Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, China.
Front Immunol. 2023 Feb 9;14:1121795. doi: 10.3389/fimmu.2023.1121795. eCollection 2023.
Chronic hepatitis B (CHB) virus infection is a major risk factor for cirrhosis and hepatocellular carcinoma (HCC). Hepatitis B virus (HBV) immune escape is regulated by the exhaustion of virus-specific CD8 T cells, which is associated with abnormal expression of negative regulatory molecule CD244. However, the underlying mechanisms are unclear. To investigate the important roles of non-coding RNAs play in CD244 regulating HBV immune escape, we performed microarray analysis to determine the differential expression profiles of long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and mRNAs in patients with CHB and patients with spontaneous clearance of HBV. Competing endogenous RNA (ceRNA) was analyzed by bioinformatics methods and confirmed by the dual-luciferase reporter assay. Furthermore, gene silencing and overexpression experiments were used to further identify the roles of lncRNA and miRNA in HBV immune escape through CD244 regulation. The results showed that the expression of CD244 on the surface of CD8 T cells was significantly increased in CHB patients and in the co-culture system of T cells and HBV-infected HepAD38 cells, which was accompanied by the reduction of miR-330-3p and the elevation of lnc-AIFM2-1. The down-regulated miR-330-3p induced the apoptosis of T cells by lifting the inhibition of CD244, which was reversed by miR-330-3p mimic or CD244-siRNA. Lnc-AIFM2-1 promotes the accumulation of CD244, which is mediated by decreased miR-330-3p, and then reduced the clearance ability of CD8 T cells to HBV through regulated CD244 expression. And the injury in the ability of CD8 T cells to clear HBV can be reversed by lnc-AIFM2-1-siRNA, miR-330-3p mimic, or CD244-siRNA. Collectively, our findings indicate that lnc-AIFM2-1 on CD244 by acting as a ceRNA of miR-330-3p contributes to HBV immune escape, which may provide novel insights into the roles of interaction networks among lncRNA, miRNA, and mRNA in HBV immune escape, highlighting potential applications of lnc-AIFM2-1 and CD244 for diagnosis and treatment in CHB.
慢性乙型肝炎（CHB）病毒感染是肝硬化和肝细胞癌（HCC）的主要危险因素。乙型肝炎病毒（HBV）免疫逃逸受病毒特异性 CD8 T 细胞耗竭的调节，这与负调节分子 CD244 的异常表达有关。然而，其潜在机制尚不清楚。为了研究非编码 RNA 在 CD244 调节 HBV 免疫逃逸中的重要作用，我们进行了微阵列分析，以确定 CHB 患者和 HBV 自发清除患者中长链非编码 RNA（lncRNA）、microRNA（miRNA）和 mRNA 的差异表达谱。通过生物信息学方法分析竞争性内源性 RNA（ceRNA），并通过双荧光素酶报告基因检测进行验证。此外，通过基因沉默和过表达实验，进一步通过 CD244 调节鉴定 lncRNA 和 miRNA 在 HBV 免疫逃逸中的作用。结果表明，CHB 患者和 T 细胞与 HBV 感染的 HepAD38 细胞共培养系统中，CD8 T 细胞表面 CD244 的表达明显增加，同时 miR-330-3p 下调，lnc-AIFM2-1 上调。下调的 miR-330-3p 通过解除对 CD244 的抑制作用诱导 T 细胞凋亡，该作用可被 miR-330-3p 模拟物或 CD244-siRNA 逆转。lnc-AIFM2-1 通过降低 miR-330-3p 促进 CD244 的积累，然后通过调节 CD244 表达降低 CD8 T 细胞对 HBV 的清除能力。通过 lnc-AIFM2-1-siRNA、miR-330-3p 模拟物或 CD244-siRNA 可逆转 CD8 T 细胞清除 HBV 的能力损伤。总之，我们的研究结果表明，lnc-AIFM2-1 通过作为 miR-330-3p 的 ceRNA 作用于 CD244，有助于 HBV 免疫逃逸，这可能为 lncRNA、miRNA 和 mRNA 相互作用网络在 HBV 免疫逃逸中的作用提供新的见解，强调了 lnc-AIFM2-1 和 CD244 在 CHB 诊断和治疗中的潜在应用。
