Figure 2.

Lymphocytes with a regulatory phenotype are expanded in the colonic lamina propria [LP] during ulcerative colitis and lack αE integrin. Dual stain IHC was performed on formalin-fixed paraffin-embedded colonic biopsies to evaluate co-expression of the αE integrin [red] with FOXP3 [green]. Single and dual expressing cells [yellow, indicated by arrowheads] were enumerated using the Nuance Multispectral Tissue Imaging System combined with automated morphometric analysis. [A] A representative Nuance spectrally unmixed image is shown for a patient with active UC. Scale bar represents 50 μm. Automatic cell counting was used to enumerate [B] FOXP3+αE+, [C] FOXP3+αE− cells and [D] FOXP3−αE+ cells per mm² of cell area within the epithelium or LP in control subjects [n = 16] and patients with active UC [n = 14]. In a separate study cohort, FACS-sorted unstimulated CD4+αE+ and CD4+αE− colonic lymphocytes were analysed for TReg gene expression in [E] control subjects [n = 8] and [F] patients with active UC [n = 10].