Sarkar Neelakshi, Chakravarty Runu, Ganguli Sayak, Singh Shivram Prasad, Narayan Jimmy, Banerjee Arup
ICMR Virus Unit, Kolkata, ID & BG Hospital Campus, Kolkata, India.
Department of Biotechnology, St. Xavier's College (Autonomous), Kolkata, India.
Gastroenterol Hepatol Bed Bench. 2022;15(4):377-386. doi: 10.22037/ghfbb.v15i4.2587.
This study is an attempt to screen the key immune elements that participate during HBV infection and the related pathways that are modulated.
The pathogenesis of Hepatitis B virus and the corresponding clinical manifestations in the host are primarily immune-mediated.
This study utilizes a PCR array to screen immune-related genes that are differentially expressed in the presence of the virus in HBV replicating HepG2.2.15 cells as compared to control HepG2 cells. The significantly up-regulated genes were subjected to bioinformatic analysis utilizing GSEA and STRING. Additionally, ELISA was used to corroborate the levels of Alpha 1 antitrypsin (AAT) from patients' sera.
The expressions of 31% of the studied genes were significantly up-regulated (> 2-fold, p<0.05) in HepG2.2.15 cells compared to controls, and this included the SERPINA1, FN1, IL1R2, LBP, LY96, LYZ and PROC genes. When they were clustered based on biological processes, signaling pathways, and disease progression, the genes related to biotic stimulus, complement-coagulation cascades, and fibrosis, respectively, showed the highest (p<0.05) enrichment. Analysis of patients' sera by ELISA revealed that the serum AAT (SERPINA1) levels were significantly higher in asymptomatic carriers and in patients with chronic liver disease than in controls (p<0.05). Moreover, SERPINA1 levels were also positively correlated with the levels of serum ALT (r=0.4495, p<0.05) among HBV infected patients.
The current study highlights the key immune elements and pathways that are modulated during HBV infection and proposes the possible use of AAT as a non-invasive immunological biomarker to follow the progression of liver disease.
本研究旨在筛选参与乙肝病毒（HBV）感染过程的关键免疫元件以及受调控的相关途径。
乙肝病毒的发病机制及在宿主体内相应的临床表现主要由免疫介导。
本研究利用PCR芯片筛选在HBV复制的HepG2.2.15细胞中与对照HepG2细胞相比，在病毒存在时差异表达的免疫相关基因。对显著上调的基因利用基因集富集分析（GSEA）和STRING进行生物信息学分析。此外，采用酶联免疫吸附测定（ELISA）法检测患者血清中α1抗胰蛋白酶（AAT）水平，以进行验证。
与对照相比，31%的研究基因在HepG2.2.15细胞中的表达显著上调（>2倍，p<0.05），这包括丝氨酸蛋白酶抑制剂A1（SERPINA1）、纤连蛋白1（FN1）、白细胞介素1受体2（IL1R2）、脂多糖结合蛋白（LBP）、髓样分化蛋白2（LY96）、溶菌酶（LYZ）和蛋白酶C（PROC）基因。当根据生物学过程、信号通路和疾病进展进行聚类时，分别与生物刺激、补体 - 凝血级联反应和纤维化相关的基因显示出最高的（p<0.05）富集。ELISA分析患者血清显示，无症状携带者和慢性肝病患者血清中的AAT（SERPINA1）水平显著高于对照组（p<0.05）。此外，在HBV感染患者中，SERPINA1水平也与血清谷丙转氨酶（ALT）水平呈正相关（r = 0.4495，p<0.05）。
本研究突出了HBV感染过程中受调控的关键免疫元件和途径，并提出AAT可能作为一种非侵入性免疫生物标志物用于追踪肝病进展。
