Original Article
Copyright ©2014 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 14, 2014; 20(26): 8558-8571
Published online Jul 14, 2014. doi: 10.3748/wjg.v20.i26.8558
Changes in circulating Foxp3+ regulatory T cells and interleukin-17-producing T helper cells during HBV-related acute-on-chronic liver failure
Xue-Song Liang, Cheng-Zhong Li, Yin Zhou, Wei Yin, Ya-Yun Liu, Wen-Han Fan
Xue-Song Liang, Cheng-Zhong Li, Yin Zhou, Wei Yin, Ya-Yun Liu, Wen-Han Fan, Department of Infectious Diseases, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
Yin Zhou, Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China
Author contributions: Liang XS conceived the study, participated in the design of the study, performed the statistical analysis and drafted the manuscript; Li CZ participated in the design of the study and enrolled the subjects; Zhou Y performed most of the experiments; Yin W collected all of the human material; Liu YY and Fan WH were responsible for patient follow up; all authors read and approved the final manuscript.
Supported by Grants from Shanghai Natural Science Fund, No. 09ZR1400500; National Natural Science Foundation of China No. 30972600; and Shanghai Health Bureau Fund, No. 2012092
Correspondence to: Xue-Song Liang, Associate Professor, Department of Infectious Diseases, Changhai Hospital, Second Military Medical University, No. 168 Changhai Road, Yangpu District, Shanghai 200433, China. liangxuesong2000@163.com
Telephone: +86-21-31161902 Fax: +86-21-31161910
Received: October 15, 2013
Revised: December 31, 2013
Accepted: April 8, 2014
Published online: July 14, 2014
Processing time: 272 Days and 15.8 Hours
Abstract

AIM: To longitudinally investigate cytokine gene expression and protein levels in Th17 and Treg cells, to observe T-cell phenotypes during hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACHBLF) and to analyze changes in Th17 and Treg phenotypes during disease progression.

METHODS: We measured the expression of seven Th17/Treg differentiation-related genes and serum concentrations of the corresponding cytokines in 18 ACHBLF, 18 chronic hepatitis B (CHB) disease controls and 10 healthy controls (HCs) by real-time quantitative PCR and enzyme linked immunosorbent assay. Peripheral Th17 and Treg cell frequencies were analyzed by flow cytometry.

RESULTS: From the onset of ACHBLF, patients presented with a conductive Th17 differentiation cytokine environment accompanied by high Th17 frequency and high serum IL-17 levels, which were sustained throughout the disease course. The Treg-related cytokine IL-2 and Foxp3 were also up-regulated from disease onset, and Foxp3 gene expression showed a gradually increasing trend during ACHBLF. The circular phenotype of Treg and Th17 cells showed changes from the onset of ACHGLF. At disease onset, Th17 frequency increased significantly compared with both CHB and HCs, but Treg cell frequency decreased significantly compared with CHB. During the ACHBLF event, Th17 frequency remained higher compared with HCs, but decreased sharply from the peak point to the recovery point; Treg cell frequency increased gradually during the ACHBLF event. Treg and Th17 cell counts correlated with ACHBLF development; in all patients, serum IL-17 levels significantly correlated with patient serum ALT levels. In survivors, Th17 frequency at the onset point and the Treg to Th17 ratio at the peak point correlated with the patient’s model for end stage liver disease (MELD) plus sodium (MELD-Na) score. The Treg to Th17 ratio and the Th17 frequency at onset were significant predictors of patient survival. Low Treg/Th17 cell ratios at the onset predicted poor survival. Survivors exhibited an initial decrease in the circulating Treg/Th17 ratio from the onset to the peak time, and subsequently displayed a continuous increase.

CONCLUSION: Treg and Th17 cells showed changes in genes, protein levels and T cell phenotypes during ACHBLF events. An increased Treg/Th17 ratio was associated with the survival of ACHBLF patients.

Keywords: Hepatitis B virus, Treg, Th17, Immune, Hepatitis B virus-related acute-on-chronic liver failure

Core tip: In this study, we longitudinally investigated Foxp3+Treg cells and IL-17+Th cells by measuring gene levels, protein levels and T-cell phenotypes during HBV-related acute-on-chronic liver failure (ACHBLF) progression. From the onset of ACHBLF, there were changes in Foxp3+Treg and Th17 cell frequencies. An increased Treg/Th17 ratio was associated with the survival of ACHBLF patients.