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The dynamic changes of circulating invariant natural killer T cells during chronic hepatitis B virus infection

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Abstract

Aim

The protective role of invariant natural killer T cells (iNKTs) against hepatitis B virus (HBV) infection remains controversial. We sought to clarify the role of peripheral iNKT cells during chronic HBV infection.

Methods

Sixty patients with chronic HBV infection were categorized into an immune tolerance phase (HBV-IT) (n = 16), an immune clearance phase (HBV-IC) (n = 19) and an inactive carrier phase (HBV-IA) (n = 25). Twenty healthy individuals were enrolled as healthy controls. Another 21 HBeAg-positive patients were administrated with entecavir (0.5 mg/day) for 6 months. The percentages of circulating iNKT cells and their IFN-γ and IL-4 expression levels were examined by flow cytometry. The relationships between serum HBV DNA, ALT levels, the percentages of iNKT cells, and their IFN-γ and IL-4 levels were analyzed.

Results

Compared to healthy controls, the percentage of iNKT cells decreased in HBV-IT, but increased in HBV-IC and HBV-IA. Circulating IFN-γ-producing iNKT cells gradually increased, whereas IL-4-producing iNKT cells gradually decreased from HBV-IT stage to HBV-IC and HBV-IA stages. The frequency of iNKT cells and their IFN-γ levels were reversely correlated with viral load. The levels of IL-4 expressed by iNKT cells were positively correlated to viral load and the serum ALT levels. After anti-virus therapy, the percentage of IFN-γ-producing iNKT cells increased while the percentage of IL-4-producing iNKT cells decreased.

Conclusions

During chronic HBV infection, the percentages of peripheral iNKT cells and its cytokines expressions of IFN-γ and IL-4 showed dynamic changes. The expression levels of IFN-γ and IL-4 were correlated with the clearance of HBV and liver injury.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China [81202662 and 81473477], and Science Research Project of Twelve Five-year Plan [2012ZX10005004-002], and Doctoral Fund of Ministry of Education of China [20123107110003], and Construction project of TCM clinical research base of State Administration of TCM China [JDZX2012058], and Shanghai Rising-Star Program [13QA1403500], and Three-year action plan of development of TCM in Shanghai [ZYSNXD-CC-ZDYJ015 and ZY3-CCCX-3-3027], and Training plan of outstanding young medical talents, Shanghai Municipal Health Bureau [XYQ2013093].

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    Authors and Affiliations

    1. Laboratory of Cellular Immunity, Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shuguang Hospital, Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, People’s Republic of China

      Man Li, Zhen-Hua Zhou, Xin Zhang, Shu-Gen Jin, Ya-Ting Gao & Yue-Qiu Gao

    2. Department of Hepatopathy, Shuguang Hospital, Affiliated to Shanghai University of Traditional Chinese Medicine, 528 Zhangheng Road, Shanghai, 201203, People’s Republic of China

      Xue-Hua Sun, Xiao-Jun Zhu, Yun Jiang & Yue-Qiu Gao

    3. Department of Infectious Diseases, Changhai Hospital, Second Military Medical University, Shanghai, 200433, People’s Republic of China

      Cheng-Zhong Li

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    1. Man Li
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    Correspondence to Cheng-Zhong Li or Yue-Qiu Gao.

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    All authors concur with the submission and the authors have no potential conflicting interests.

    Ethical standard

    All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2008 (5). This study was approved by the local ethics committee, and all patients provided their written informed consent according to a protocol reviewed and approved by the Institutional Review Board of Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine. All of these patients and normal controls were Chinese. Our study was approved by the local ethics committee, and all patients provided their written informed consent according to a protocol reviewed and approved by the Institutional Review Board of Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine.

    Informed consent

    Informed consent was obtained from all patients for being included in the study.

    Additional information

    Zhen-Hua Zhou has contributed equally to this work.

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    Supplementary material 1 (TIFF 27 kb)

    12072_2015_9650_MOESM2_ESM.tif

    Figure 1 Dynamic changes of peripheral iNKTs and its cytokines expressions during chronic HBV infection. Circulating IFN-γ-producing iNKT cells gradually increased, whereas IL-4-producing iNKT cells gradually decreased from HBV- IT stage to HBV- IC and HBV -IA stages. The frequency of iNKT cells and their IFN-γ levels were reversely correlated to viral load. The levels of IL-4 expressed by iNKT cells were positively correlated to viral load and the serum ALT levels. Supplementary material 2 (TIFF 28 kb)

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    Li, M., Zhou, ZH., Sun, XH. et al. The dynamic changes of circulating invariant natural killer T cells during chronic hepatitis B virus infection. Hepatol Int 10, 594–601 (2016). https://doi.org/10.1007/s12072-015-9650-0

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