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Hepatitis C Virus Entry into Macrophages/Monocytes Mainly Depends on the Phagocytosis of Macrophages

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Abstract

Background

Hepatitis C virus (HCV) has been classified as a strictly hepatotropic pathogen for a long time, and hepatocytes are target cells for HCV infection. More and more studies showed non-liver cells supported HCV entry and replication, such as macrophages. The mechanisms of HCV entry into macrophages are still not clear.

Aims

This study aims to determine the way of HCV entry into macrophages.

Methods

Cell culture-derived infectious HCV particles (HCVcc) were prepared using Huh7 cells transfected with HCV RNA. CD81-knockdown cells were obtained through siRNA transfection. HCV RNA levels were determined by RT-qPCR. Flow cytometry analyses were used to determine cell surface levels of CD11b, CD68, and CD81. ELISA and western blotting were performed to quantify the protein levels of IL-1β, IL-6, and TNF-α. Phagocytic ability was determined by neutral red uptake assay.

Results

CD81 knockdown could not inhibit HCVcc entry into macrophages. The entry of HCV into macrophages could not be blocked by pooled IgG from chronic hepatitis C patient’s sera. Macrophages derived from THP-1 cells displayed stronger phagocytic capacity, which also swallowed more HCV RNA. Treatment of macrophages with endocytic inhibitor, methyl-β-cyclodextrin, decreased the internalization of HCV. HCV uptake by macrophages was related to the reorganization of F-actin cytoskeleton and PI3Ks activation. HCV infection significantly increased the expression of IL1β and IL6 in macrophages and promoted apoptosis of macrophages.

Conclusions

HCV entry into macrophages mainly depends on phagocytosis of macrophages.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (81301445), Applied Basic Research Project of Science and Technology Department in Sichuan (2013JY0173), and China Postdoctoral Science Foundation (2018T111157, 2017M623427, 2017M613427, and 2017T100820).

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Author notes

  1. Yuan Liu and Wenbo Wang have contributed equally to this work.

Authors and Affiliations

  1. Clinical Laboratory, The General Hospital of Western Theater Command, Chengdu, China

    Yuan Liu, Ziying Zou, Zonghai Hu & Jie Xiong

  2. Center for Disease Control and Prevention of Western Theater Command, Chengdu, China

    Wenbo Wang & Quanshui Fan

Authors
  1. Yuan Liu
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  2. Wenbo Wang
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  3. Ziying Zou
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  4. Zonghai Hu
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  6. Jie Xiong
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Contributions

LY and XJ designed the research and analyzed data. LY, WW, ZZ, and XJ wrote the paper. WW, ZZ, HZ, and FQ performed research and analyzed data.

Corresponding authors

Correspondence to Yuan Liu or Jie Xiong.

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Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Ethical statement

This study was conducted according to the principles of the Declaration of Helsinki and approved by the ethics committee of General Hospital of Chengdu Military Region. The use of human blood and sera of chronic hepatitis C patients and animals were received approval (No. CZ1705002). Written informed consent was obtained from all individuals. All samples were anonymized. Protocols for care and use of mice and rabbits are in accordance with Regulations on the Administration of Laboratory Animals, National Science and Technology Commission. Every effort was made to minimize suffering.

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Liu, Y., Wang, W., Zou, Z. et al. Hepatitis C Virus Entry into Macrophages/Monocytes Mainly Depends on the Phagocytosis of Macrophages. Dig Dis Sci 64, 1226–1237 (2019). https://doi.org/10.1007/s10620-018-5401-0

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  • DOI: https://doi.org/10.1007/s10620-018-5401-0

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