Polarization to Anti-Inflammatory Status of Macrophages Predicts Poor Prognosis of HBV-Related Acute-on-Chronic Liver Failure Patients via Chloride Intercellular Channel 3 Suppression

Background: Patients with HBV-related acute-on-chronic liver failure (HBV-ACLF) are characterized by immune paralysis and susceptibility to sepsis. Macrophages are central mediators of immunoreaction, and reported to polarize to pro-inflammatory or anti-inflammatory functional status in different morbid states. The role of chloride intercellular channel 3 (CLIC3) in immune function is little known.   Aims: To investigate the dynamic changes of macrophage polarization in different stages of HBV-related liver diseases, explore the role of CLIC3 in macrophage polarization and its mechanism. Methods: Peripheral blood mononuclear cells derived macrophages from healthy controls (n=10), mild chronic hepatitis B patients (n=226), HBV-related compensated cirrhosis patients (n=36), HBV-related decompensated liver cirrhosis patients (n=40) and HBV-related acute-on-chronic failure patients (n=62) were collected to analyze the macrophage polarization using flowcytometry. Transcriptome sequencing analysis was performed for better understanding the roles of macrophages in HBV-ACLF. Virto cell experiments were used to investigate the role of CLIC3 in macrophage polarization. Results: The anti-inflammatory, CD163+CD206+ macrophages, significantly increased in HBV-ACLF patients, which predicted a poor clinical outcome. Transcriptome sequencing analysis indicated, CLIC3 was declined in HBV-ACLF patients, negatively related with V-set and immunoglobulin domain containing 4, involving in immunological disease, organismal injury and abnormalities. Virto cell experiments showed CLIC3 regulated macrophage polarization through NF-kB and AKT pathway. And CLIC3 suppression in macrophages promoted anti-inflammatory polarization and aggravated the impair of hepatocytes. Conclusion: Polarization to anti-inflammatory status of macrophages predicts poor prognosis of HBV-ACLF patients. And CLIC3 plays a vital role in macrophage polarization in HBV-ACLF progression.   Funding Statement: This study was supported by the Guangzhou city science and technology project (No. 201607010064), and the Natural Fund of Guangdong Province (No. 2016A030313237 and 2015A030313172). Third Affiliated Hospital of Sun Yat-Sen University Five-Fiver Project (No. k00005). Declaration of Interests: The authors declare no conflict of interest. Ethics Approval Statement: This study was approved by the Third Affiliated Hospital of Sun Yat-Sen University ethics committee ([2018]02-432-01). If patients were unable to provide consent, informed consent was obtained by the next of kin.