Centre for Liver Research, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research (I.P.G.M.E. & R.), Kolkata, India.
Department of Gastroenterology, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research (I.P.G.M.E. & R.), Kolkata, India.
Sci Rep. 2018 May 23;8(1):8055. doi: 10.1038/s41598-018-26414-4.
Distinct clinical features of HBV infection have been associated with different viral genotype/subgenotype. HBV Genotype-D comprised of 10 subgenotypes, D1-D10, whose clinical implications still remain elusive. We investigated for the first-time, the virologic characteristics and cytopathic effects of four non-recombinant D-subgenotypes, D1/D2/D3/D5. Expressions of viral/host genes were evaluated in Huh7 cells transfected with full-length, linear-monomers of HBV/D-subgenotypes or pGL3-Basic vector carrying subgenotype-specific HBx. Intracellular HBV-DNA and pregenomic-RNA levels were high in D1/D2 than D3/D5. Expressions of PreC-mRNA and HBx were highest for D2 and D1 respectively, whereas PreS2/S-transcript was significantly reduced in D5. Increased apoptotic cell death and marked upregulation in caspase-3/Bax/TNF-R1/FasR/TRAIL-R1/ROS/MCP-1/IP-10/MIP-1β expression were noticed specifically in D2- and also in D3-transfected cells, while D5 resulted in over-expression of ER-stress-markers. D-subgenotype-transfected Huh7 cells were co-cultured with PBMC of healthy-donors or LX-2 cells and significant increase in pro-inflammatory cytokines in PBMC and fibrogenic-markers in LX-2 were noticed in presence of D2/D3. Further, Huh7 cells transfected with D1, in particular and also D5, displayed remarkable induction of EMT-markers and high proliferative/migratory abilities. Collectively, our results demonstrated that D2/D3 were more associated with hepatic apoptosis/inflammation/fibrosis and D1/D5 with increased risk of hepatocarcinogenesis and emphasize the need for determining HBV-subgenotype in clinical practice.
HBV 感染的不同临床特征与不同的病毒基因型/亚型有关。HBV 基因型-D 由 10 个亚型（D1-D10）组成，其临床意义仍不清楚。我们首次研究了四个非重组 D 亚基因型（D1/D2/D3/D5）的病毒学特征和细胞病变效应。通过转染全长线性单体 HBV/D 亚基因型或携带亚基因型特异性 HBx 的 pGL3-Basic 载体，在 Huh7 细胞中评估病毒/宿主基因的表达。D1/D2 中的细胞内 HBV-DNA 和前基因组 RNA 水平高于 D3/D5。PreC-mRNA 和 HBx 的表达在 D2 和 D1 中最高，而 D5 中的 PreS2/S-转录物显著减少。在 D2 和 D3 转染的细胞中观察到凋亡细胞死亡增加和 caspase-3/Bax/TNF-R1/FasR/TRAIL-R1/ROS/MCP-1/IP-10/MIP-1β 的表达明显上调，而 D5 导致 ER 应激标志物的过度表达。D 亚基因型转染的 Huh7 细胞与健康供体的 PBMC 或 LX-2 细胞共培养，在 D2/D3 存在的情况下，PBMC 中的促炎细胞因子和 LX-2 中的成纤维标志物显著增加。此外，用 D1，特别是 D5 转染的 Huh7 细胞显示出 EMT 标志物的显著诱导和高增殖/迁移能力。总之，我们的研究结果表明，D2/D3 与肝凋亡/炎症/纤维化的相关性更高，而 D1/D5 与肝癌发生的风险增加有关，这强调了在临床实践中确定 HBV 亚型的必要性。
