A visualizable hepatitis A virus and hepatitis C virus coinfection model in vitro: coexistence of two hepatic viruses under limited competition in viral RNA synthesis

Abstract

Hepatitis A virus (HAV) and hepatitis C virus (HCV) coinfection in patients usually leads to HCV suppression or clearance. Whether this suppression/clearance is caused by direct virus-virus interaction or indirect interactions is still unknown. Here, we present a robust and visualizable HAV/HCV coinfection model for investigating their interactions in vitro. We find that HAV superinfects HCV-persistently-infected Huh-7.5.1 cells without obvious virus-virus exclusion and vice versa, while there is a mild reciprocal viral interference in coinfection. Through single-cell scale confocal microscopy analysis and treating co-infected cells with rNTPs or antivirals e.g. Sofosbuvir, Simeprevir, and Ledipasvir, we find that HAV and HCV exploit cellular machinery in a compatible manner, but with reciprocal competition for rNTPs in their RNA synthesis. In conclusion, our findings reveal the absence of direct HAV-HCV interaction but the presence of indirect interaction, which may be due to limited competition in viral RNA synthesis. Therefore, we propose that suppression/clearance of HCV in HAV/HCV coinfected patients is probably due to indirect interactions e.g. viral competition or immunological interactions. Author summaryVirus-virus interactions could usually be categorized in three aspects: (1) direct interaction induced by interaction of viral gene or gene products; (2) indirect interaction mediated by immune response or (3) host environmental alteration. Hepatitis A virus (HAV) and hepatitis C virus (HCV) are two important causatives of human hepatitis. In patients, their coinfection often induces HCV suppression/clearance, and sometimes may lead to fulminant hepatitis. In this study, we find that a limited reciprocal viral interference occurs in HAV/HCV coinfection in vitro, which results from a viral competition of rNTPs in viral RNA synthesis. Our study provides a new paradigm of studying virus-virus interactions in single-cell scale. To our knowledge, we identify the rNTPs competition in virus coinfection for the first time by experiment, though several mathematical modelings have predicted the resource competition in virus coinfection.

Dual Infection of Hepatitis A Virus and Hepatitis E Virus— What Is Known?

“…HCV infection inhibits HEV replication via HCV protease NS3/4A, which probably cleaves HEV ORF1 to a less active form [ 134 ]. On the other hand, the HAV/HCV dual infection with Huh7.5 showed that there were no direct interactions between the two viruses and limited competition [ 135 ]. Future studies should study the impact of HAV/HEV dual infection in vitro.…”

Modeling Hepatotropic Viral Infections: Cells vs. Animals

“…Their finding revealed that the simultaneous presence of HAV-HCV did not affect the viral RNA synthesis of both viruses. They suggested that indirect interactions may lead to the suppression and clearance of HCV in HAV/HCV coinfected patients [ 55 ]. Sun and colleagues reported the creation of stable TetOFF hepatoma cell lines (HepG2 and Huh7) to control HBV production.…”

“…Primary cells Adult PHH [15,21] Adult PHH [17] Adult PHH [18][19][20][21] Adult PHH [18] primary human intestinal cells [23] primary human endometrial cells [24] Cancer cell lines HepAD38 [34] HepDE19 [35] Hepa RG [44,46] HLCZ01 [36] (HepNB2.7) [38] HrpaRG [39,40] HepG2 +Huh-7 [49,55] HepG2 [51] HepG2-NTCP [52] HLC [24,26,64] HLCZ01 [36] HepG2 [ [150] Abbreviations: ETV, entecavir; FRG, fumaryl acetoacetate hydrolase (Fah)/RAG2/interleukin (IL) 2-gammaC; FRGS, Fah −/− Rag2 −/− IL-2Rγc −/− SCID; HEV, hepatitis E virus; HBV, hepatitis B virus; HCV, hepatitis C virus; IFN, interferon; IG, immunoglobulin; NK, natural killer; NM23TC, nanoformulation of the modified lamivudine 3TC; PRI-724, selective inhibitor of β-catenin/CBP; RG7834, inhibitor of HBV expression; uPA/RAG-2, urokinase-type plasminogen activator/recombinant activation gene-2; uPA/SCID, urokinase-type plasminogen activator/severe combined immunodeficiency; WHBV, woodchuck hepatitis B virus; WMHBV, woolly monkey hepatitis B virus.…”