Basic Study
Copyright ©The Author(s) 2017. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Dec 28, 2017; 23(48): 8500-8511
Published online Dec 28, 2017. doi: 10.3748/wjg.v23.i48.8500
Influence of TBX21 T-1993C variant on autoimmune hepatitis development by Yin-Yang 1 binding
Wei Sun, Hong-Yan Wu, Song Chen
Wei Sun, Song Chen, Institute of Infectious Diseases, Southwest Hospital, The Third Military Medical University, Chongqing 400038, China
Hong-Yan Wu, Nuclear Medicine Department, the First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China
Author contributions: Sun W and Wu HY contributed equally to this work; Chen S designed the research study and wrote the paper; Sun W collected and analyzed the data; Wu HY performed the research; all authors have read and approved the final version to be published
Supported by National Natural Science Foundation of China, No. 30972595.
Institutional review board statement: All liver biopsy specimens and blood samples from the patients were taken after informed consent and ethical permission was obtained for participation in the study.
Conflict-of-interest statement: We declare that there are no conflicts of interest related to this study.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Song Chen, MD, Chief Doctor, Professor, Institute of Infectious Diseases, Southwest Hospital, Third Military Medical University, No. 30, Gaotanyan Zhengjie, Shapingba District, Chongqing 400038, China. cs196@medmail.com.cn
Telephone: +86-23-65461319 Fax: +86-23-65461319
Received: June 28, 2017
Peer-review started: July 30, 2017
First decision: August 30, 2017
Revised: October 15, 2017
Accepted: November 8, 2017
Article in press: November 8, 2017
Published online: December 28, 2017
Processing time: 152 Days and 8.5 Hours
ARTICLE HIGHLIGHTS
Background background

Autoimmune hepatitis (AIH) is an autoimmune disease that involves aberrant B and T lymphocyte responses. T-box transcription factor (T-bet) is a key regulator for the lineage commitment in CD4+ T hepler 1 and B effector 1 cells by activating the hallmark production of interferon-γ (IFN-γ). Although the role of T-bet has been studied in a variety of autoimmune diseases in mice, its relative importance in human AIH is not characterized. Detailed knowledge about T-bet-medicated immune responses will therefore enhance our understanding of the pathogenesis of AIH and might support the development of new immunomodulatory treatment approaches.

Research motivation

TBX21, which encodes T-bet, harbors many common polymorphisms at a strong linkage disequilibrium, and distinct TBX21 haplotypes are associated with autoimmune diseases in ethnically distinct populations. Our case-control study (84 cases and 318 control subjects) demonstrated a commone single nucleotide polymorphism (SNP) at the -1993 site of the TBX21 gene promoter that was associated with susceptibility to type 1 AIH (AIH-1) in a chinese population. Individuals carrying the -1993C allele had a decreased risk to AIH-1 compared with those without the -1993C allele (OR = 0.22; 95%CI: 0.09-0.56; P = 0.0016). Functional studies are necessary to dissect the mechanisms underlying the contribution of natural genetic variations to TBX21 dysregulation and the susceptibility to AIH-1 development.

Research objectives

In this study, the authors determined the characterization of transcription factor binding to the T-1993C SNP site of the TBX21 promoter in CD4+ T and B cell lines, and measured the expression of T-bet and IFN-γ in peripheral blood CD4+ T cells and B cells from active AIH-1 patients carrying -1993TC and -1993TT genotypes, compared with healthy controls carrying -1993TC and -1993TT genotypes, in an attempt to provide functional evidence for the association of the TBX21 T-1993C promoter polymorphism and AIH-1 development.

Research methods

In vivo, in vitro, and reporter analyses were performed to determine the function of transcription factor Yin-Yang 1(YY1) binding to the T-1993C element of the TBX21 promoter in human CD4+ T and B cell lines. Flow cytometry and quantitative real-time PCR were used to analyze T-bet and IFN-γ expressions in CD4+ T cells, B cells and monocytes from the peripheral blood of AIH-1 patients including 5-1993TC and 15-1993TT genotype carriers, and healthy controls including 10-1993TC and 25-1993TT genotype carriers. Furthermore, knockdown of YY1 with siRNA was performed to investigate T-bet-medicated regulation of immune response in peripheral blood of AIH-1 patients. The difference between the groups was analyzed by parametric Student’s t-test or nonparametric Mann-Whitney U test. Correlations of T-bet expression in CD4+ T cells and B cells from active AIH-1 patients with AIH disease activity, represented by the International Autoimmune Hepatitis Group score, were examined by Pearson correlation.

Research results

TBX21-1993C allele created a strong Yin-Yang 1 (YY1)-binding site and decreased TBX21 promoter activity in human CD4+ T and B cells. Higher levels of T-bet and IFN-γ were detected in the circulating CD4+ T cells and B cells of AIH-1 patients carrying the TBX21-1993 TT genotype compared with the patients carrying the -1993 TC genotype and controls with the -1993 TC genotype. T-bet expression levels of circulating T cells and B cells were also positively correlated with AIH-1 disease activity. Knockdown of YY1 with siRNA caused increased expression of T-bet and IFN-γ in peripheral blood mononuclear cells in AIH-1 patients. This study has limitations, such as a relative small sample size and the lack of functional study of T-bet-expressing T and B cells in the pathogenic process of AIH, as well as no information about T-bet-expressing T and B cells infiltrates in the liver.

Research conclusions

Reduced T-bet and IFN-γ expression of circulating CD4+ T cells and B cells existed in the individuals carrying the -1993C allele compared with those without the -1993C allele, and played a protective role in AIH-1 development. Moreover, the authors propose a schematic model showing high-affinity binding of YY1 to the -1993C allele site leading to down-regulation of the TBX21 promoter activity, a mechanism for a promoter polymorphism affecting T-bet expression in AIH.

Research perspectives

Further studies are necessary to identify polymorphisms in other loci of the genome and to analyze their association with the T-1993C polymorphism for further elucidation of the genomic background of AIH. Furthermore, further longitudinal studies are necessary to analyze the numbers and function of T-bet-expressing T and B cells in the pathogenic process of AIH -1 with a bigger population.