Highly multiplexed 2-dimensional imaging mass cytometry analysis of HBV-infected liver
Daniel Traum, … , Klaus H. Kaestner, Kyong-Mi Chang
Daniel Traum, … , Klaus H. Kaestner, Kyong-Mi Chang
Published February 23, 2021
Citation Information: JCI Insight. 2021;6(7):e146883. https://doi.org/10.1172/jci.insight.146883.
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Research Article Hepatology Infectious disease

Highly multiplexed 2-dimensional imaging mass cytometry analysis of HBV-infected liver

Abstract

Studies of human hepatitis B virus (HBV) immune pathogenesis are hampered by limited access to liver tissues and technologies for detailed analyses. Here, utilizing imaging mass cytometry (IMC) to simultaneously detect 30 immune, viral, and structural markers in liver biopsies from patients with hepatitis B e antigen+ (HBeAg+) chronic hepatitis B, we provide potentially novel comprehensive visualization, quantitation, and phenotypic characterizations of hepatic adaptive and innate immune subsets that correlated with hepatocellular injury, histological fibrosis, and age. We further show marked correlations between adaptive and innate immune cell frequencies and phenotype, highlighting complex immune interactions within the hepatic microenvironment with relevance to HBV pathogenesis.

Authors

Daniel Traum, Yue J. Wang, Kathleen B. Schwarz, Jonathan Schug, David K.H. Wong, Harry L.A. Janssen, Norah A. Terrault, Mandana Khalili, Abdus S. Wahed, Karen F. Murray, Phillip Rosenthal, Simon C. Ling, Norberto Rodriguez-Baez, Richard K. Sterling, Daryl T.Y. Lau, Timothy M. Block, Michael D. Feldman, Elizabeth E. Furth, William M. Lee, David E. Kleiner, Anna S. Lok, Klaus H. Kaestner, Kyong-Mi Chang

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Figure 5

PhenoGraph analysis identifies distinct immune subclusters that correlate with serum ALT.

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PhenoGraph analysis identifies distinct immune subclusters that correlat...
(A) PhenoGraph analysis of IMC data shown as: (left) a tSNE plot with 13 subclusters (S1–S13); (middle) a heatmap displaying expression of cellular markers; (right) 9 representative tSNE plots showing various marker expression. (B) Representative IMC images with PhenoGraph subcluster events shown as distinct dots (top); select antibody staining patterns in IMC images (middle); overlays of subcluster dots in red onto individual markers in green, with yellow color representing colocalization (bottom). Three images on the far left and bottom far right show background of HepPar1 in blue and collagen I in light gray to indicate hepatic architecture. (C) Scatter plots comparing cellular densities (cells/mm2 ROI) for immune subclusters on the x axis and manually gated CD45+ immune subsets on the y axis for 28 IA, 6 IT, and 10 NC subjects. Spearman’s correlation coefficients (rs) and P values are shown, with red font for P < 0.05. (D) Dot plots comparing hepatic densities of CD45-enriched subclusters between 28 IA and 6 IT subjects. Median values shown as red horizontal bars. P values by Mann-Whitney U tests with values < 0.00625 highlighted in red font. (E) Upper right half of the table shows Spearman’s correlation coefficients between immune subcluster densities as a heatmap (positive correlations in orange and negative correlations in blue) with corresponding P values in left bottom half, with significant P values highlighted in pink with red font (P < 0.00238 considered significant). (F) Scatter plots comparing subcluster densities (cells/mm2 ROI) on the x axis and serum ALT levels, HBV DNA, and Ishak histological scores on the y axis. Spearman’s correlation coefficients (rs) and P values are shown. Correlations with P < 0.0071 were considered significant and highlighted in red.