Expression and functionality of Toll- and RIG-like receptors in HepaRG cells

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• 作者：Souphalone Luangsay¹ ; ² ; ⁵ ; ^&dagger ; ; Malika Ait-Goughoulte¹ ; ² ; ⁵ ; ^&dagger ; ; Maud Michelet¹ ; ² ; Océ ; ane Floriot¹ ; ² ; Marc Bonnin¹ ; ² ; Marion Gruffaz¹ ; ² ; Michel Rivoire³ ; ⁴ ; Simon Fletcher⁵ ; ^§ ; ; Hassan Javanbakht⁵ ; Julie Lucifora¹ ; ² ; Fabien Zoulim¹ ; ² ; ⁶ ; ⁷ ; ^&Dagger ; ; ^{fabien.zoulim@inserm.fr" class="auth_mail" title="E-mail the corresponding author} ; David Durantel¹ ; ² ; ^&Dagger ; ; ^{david.durantel@inserm.fr" class="auth_mail" title="E-mail the corresponding author}
• 关键词：PHH ; primary human hepatocyte ; TLR ; toll-like receptor ; RIG-I ; retinoic acid-inducible gene 1 ; RLR ; RIG-like receptor ; RT ; reverse transcription ; qPCR ; quantitative polymerase chain reaction ; FACS ; fluorescence-activated cell sorting ; IL-6 ; interleukin-6 ; IP-10 ; interferon-gamma-inducible protein-10 ; IFN ; interferon ; HBV ; hepatitis B virus ; MDA-5 ; melanoma differentiation-associated gene 5 ; HAV ; hepatitis A virus ; HEV ; hepatitis E virus ; HDV ; hepatitis D virus ; HCV ; hepatitis C ; WHO ; world healt
• 刊名：Journal of Hepatology
• 出版年：2015
• 出版时间：November 2015
• 年：2015
• 卷：63
• 期：5
• 页码：1077-1085
• 全文大小：1236 K

文摘

HepaRG cells are considered as the best surrogate model to primary human hepatocyte (PHH) culture to investigate host-pathogen interactions. Yet their innate immune functions remain unknown. In this study, we explored the expression and functionality of Toll-like (TLR) and retinoic acid-inducible gene-1 (RIG-I)-like receptors (RLR) in these cells.

Methods

Gene and protein expression levels of TLR-1 to 9 and RLR in HepaRG were mainly compared to PHH, by RT-qPCR, FACS, and Western blotting. Their functionality was assessed, by measuring the induction of toll/rig-like themselves and several target innate gene expressions, as well as the secretion of IL-6, IP-10, and type I interferon (IFN), upon agonist stimulation. Their functionality was also shown by measuring the antiviral activity of some TLR/RLR agonists against hepatitis B virus (HBV) infection.

Results

The basal gene and protein expression profile of TLR/RLR in HepaRG cells was similar to PHH. Most receptors, except for TLR-7 and 9, were expressed as proteins and functionally active as shown by the induction of some innate genes, as well as by the secretion of IL-6 and IP-10, upon agonist stimulation. The highest levels of IL-6 and IP-10 secretion were obtained by TLR-2 and TLR-3 agonist stimulation respectively. The highest preventive anti-HBV activity was obtained following TLR-2, TLR-4 or RIG-I/MDA-5 stimulations, which correlated with their high capacity to produce both cytokines.

Conclusions

Our results indicate that HepaRG cells express a similar pattern of functional TLR/RLR as compared to PHH, thus qualifying HepaRG cells as a surrogate model to study pathogen interactions within a hepatocyte innate system.