA and hTLR7/8 reporting assays to characterize T7-EA in vitro and real-time PCR to evaluate the tissue-retention characteristics in vivo. To evaluate the adjuvant potential, we administrated T7-EA intraperitoneally in a formulation with an Alum adjuvant and HBsAg in normal and HBV mice, then, we evaluated the HBsAg-specific immune responses by ELISA and Elispot assays.ResultsT7-EA acted as an hTLR7-specific agonist and induced a similar gene expression pattern as an unmodified TLR7 ligand when Raw 264.7 cells were exposed to T7-EA; however, T7-EA was more potent than the unmodified TLR7 ligand. In vivo studies showed that T7-EA had tissue-retaining activity with stimulating local cytokine and chemokine expression for up to 7 days. T7-EA could induce Th1-type immune responses, as evidenced by an increased HBsAg-specific IgG2a titer and a T-cell response in normal mice compared to mice received traditional Alum-adjuvant HBV vaccine. Importantly, T7-EA could break immune tolerance and induce persistent HBsAg-specific antibody and T-cell responses in an HBV mouse model.ConclusionsT7-EA might be a candidate adjuvant in a prophylactic and therapeutic HBV vaccine.