Medical Heredity Research Center, Northwest Women's and Children's Hospital, Shaanxi, PR China.
Medical Heredity Research Center, Northwest Women's and Children's Hospital, Shaanxi, PR China.
Mol Immunol. 2021 Feb;130:7-13. doi: 10.1016/j.molimm.2020.12.004. Epub 2020 Dec 16.
Targeted therapy for patients with hepatitis B virus (HBV) infection can lead to objective responses, although response times may be short. At the same time, the response rate to programmed cell death-1 (PD-1) treatment was more durable. It is speculated that HBV targeted therapy can synergistically enhance the antitumor activity with PD-1 blockade. To test this hypothesis, we evaluated the effect of crispr-cas9 on HBV and PD-1 in vitro and in vivo. We found that HBV targeting gRNA/cas9 induced a decrease in the expression of HBsAg, while the PD-1 gene could be knocked out by electroporation targeting gRNA / cas9 by polymerase chain reaction. In HBV transgenic mice, the immunophenotype and cytokine expression of human dendritic cells (DCS) were detected by crispr-cas9 system stimulation, flow cytometry and polymerase chain reaction. These results indicate that gRNA/cas9 treatment upregulates the expression of CD80, CD83 and CD86, and significantly increases the mRNA levels of IL-6, IL-12, IL-23 and tumor necrosis factor alpha. The combination of anti HBV and anti PD-1 therapy can inhibit HBV expression and significantly improve the survival of HBV transgenic mice. In addition, the combination therapy increased the production of interferon by T cells, and then enhanced the expression of Th1 related immunostimulatory genes, thereby reducing the transcription of regulatory / inhibitory immune genes. In general, this response can reshape the tumor microenvironment from immunosuppression to immune stimulation. Finally, anti HBV therapy can induce the expression of interferon dependent programmed cell death ligand-1 in HBV transgenic mice in vivo. To sum up, these results demonstrate that the combination of HBV targeted therapy and PD-1 immune checkpoint block has a strong synergistic effect, thus supporting the transformation potential of this combined therapy strategy in clinical treatment of HBV infection.
针对乙型肝炎病毒 (HBV) 感染患者的靶向治疗可导致客观应答，尽管应答时间可能较短。同时，程序性细胞死亡-1 (PD-1) 治疗的应答率更为持久。人们推测 HBV 靶向治疗可以与 PD-1 阻断协同增强抗肿瘤活性。为了验证这一假设，我们在体外和体内评估了 crispr-cas9 对 HBV 和 PD-1 的作用。我们发现，HBV 靶向 gRNA/cas9 诱导 HBsAg 表达降低，而通过靶向 gRNA / cas9 的电穿孔可以通过聚合酶链反应敲除 PD-1 基因。在 HBV 转基因小鼠中，通过 crispr-cas9 系统刺激、流式细胞术和聚合酶链反应检测人树突状细胞 (DCS) 的免疫表型和细胞因子表达。这些结果表明，gRNA/cas9 处理上调了 CD80、CD83 和 CD86 的表达，并显著增加了 IL-6、IL-12、IL-23 和肿瘤坏死因子-α的 mRNA 水平。抗 HBV 和抗 PD-1 联合治疗可抑制 HBV 表达，显著改善 HBV 转基因小鼠的存活率。此外，联合治疗增加了 T 细胞产生的干扰素，进而增强了 Th1 相关免疫刺激基因的表达，从而降低了调节/抑制免疫基因的转录。总的来说，这种反应可以使肿瘤微环境从免疫抑制转变为免疫刺激。最后，抗 HBV 治疗可诱导 HBV 转基因小鼠体内干扰素依赖性程序性细胞死亡配体-1 的表达。总之，这些结果表明，HBV 靶向治疗与 PD-1 免疫检查点阻断联合具有很强的协同作用，从而支持这种联合治疗策略在 HBV 感染临床治疗中的转化潜力。
