High-dimensional analysis of IL-10–dependent regulation of the antiviral response in the lung and the colon. High-dimensional analysis of IL-10–dependent.

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High-dimensional analysis of IL-10–dependent regulation of the antiviral response in the lung and the colon. High-dimensional analysis of IL-10–dependent regulation of the antiviral response in the lung and the colon. Mass cytometric analysis of cells recovered from lungs (A, C, and E) or colons (B, D, and F) of γHV68-infected B6 or IL-10KO mice harvested at 9 dpi. Files were normalized and gated on viable, single CD45+ cells (defined as 191Ir+193Ir+195Pt−89CD45+) prior to analysis, where numbers indicate isotopic mass for each measured parameter. (A and B) PhenoGraph analysis of CD45+ cells from (A) lungs or (B) colons of γHV68-infected B6 or IL-10KO mice. Clustering was done on 4344 events per file (34,752 total), with clustering in the lung based on 34 markers (29 clusters identified; Table IV) and clustering in the colon based on 36 markers (20 clusters identified; Table V); lungs and colons were harvested from separate cohorts. PhenoGraph-defined clusters are colored according to cluster identifier. (C and D) Definition of cellular phenotypes across PhenoGraph-defined clusters according to the indicated lineage markers, with distinct cell types given unique colors. CD45+ MHC II+ and CD45+ MHC II− clusters were defined by exclusion from other phenotypes. (E and F) Identification of clusters with statistically significant differences in frequency between B6 and IL-10KO infected (E) lungs and (F) colons, showing cluster frequencies (left panel) and expression for the identified parameters within the identified cluster (right panel). In the right panel of (E), plotted events are CD4+ T cells identified in (C). In the right panel of (F), plotted events are the dominant CD64+ cell clusters identified in (D). Parameters with a different maximum scale value between B6 and IL-10KO are identified by italicized text and an asterisk. Data are from γHV68-infected B6 and IL-10KO lungs (n = 4 mice per genotype) and colons (n = 4 mice per genotype) harvested 9 dpi. Data depict mean ± SEM, with individual symbols indicating values from independent samples. All samples were analyzed for statistical significance using unpaired t tests, corrected for multiple comparisons using the Holm-Sidak method. **p < 0.01, ***p < 0.001. Abigail K. Kimball et al. ImmunoHorizons 2019;3:94-109 Copyright © 2019 The Authors