Longitudinal liver sampling in patients with chronic hepatitis B starting antiviral therapy reveals hepatotoxic CD8+ T cells
Shirin Nkongolo, … , Harry L.A. Janssen, Adam J. Gehring
Shirin Nkongolo, … , Harry L.A. Janssen, Adam J. Gehring
Published January 3, 2023
Citation Information: J Clin Invest. 2023;133(1):e158903. https://doi.org/10.1172/JCI158903.
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Research Article Immunology Infectious disease

Longitudinal liver sampling in patients with chronic hepatitis B starting antiviral therapy reveals hepatotoxic CD8+ T cells

Abstract

Accumulation of activated immune cells results in nonspecific hepatocyte killing in chronic hepatitis B (CHB), leading to fibrosis and cirrhosis. This study aims to understand the underlying mechanisms in humans and to define whether these are driven by widespread activation or a subpopulation of immune cells. We enrolled CHB patients with active liver damage to receive antiviral therapy and performed longitudinal liver sampling using fine-needle aspiration to investigate mechanisms of CHB pathogenesis in the human liver. Single-cell sequencing of total liver cells revealed a distinct liver-resident, polyclonal CD8+ T cell population that was enriched at baseline and displayed a highly activated immune signature during liver damage. Cytokine combinations, identified by in silico prediction of ligand-receptor interaction, induced the activated phenotype in healthy liver CD8+ T cells, resulting in nonspecific Fas ligand–mediated killing of target cells. These results define a CD8+ T cell population in the human liver that can drive pathogenesis and a key pathway involved in their function in CHB patients.

Authors

Shirin Nkongolo, Deeqa Mahamed, Adrian Kuipery, Juan D. Sanchez Vasquez, Samuel C. Kim, Aman Mehrotra, Anjali Patel, Christine Hu, Ian McGilvray, Jordan J. Feld, Scott Fung, Diana Chen, Jeffrey J. Wallin, Anuj Gaggar, Harry L.A. Janssen, Adam J. Gehring

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Figure 5

The combination of IL-2 and IL-12 induces hepatotoxic CD8+ T cells in healthy donor intrahepatic cells.

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The combination of IL-2 and IL-12 induces hepatotoxic CD8+ T cells in he...
(A) IHMCs from 6 healthy donors were treated with IL-2, IL-12, or IL-2 plus IL-12 for 24 hours before quantification of IFN-γ and FasL expression of CXCR6+CD8+ T cells by flow cytometry. Circles and triangles indicate individual donors; bars indicate mean values. Statistical significance was assessed by 2-tailed, ratio-paired t test. (B) A multicolor flow cytometry panel was used to analyze IHMCs from 6 healthy donors, with and without 24 hours of treatment with IL-2 plus IL-12. Single, live CD3+ lymphocytes were selected before clustering. The hepatotoxic CD8+ T cell population, corresponding to baseline FNA samples from CHB patients, was only found in the treated healthy donor IHMCs. Heatmaps show median expression. (C) Source of IL2 and IL12 in CHB patients’ livers at baseline. A targeted scRNA-Seq assay was used to enrich for cytokine genes. Both IL2 and IL12A/IL12B could be detected.