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Influences of DMSO and Dexamethasone on HBV Infection in HepCHLine-4 Cells

Author: GuanYanYan
Tutor: MaLiXian
School: Shandong University
Course: Internal Medicine
Keywords: Hepatitis B virus Dimethyl sulfoxide Dexamethasone Infection
CLC: R512.62
Type: Master's thesis
Year: 2011
Downloads: 3
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Abstract


BackgroundHepatitis B virus (HBV) is a member of the Hepadnavirus family. Viral infection may progress to acute or chronic hepatitis, and may eventually cause cirrhosis and hepatocellular carcinoma (HCC). According to the World Health Organisation, more than 2 billion people alive today have been infected by HBV, and more than 350 million people are chronically infected. Worldwide deaths from chronic liver diseases and hepatocellular carcinoma caused by HBV infection approximately 1 million each year. China is highly endemic areas. It is estimated that nearly 60%the population has been infected by HBV, and 8% to 15% of them become chronically infected. Up to 16.79% in the highest province, far above the world average Infection rate of about 5%.Since HBV surface antigen was found by Blumberg to the complete cloning and sequencing of HBV DNA genome, our knowledge on genetic constitution, biology feature and life cycle of HBV have been riched. Despite considerable advances in the understanding of the natural history of HBV disease, most of the early steps in the virus life cycle remain unclear. Major obstacle to study HBV infection is lack of a suitable animal model and in vitro cell culture system.We have successfully established a new cell model previously, which was a hybrid cell line by fused HepG2 cells with primary human hepatocytes, named HepCHLine-4 cell line. The results indicate that the hybrid cell line HepCHLine-4 inherits the susceptibility to HBV from human hepatocytes; and at the same time, it inherits the immortality of HepG2 cells, overcoming the deficiency of primary human hepatocytes and HepG2 cells as the HBV infection system respectively. HepCHLine-4 cell line is susceptible to HBV and can initiate viral replication, produce HBV-specific antigens and HBV-specific progeny virions after HBV infection. However, the infectability is low relatively. In this study, we successfully improved the susceptibility by adding chemical substances in the medium, which making the hybrid cell line HepCHLine-4 can be the better as a suitable cell model for studying the early events of HBV infection.ObjectiveTo evaluate the influence of dimethyl sulfoxide (DMSO) and dexamethasone to the ability of HepCHLine-4 cells to Hepatitis B virus (HBV) natural infection.MethodsHepCHLine-4 cells were devided into four groups:DMSO group, HepCHLine-4 cells were cultured in Dulbecco s modified Eagle s medium(DMEM) with 20mL/L DMSO for 4 days; dexamethasone group, HepCHLine-4 cells were cultured in DMEM with 5×10-5mol/L dexamethasone for 4 days; DMSO&dexamethasone group, HepCHLine-4 cells were cultured in DMEM with 20mL/L DMSO and 5x10-5mol/L dexamethasone for 4 days; and control group, HepCHLine-4 cells were cultured in DMEM without DMSO and dexamethasone for 4 days. Then HepCHLine-4 cells were incubated with HBV for 24 hours at 37℃. Hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in the supernatants of infected cells were detected by electrochemiluminescence(ECL). HBV DNA in the supernatants of infected cells was detected by fluorescence quantitative real-time polymerase chain reaction (FQ-RCR). Hepatitis B core antigen (HBcAg) in HBV-infected cells was detected by indirect immunofluorescence assay.ResultsDMSO group:the titer of HBsAg and HBeAg is relatively high; dexamethasone group:the titer of HBV DNA is relatively high; DMSO&dexamethasone group:the titer of HBsAg and HBeAg and the titer of HBV DNA are higher, the highest levels are 125.790IU/mL,4.784S/Co and 5.930x105 copies/mL respectively; control group: the titer of HBsAg and HBeAg and the titer of HBV DNA are lower, the highest levels are 85.490IU/mL,1.896S/Co,3.729×104copies/mL respectively. HBcAg was detected throughout the cytoplasm of HBV-infected cells, however, among the four groups, pigmentation in the DMSO & Dexamethasone group was more evident.ConclusionDMSO and dexamethasone can improve the ability of HepCHLine-4 to HBV natural infection.

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CLC: > Medicine, health > Internal Medicine > Infectious disease > Viral infections > Viral Hepatitis > Hepatitis B
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