Figure 6.

Significant upregulation of gut-tropic α₄+β₇+CD4+IL23R+ T cells on IL-23 application. (A) CD4+ lamina propria mononuclear cells from patients with Crohn’s disease (CD) (n=10) were isolated with magnetic beads and left untreated or stimulated for 72 hours with TGF-β, IL-6 or IL-23 or a combination of all three cytokines. Expression of the IL23R on CD4+ cells was determined by FACS analysis. (B) CD4+ peripheral blood mononuclear cells from healthy controls (n=6) or patients with CD (n=13) were isolated with magnetic beads and left untreated or stimulated for 72 hours with TGF-β, IL23, a combination of both cytokines, IL12, IL17, IL21 or TNF. Expression of the IL23R on CD4+ cells among α₄−β₇− or α₄+β₇+ cells was determined by FACS analysis. (C) Whole intestinal biopsies from patients with CD were cultivated for 24 hours in an organ culture chamber at 37°C with 95% O₂/5% CO₂ atmosphere. Biopsies were untreated (n=10) or stimulated with 20 ng/mL IL-23 (n=10). Expression of the IL23R on CD4+ cells among α₄−β₇− or α₄+β₇+ cells was determined via flow cytometry. (D) Whole intestinal biopsies from patients with CD were cultivated for 24 hours in an organ culture chamber at 37°C with 95% O₂/5% CO₂ atmosphere. Biopsies were untreated, stimulated with 25 µg/mL anti-TNF antibody or 25 µg/mL anti-TNF antibody plus 20 ng/mL IL-23. Expression of CD4, IL23R and TNFR2 was determined via flow cytometry. Data represent mean values±SEM. *p≤0.05; **p≤0.01; ***p≤0.001.