Figure 2.
A targeted candidate protein approach identifies a predicted phenotype of RA-responsive cells in the context of increased microenvironmental IL-23. (A) Mononuclear cell staining for T-cell lineage markers, transcription factors, and the cytokines IL-23 and IL-33 and their receptors using conventional IHC in the same GI biopsy cohort. Scale bars, 100 µm. Positive cells (brown) are stained by DAB, SG (CD4) or AEC (CD8) chromogen. Cells are counterstained with hematoxylin. Insets show higher manifaction with positice cells identified with arrows in gray (CD4) or red (all other markers). (B) CD8 and CD4 T-cell numbers were enumerated automatically using the Ariol system in subcrypt regions of upper and lower GI biopsies from allo-HSCT patients with and without histologic GVHD. (C) Mononuclear cell numbers were enumerated automatically using the Ariol system expressing T-cell transcription factors, cytokines, and cytokine receptors in subcrypt regions of upper and lower GI biopsies from allo-HSCT patients with and without histologic GVHD. (D) Volcano plot depicting fold change and statistical significance of RARαhi mononuclear cell numbers and number of cells expressing T-cell lineage markers, transcription factors, cytokines, and cytokine receptors in subcrypt regions of upper and lower GI biopsies from allo-HSCT patients with and without histologic GVHD. (E) Total cellular IL-23 (calculated as the product of the number of IL-23+ mononuclear cells and the mean intensity of staining of cells for IL-23) in subcrypt regions of upper and lower GI biopsies from allo-HSCT patients with and without histologic GVHD. (F) Volcano plot depicting correlation coefficients and statistical significance of numbers of RARαhi mononuclear cells and cells expressing T-cell lineage markers, transcription factors, cytokines, and cytokine receptors in subcrypt regions of upper and lower GI biopsies from allo-HSCT patients. P and r values are for Pearson correlations. Cells expressing CD8, T-bet, and IL-23R (circled) were significantly positively correlated with numbers of RARαhi mononuclear cells. Horizontal lines are medians. P > .10 indicates nonsignificant values. *P < .05; ** P < .01, Mann-Whitney U tests (A-E).

A targeted candidate protein approach identifies a predicted phenotype of RA-responsive cells in the context of increased microenvironmental IL-23. (A) Mononuclear cell staining for T-cell lineage markers, transcription factors, and the cytokines IL-23 and IL-33 and their receptors using conventional IHC in the same GI biopsy cohort. Scale bars, 100 µm. Positive cells (brown) are stained by DAB, SG (CD4) or AEC (CD8) chromogen. Cells are counterstained with hematoxylin. Insets show higher manifaction with positice cells identified with arrows in gray (CD4) or red (all other markers). (B) CD8 and CD4 T-cell numbers were enumerated automatically using the Ariol system in subcrypt regions of upper and lower GI biopsies from allo-HSCT patients with and without histologic GVHD. (C) Mononuclear cell numbers were enumerated automatically using the Ariol system expressing T-cell transcription factors, cytokines, and cytokine receptors in subcrypt regions of upper and lower GI biopsies from allo-HSCT patients with and without histologic GVHD. (D) Volcano plot depicting fold change and statistical significance of RARαhi mononuclear cell numbers and number of cells expressing T-cell lineage markers, transcription factors, cytokines, and cytokine receptors in subcrypt regions of upper and lower GI biopsies from allo-HSCT patients with and without histologic GVHD. (E) Total cellular IL-23 (calculated as the product of the number of IL-23+ mononuclear cells and the mean intensity of staining of cells for IL-23) in subcrypt regions of upper and lower GI biopsies from allo-HSCT patients with and without histologic GVHD. (F) Volcano plot depicting correlation coefficients and statistical significance of numbers of RARαhi mononuclear cells and cells expressing T-cell lineage markers, transcription factors, cytokines, and cytokine receptors in subcrypt regions of upper and lower GI biopsies from allo-HSCT patients. P and r values are for Pearson correlations. Cells expressing CD8, T-bet, and IL-23R (circled) were significantly positively correlated with numbers of RARαhi mononuclear cells. Horizontal lines are medians. P > .10 indicates nonsignificant values. *P < .05; ** P < .01, Mann-Whitney U tests (A-E).

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