Andreas Betz, Hao-En Huang, Zuguang Gu, Ombretta Colasanti, Teng-Feng Li, Jasper Hesebeck-Brinckmann, Nadine Gillich, Gnimah Eva Gnouamozi, Matthias Schlesner, Florian W. R. Vondran, Stephan Urban, Ralf Bartenschlager, Marco Binder, Volker Lohmann
- Abundance of essential components of double-stranded RNA (dsRNA) recognition and the subsequent interferon (IFN) response vary widely between primary human hepatocytes (PHHs) and commonly used cell culture models based on derivatives of Huh7 cells, supporting replication of all hepatitis viruses. We used RNA sequencing to compare the innate immune response in hepatoma cells with primary cells and non-neoplastic immortalized hepatocytes (PH5CH). Stimulation with the dsRNA analog poly(I:C) in Huh7 and Huh7.5, either by supernatant feeding or by transfection, resulted in an induction of interferon-stimulated genes widely comparable to that of PHH and PH5CH, but Huh7 and Huh7.5 lacked efficient production of IFN-β. We identified interferon regulatory factor 7 (IRF7) as a critical component missing in Huh7-derived cells. Upon reconstitution of IRF7 expression, IFNB induction was restored to the levels observed in PHH upon stimulation with poly(I:C). In contrast to PH5CH cells, for whichAbundance of essential components of double-stranded RNA (dsRNA) recognition and the subsequent interferon (IFN) response vary widely between primary human hepatocytes (PHHs) and commonly used cell culture models based on derivatives of Huh7 cells, supporting replication of all hepatitis viruses. We used RNA sequencing to compare the innate immune response in hepatoma cells with primary cells and non-neoplastic immortalized hepatocytes (PH5CH). Stimulation with the dsRNA analog poly(I:C) in Huh7 and Huh7.5, either by supernatant feeding or by transfection, resulted in an induction of interferon-stimulated genes widely comparable to that of PHH and PH5CH, but Huh7 and Huh7.5 lacked efficient production of IFN-β. We identified interferon regulatory factor 7 (IRF7) as a critical component missing in Huh7-derived cells. Upon reconstitution of IRF7 expression, IFNB induction was restored to the levels observed in PHH upon stimulation with poly(I:C). In contrast to PH5CH cells, for which IRF3 was sufficient for full IFN induction, the lack of IRF7 could not be compensated by increased IRF3 expression in Huh7-derived cells. We further found significant IFNB induction upon Sendai virus and hepatitis delta virus infections in Huh7.5 cells reconstituted with IRF7, but not in hepatitis A or hepatitis C virus-infected cells, widely representing the characteristics of the IFN response observed in other models and in vivo. Our data suggest that the reconstitution of IRF7 expression in Huh7 cells can aid a more physiological analysis of cell intrinsic immune responses to hepatotropic viruses in future studies.…
