Improved Dendritic Cell-Based Immunization Against Hepatitis C Virus Using Peptide Inhibitors of Interleukin 10
Improved Dendritic Cell-Based Immunization Against Hepatitis C Virus Using Peptide Inhibitors of Interleukin 10
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DOI：
10.1002/hep.23980
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发表时间：
2011-01-01
期刊：
影响因子：
13.5
通讯作者：
Sarobe, Pablo
中科院分区：
文献类型：
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作者：
Diaz-Valdes, Nancy;Manterolal, Lorea;Sarobe, Pablo
高水平的白细胞介素10（IL-10）存在于慢性丙型肝炎病毒（HCV）感染已被认为是负责在这些患者中发现的抗病毒细胞免疫反应差。为了克服IL-10对抗原呈递细胞如树突状细胞（DC）的免疫抑制作用，我们开发了IL-10的肽抑制剂以恢复DC功能并伴随诱导有效的抗病毒免疫应答。使用噬菌体展示文库选择两种IL-10结合肽（p9和p13），并在生物测定和STAT-3（信号转导和转录激活因子3）磷酸化实验中评估它们抑制IL-10的能力。在HCV核心蛋白诱导IL-10产生的人白细胞培养物中，p13恢复了Toll样受体9（TLR9）刺激后浆细胞样DC产生干扰素α（IFN-α）的能力。类似地，当在HCV核心存在下用CD40L刺激髓样DC时，p9通过抑制HCV核心诱导的以及CD40L诱导的IL-10来增强IL-12的产生。此外，在体外，p13增强了成熟刺激对人和鼠DC的作用，增加了它们的IL-12产生和刺激活性，这导致响应T细胞的增殖和IFN-γ产生增强。最后，免疫与p13处理的小鼠DC诱导更强的抗HCV T细胞反应，不仅在野生型小鼠，但也在HCV转基因小鼠和在肝脏中瞬时表达HCV核心的小鼠。结论：这些结果表明，IL-10抑制肽可能具有重要的应用，以增强抗HCV免疫反应，通过恢复DC的免疫刺激能力。（肝脏学2011; 53：23 - 31）
The high levels of interleukin 10 (IL-10) present in chronic hepatitis C virus (HCV) infection have been suggested as responsible for the poor antiviral cellular immune responses found in these patients. To overcome the immunosuppressive effect of IL-10 on antigen-presenting cells such as dendritic cells (DCs), we developed peptide inhibitors of IL-10 to restore DC functions and concomitantly induce efficient antiviral immune responses. Two IL-10-binding peptides (p9 and p13) were selected using a phage-displayed library and their capacity to inhibit IL-10 was assessed in a bioassay and in STAT-3 (signal transducer and activator of transcription 3) phosphorylation experiments in vitro. In cultures of human leukocytes where HCV core protein induces the production of IL-10, p13 restored the ability of plasmacytoid DC to produce interferon alpha (IFN-alpha) after Toll-like receptor 9 (TLR9) stimulation. Similarly, when myeloid DCs were stimulated with CD40L in the presence of HCV core, p9 enhanced IL-12 production by inhibiting HCV core-induced as well as CD40L-induced IL-10. Moreover, in vitro, p13 potentiated the effect of maturation stimuli on human and murine DC, increasing their IL-12 production and stimulatory activity, which resulted in enhanced proliferation and IFN-gamma production by responding T-cells. Finally, immunization with p13-treated murine DC induced stronger anti-HCV T-cell responses not only in wildtype mice but also in HCV transgenic mice and in mice transiently expressing HCV core in the liver. Conclusion: These results suggest that IL-10 inhibiting peptides may have important applications to enhance anti-HCV immune responses by restoring the immunostimulatory capabilities of DC. (HEPATOLOGY 2011;53:23-31)