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Reduction of natural killer cells is associated with poor outcomes in patients with hepatitis B virus-related acute-on-chronic liver failure

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Abstract

Background and aims

Natural killer (NK) cells are critical innate effectors that respond to viral infections and contribute to immunopathology. Here, we aimed to investigate the role of NK cells in hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) and elucidate the underlying mechanism by examining their phenotypic and functional profiles.

Methods

We included patients with HBV-ACLF (n = 37) and chronic hepatitis B (n = 19), and healthy controls (n = 13) in our study. We examined the phenotype and function of different subsets of peripheral NK cells using flow cytometry and RNA-sequencing analysis, and screened liver NK cells using immunohistochemistry. We detected inflammatory cytokines using a Luminex assay. In addition, we analyzed the relationships between these parameters and disease severity.

Results

Peripheral NK cells were decreased and characterized by high expression of caspase-3, Ki67, CXCR3, NKG2D, NKp46, CD107a, and GM-CSF, and typified by higher cell migration and immune response by RNA-sequencing analysis in patients with HBV-ACLF than in those with chronic hepatitis B. Accumulations of CXCL-10 and NK cells were found in the liver, and excessive production of CXCL-10 in the peripheral blood contributed to the apoptosis of NK cells in vitro. The decrease in NK cells was associated with the level of HBV DNA and disease severity and had good prognostic performance in predicting the outcome of patients with HBV-ACLF through AUROC analysis.

Conclusion

NK cells were significantly decreased and showed dysfunction of phenotypic and functional profiles across distinct subsets in the peripheral blood of patients with ACLF. Crosstalk between CXCL-10 and NK cells may mediate the unbalanced distribution of NK cells. Understanding the dysfunction and decrease in NK cells may provide new insights into ACLF pathogenesis.

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Data availability

The datasets generated for this study are available on request to the corresponding authors.

Abbreviations

ACLF:

Acute-on-chronic liver failure

MONO:

Monocytes

NK:

Natural killer

PLT:

Platelet

HBV:

Hepatitis B virus

HBG:

Hemoglobin

IL:

Interleukin

TBil:

Total bilirubin

TNF:

Tumor necrosis factor

ALT:

Alanine aminotransferase

HLA-DR:

Human leukocyte antigen-DR

AST:

Aspartate amino transaminase

HC:

Healthy control

ALB:

Albumin

CHB:

Chronic hepatitis B

sCr:

Serum creatine

APASL:

Asian Pacific for the Study of Liver

MELD:

Model for end-stage liver disease

HCV:

Hepatitis C virus

EDTA:

Ethylene diamine tetraacetic acid

HIV:

Human immunodeficiency virus

PBMC:

Peripheral blood mononuclear cells

PT:

Prothrombin time

G-CSF:

Granulocyte colony-stimulating factor

PTA:

Prothrombin time activity

GB:

Granzyme B

INR:

International normalized ratio

GM-CSF:

Granulocyte–macrophage colony-stimulating factor

WBC:

White blood cell

GO:

Gene ontology

NEUT:

Neutrophils

IFN-γ:

Interferin-γ

LY:

Lymphocytes

AUROC:

Area under the receiver operating characteristic curve

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Acknowledgements

The authors acknowledge the patients, study investigators, and coordinators for their contributions in this study.

Funding

We acknowledge funding support from the National Natural Innovation Fund (Project 81721002) and the National Key R&D Program of China (2017YFA0105703).

Author information

Author notes

  1. Hua-Jie Li and Ning Yang contributed equally to this work.

Authors and Affiliations

  1. Clinical Medical School, Peking University 302, Beijing, 100039, China

    Hua-Jie Li, Xiuying Mu, Ying-Ying Yu, Jing Li & Fu-Sheng Wang

  2. Department of Infectious Diseases, The Fifth Medical Center of Chinese PLA General Hospital, National Clinical Research Center for Infectious Diseases, 100 Xisihuan Road, Fengtai District, Beijing, 100039, China

    Hua-Jie Li, Xiuying Mu, Lili Tang, Song-Shan Wang, Chun-Bao Zhou, Jin-Hong Yuan, Hai-Yan Wang, Ying-Ying Yu, Jing Li, Si-Yuan Chen, Tao Yang, Kai Liu, Wen-Jing Cao, Ming-Ju Zhou, Chao Zhang, Ji-Yuan Zhang, Yan-Mei Jiao, Jin-Wen Song, Xing Fan, Ming Shi, Ruonan Xu & Fu-Sheng Wang

  3. Clinical Testing Center, The Fifth Medical Center of General Hospital of Chinese PLA General Hospital, Beijing, 100039, China

    Ning Yang

  4. The Second School of Clinical Medicine, Southern Medical University, Guangzhou, 510000, China

    Zhi-Qian Feng

  5. The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230000, China

    Hai-Yan Wang & Wen-Jing Cao

  6. Beijing Ditan Hospital, Capital Medical University, Beijing, 100102, China

    Ming-Ju Zhou

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Contributions

H-JL, NY, XM, LT, S-SW, C-BZ, J-HY, H-YW, Y-YY, JL, S-YC, Z-QF, TY, KL, W-JC, M-JZ, CZ, J-YZ, Y-MJ, J-WS, XF and MS participated in the data acquisition and soluble cytokines analysis; H-JL, NY, RX, and F-SW designed the study; H-JL and NY performed analyses and interpretation of data; H-JL and NY wrote the first draft of the manuscript and incorporated revisions; H-JL, NY, RX, and F-SW prepared the final version. All authors approved the final manuscript to be published.

Corresponding authors

Correspondence to Ruonan Xu or Fu-Sheng Wang.

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Conflict of interest

Hua-Jie Li, Ning Yang, Xiuying Mu, Lili Tang, Song-Shan Wang, Chun-Bao Zhou, Jin-Hong Yuan, Hai-Yan Wang, Ying-Ying Yu, Jing Li, Si-Yuan Chen, Zhi-Qian Feng, Tao Yang, Kai Liu, Wen-Jing Cao, Ming-Ju Zhou, Chao Zhang, Ji-Yuan Zhang, Yan-Mei Jiao, Jin-Wen Song, Xing Fan, Ming Shi, Ruonan Xu and Fu-Sheng Wang declare that they have no conflict of interest.

Ethical approval

Ethical approvals were obtained from the Fifth Medical Center of Chinese PLA General Hospital, and the study was performed in accordance with the 1975 Declaration of Helsinki. Written informed consent was obtained from each participant prior to blood and liver sampling.

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Li, HJ., Yang, N., Mu, X. et al. Reduction of natural killer cells is associated with poor outcomes in patients with hepatitis B virus-related acute-on-chronic liver failure. Hepatol Int 16, 1398–1411 (2022). https://doi.org/10.1007/s12072-022-10386-9

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