Novel entry pathways for Hepatitis B virusTools Parmar, Devan (2023) Novel entry pathways for Hepatitis B virus. MRes thesis, University of Nottingham.
AbstractHepatitis B virus (HBV) is a global health problem. Patients with chronic hepatitis B (CHB) infection can develop progressive liver disease, including cirrhosis and hepatocellular carcinoma (HCC). There is still much to understand about the molecular biology of HBV, particularly viral entry. The main entry receptor of HBV is sodium taurocholate cotransporting polypeptide (NTCP); however, a variant of HBV, BT10D4 has recently been discovered, that infects cells independent of NTCP expression. This study aimed to demonstrate NTCP-independent infection by BT10D4 and to elucidate the alternative receptor involved in entry of this variant. The methodology used to investigate the entry receptor is known as pseudotyping, a technique utilised to produce ‘customized’ chimeric viruses that possess envelope glycoproteins that are not encoded by the viral genome. In this case, the pseudotype has an HBV glycoprotein and a HIV-1 capsid containing a luciferase reporter gene, allowing quantification of infection into susceptible cells. Pseudotypes were used to infect the hepatoma cell line Huh7, and Huh7 cells induced to express NTCP. The pseudotype assays clearly demonstrated that BT10D4 facilitated NTCP-independent infection. Furthermore, the receptor annexin A2 was identified as a candidate for
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