5-Aza-2’-deoxycytidine may enhance the frequency of T regulatory cells from CD4⁺ naïve T cells isolated from the peripheral blood of patients with chronic HBV infection

ABSTRACT

Objectives

Methylation pattern of gene modification is essential for the differentiation of T regulatory cells (Tregs) and 5-Aza-2ʹ-deoxycytidine is a common inhibitor of methylation. This study aimed to investigate the potential effects of Treg polarizing conditions and 5-Aza-2ʹ-deoxycytidine treatment in the differentiation of naïve T cells during chronic hepatitis B virus (HBV) infection.

Methods

The frequency of Tregs in peripheral blood was determined by flow cytometry from patients with chronic hepatitis B (CHB) (n = 51), liver cirrhosis (LC) (n = 47), hepatocellular carcinoma (HCC) (n = 40) and healthy controls (HCs) (n = 17). Gene expression were detected by qRT-PCR and DNA methyltransferases (DNMT) Activity was also determined.

Results

The frequency of Tregs and Foxp3 expression in peripheral blood from 5-Aza-2ʹ-deoxycytidine-treated groups were higher than that with acetic acid treatment as a control. Foxp3 mRNA and the frequency of Tregs derived from naïve CD4⁺T cells from peripheral blood of patients with HCC or LC were more pronounced compared with HCs. 5-Aza-2ʹ-deoxycytidine may have induced a more pronounced upward trend of PD-1 expression in HBV patients.

Conclusions

5-Aza-2ʹ-deoxycytidine mediated demethylation has potential effects on enhancing the differentiation of naïve T cells to Tregs in chronic HBV infection.

KEYWORDS:

• Hepatitis B virus
• methylation
• 5-Aza-2ʹ-deoxycytidine
• t regulatory cells
• pd-1
• dna methyltransferases

Declaration of interest

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in this manuscript.

Reviewer disclosures

Peer reviewers on this manuscript have no relevant financial or other relationships to disclose.

Author contributions

All authors have made contributions to the study and approved the publication. Yu Fang, Xiao-Dong Yuan and Hui-Hui Liu contributed to the majority of the experiments and data collection. Lin Xiang and La-Mei Chen and Shuai Gao completed statistical analysis and interpretation of data. Yu-Chen Fan and Kai Wang and contributed to the study designed and drafted the manuscript. Yu-Chen Fan and Yu Fang contributed to revised the manuscript. Kai Wang was responsible for study supervision and obtaining funding.

Additional information

Funding

This work was supported by the Ministry of Science and Technology of the People's Republic of China (2017ZX10202202 and 2018ZX10302206), the National Natural Science Foundation of China (81970522), the Key Research and Development Project of Shandong Province (2019GSF108023), China Postdoctoral Science Foundation（2020M672074), and Shandong University multidisciplinary research and innovation team of young scholars (2020QNQT11).