Accumulation of T-bet–expressing T lymphocytes in the LP of patients with Crohn's disease. (A and B) Analysis of T-bet expression in the LP from patients with IBD and control patients. Colon cryosections were stained with a T-bet antibody and sections were analyzed by immunofluorescense microscopy and confocal laser microscopy, respectively. Many T-bet positive cells were seen in patients with Crohn's disease but not ulcerative colitis. Data are representative of four to five patients per group. (C) Immunohistochemical double staining analysis of the cellular expression of T-bet plus CD3 in consecutive cross sections. Cryosections from patients with Crohn's disease (n = 10), ulcerative colitis (n = 4) and control patients (n = 8) were stained with antibodies to CD3 and T-bet, as indicated. One representative experiment is shown. Double positive cells are indicated by arrows. (D) Quantitative analysis of T-bet-positive CD3+ LP T cells in patients with Crohn's disease, ulcerative colitis, and control patients. The percentage of double-positive T cells in four to six patients per group was quantified as specified in Materials and Methods. Data represents mean values ± SD. (E) Analysis of T-bet expression by LPMCs from patients with Crohn's disease and control patients. Nuclei were counterstained with DAPI, as specified in Materials and Methods. Most T cells from control patients showed weak expression of T-bet in perinuclear areas, whereas high cytoplasmic and nuclear expression was noted in patients with Crohn's disease. (F) Double staining analysis for GATA-3 and CD3 in the LP of patients with Crohn's disease (CD) and control patients (con), as indicated. Colon cryosection were stained with a polyclonal anti–GATA-3 antibody and anti-CD3 antibodies. There was a marked reduction of GATA-3 expression in LP T cells from patients with Crohn's disease compared with control patients. Data are representative of four to six patients per group. Cytospins from PMA-plus ionomycin-stimulated Jurkat cells served as a positive control.
Accumulation of T-bet–expressing T lymphocytes in the LP of patients with Crohn's disease. (A and B) Analysis of T-bet expression in the LP from patients with IBD and control patients. Colon cryosections were stained with a T-bet antibody and sections were analyzed by immunofluorescense microscopy and confocal laser microscopy, respectively. Many T-bet positive cells were seen in patients with Crohn's disease but not ulcerative colitis. Data are representative of four to five patients per group. (C) Immunohistochemical double staining analysis of the cellular expression of T-bet plus CD3 in consecutive cross sections. Cryosections from patients with Crohn's disease (n = 10), ulcerative colitis (n = 4) and control patients (n = 8) were stained with antibodies to CD3 and T-bet, as indicated. One representative experiment is shown. Double positive cells are indicated by arrows. (D) Quantitative analysis of T-bet-positive CD3+ LP T cells in patients with Crohn's disease, ulcerative colitis, and control patients. The percentage of double-positive T cells in four to six patients per group was quantified as specified in Materials and Methods. Data represents mean values ± SD. (E) Analysis of T-bet expression by LPMCs from patients with Crohn's disease and control patients. Nuclei were counterstained with DAPI, as specified in Materials and Methods. Most T cells from control patients showed weak expression of T-bet in perinuclear areas, whereas high cytoplasmic and nuclear expression was noted in patients with Crohn's disease. (F) Double staining analysis for GATA-3 and CD3 in the LP of patients with Crohn's disease (CD) and control patients (con), as indicated. Colon cryosection were stained with a polyclonal anti–GATA-3 antibody and anti-CD3 antibodies. There was a marked reduction of GATA-3 expression in LP T cells from patients with Crohn's disease compared with control patients. Data are representative of four to six patients per group. Cytospins from PMA-plus ionomycin-stimulated Jurkat cells served as a positive control.
