在hbv阳性人肝细胞癌细胞系中基于咪喹莫特的toll样受体7配体

StephenW

BMC Infect Dis. 2017 Jan 14;17(1):76. doi: 10.1186/s12879-017-2189-z.
Anti-hepatitis B virus (HBV) response of imiquimod based toll like receptor 7 ligand in hbv-positive human hepatocelluar carcinoma cell line.Das D1, Sengupta I2, Sarkar N1, Pal A1, Saha D1, Bandopadhyay M1, Das C2, Narayan J3, Singh SP3,4, Chakrabarti S1,5, Chakravarty R6.
Author information

• 1ICMR Virus Unit, Kolkata, ID & BG Hospital Campus, ICMR Virus Unit, GB 4, 700010, Kolkata, India.
• 2Biophysics & Structural Genomics Division, Saha Institute of Nuclear Physics, Kolkata, India.
• 3Department of Gastroenterology, SCB Medical College, Cuttack, India.
• 4Kalinga Gastroenterology Foundation, Beam Diagnostics Premises, Cuttack, India.
• 5National Institute of Cholera and Enteric Diseases, Kolkata, India.
• 6ICMR Virus Unit, Kolkata, ID & BG Hospital Campus, ICMR Virus Unit, GB 4, 700010, Kolkata, India. [email protected].
  

AbstractBACKGROUND: Toll like receptors (TLRs) play an important role in innate immunity and various studies suggest that TLRs play a crucial role in pathogenesis of hepatitis B virus (HBV) infection. The present study aims in looking into the status of crucial host and viral gene expression on inciting TLR7.
METHODS: The transcription of TLR7 pathway signaling molecules and HBV DNA viral load were quantified by Real Time-PCR after stimulation of TLR7 with its imiquimod based ligand, R837. Cell cycle analysis was performed using flow-cytometry. Expression of TLR7 and chief cell cycle regulator governing G1/S transition, p53 was also seen in liver biopsysss samples of CHB patients. HBV induced alteration in histone modifications in HepG2 cells and its restoration on TLR7 activation was determined using western blot.
RESULTS: The TLR7 expression remains downregulated in HepG2.2.15 cells and in liver biopsy samples from CHB patients. Interestingly HBV DNA viral load showed an inverse relationship with the TLR7 expression in the biopsy samples. We also evaluated the anti-viral activity of R837, an agonist of TLR7. It was observed that there was a suppression of HBV replication and viral protein production upon TLR7 stimulation. R837 triggers the anti-viral action probably through the Jun N-terminal Kinase (JNK) pathway. We also observed a downregulation of histone H3K9Me3 repression mark upon R837 treatment in HBV replicating HepG2.2.15 cells, mimicking that of un-infected HepG2 cells. Additionally, the G1/S cell cycle arrest introduced by HBV in HepG2.2.15 cells was released upon ligand treatment.
CONCLUSION: The study thus holds a close insight into the changes in hepatocyte micro-environment on TLR7 stimulation in HBV infection.


KEYWORDS: Cell-cycle arrest; Epigenetics; Hepatitis B virus; Innate immune response; TLR7

PMID:28088184DOI:10.1186/s12879-017-2189-z

StephenW

BMC Infect Dis。 2017 Jan 14; 17（1）：76。 doi：10.1186 / s12879-017-2189-z。
在hbv阳性人肝细胞癌细胞系中基于咪喹莫特的toll样受体7配体的抗乙型肝炎病毒（HBV）应答。
Das D1，Sengupta I2，Sarkar N1，Pal A1，Saha D1，Bandopadhyay M1，Das C2，Narayan J3，Singh SP3,4，Chakrabarti S1,5，Chakravarty R6。
作者信息

    1ICMR病毒单位，加尔各答，ID＆BG医院校园，ICMR病毒单位，GB 4,700010，加尔各答，印度。
    2Biophysics＆Structural Genomics Division，Saha Institute of Nuclear Physics，Kolkata，India。
    3D消化科，SCB医学院，Cuttack，印度。
    4Kalinga胃肠病学基金会，Beam Diagnostics前提，Cuttack，印度。
    5国家霍乱和肠道疾病研究所，加尔各答，印度。
    6ICMR病毒单位，加尔各答，ID＆BG医院校园，ICMR病毒单位，GB 4,700010，加尔各答，印度。 [email protected]。

抽象
背景：

Toll样受体（TLR）在先天免疫中起重要作用，并且各种研究表明TLR在乙型肝炎病毒（HBV）感染的发病机理中起关键作用。本研究旨在调查关键宿主和病毒基因表达鞭打TLR7的状态。
方法：

在TLR7以其基于咪喹莫特的配体R837刺激后，通过实时PCR定量TLR7途径信号分子和HBV DNA病毒载量的转录。使用流式细胞术进行细胞周期分析。表达TLR7和主要细胞周期调节器控制G1 / S过渡，p53也见于CHB患者的肝活检样本。 HBV诱导的HepG2细胞中组蛋白修饰的改变及其对TLR7活化的恢复使用蛋白质印迹法测定。
结果：

TLR7表达在HepG2.2.15细胞和来自CHB患者的肝活检样品中保持下调。有趣的是，HBV DNA病毒载量与活检样品中的TLR7表达呈反比关系。我们还评估了R837，TLR7的激动剂的抗病毒活性。观察到在TLR7刺激时存在HBV复制和病毒蛋白质产生的抑制。 R837触发抗病毒作用可能通过Jun N-末端激酶（JNK）途径。我们还HBV复制的HepG2.2.15细胞经R837治疗观察组蛋白的H3K9me3压制商标的下调，模仿，联合国感染的HepG2细胞。此外，通过配体处理释放HBV在HepG2.2.15细胞中引入的G1 / S细胞周期停滞。
结论：

因此，本研究对HBV感染中TLR7刺激的肝细胞微环境的变化有着深入的了解。
关键词：

细胞周期阻滞;表观遗传学;乙型肝炎病毒;先天免疫反应; TLR7

PMID：
    28088184
DOI：
    10.1186 / s12879-017-2189-z

MP4

咪喹莫特是已经上市的药物，不过是外用剂，用来疫苗皮肤激动也许管用