Mucosal FOXP3+ regulatory T cells are numerically deficient in acute and chronic GvHD by Kathrin Rieger, Christoph Loddenkemper, Jochem Maul, Thomas Fietz,

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Mucosal FOXP3+ regulatory T cells are numerically deficient in acute and chronic GvHD by Kathrin Rieger, Christoph Loddenkemper, Jochem Maul, Thomas Fietz, Daniel Wolff, Harald Terpe, Beate Steiner, Erika Berg, Stephan Miehlke, Martin Bornhäuser, Thomas Schneider, Martin Zeitz, Harald Stein, Eckhard Thiel, Rainer Duchmann, and Lutz Uharek Blood Volume 107(4):1717-1723 February 15, 2006 ©2006 by American Society of Hematology

Comparison of the number of CD8+ T cells per 10 hpf's in intestinal biopsies. Comparison of the number of CD8+ T cells per 10 hpf's in intestinal biopsies. Paraffin-embedded colon biopsies of healthy controls (n = 13), and patients after allogeneic transplantation with GvHD (n = 62; chronic GvHD, n = 18; acute GvHD, n = 44), CMV-colitis (n = 6), and diverticulitis (n = 6) were stained for CD8 by immunohistochemistry. The number of CD8+ cells was counted per 10 high-power fields. Results are expressed as mean ± SEM. The differences of CD8+ T cells in GvHD, CMV, and diverticulitis were not statistically significant. Kathrin Rieger et al. Blood 2006;107:1717-1723 ©2006 by American Society of Hematology

Histology and immunohistochemistry for CD4+, CD8+, and CD3+FOXP3+ T cells of representative colonic biopsies from healthy controls and patients with no GvHD, with GvHD after bone marrow transplantation, and with CMV infection. Histology and immunohistochemistry for CD4+, CD8+, and CD3+FOXP3+ T cells of representative colonic biopsies from healthy controls and patients with no GvHD, with GvHD after bone marrow transplantation, and with CMV infection. A normal amount of CD3+ T cells (red, membranous) with very few regulatory T cells coexpressing FOXP3 (brown, nuclear) is found in healthy controls, whereas mucosal biopsies of patients without GvHD display a high number of CD3+FOXP3+ T cells in the lamina propria (arrows, bottom row) as opposed to a low number of regulatory T cells in patients with GvHD. All FOXP3+ cells in addition expressed CD25 (inset/bottom row). The number of CD8+ T cells is increased in biopsies from patients with GvHD especially in and around crypts displaying many characteristic apoptotic bodies. In CMV infection (inset/upper panel: nuclear staining for CMV antigen), the number of CD3+FOXP3+ regulatory T cells is also elevated compared with patients with GvHD. Original magnifications, × 200; and × 600 (inset). Microscope: Olympus AX70 (Olympus, Melville, NY); numeric aperture of objective lenses: × 20, 0.70 mm; × 60, 1.40. Stains: hematoxylin and eosin (H&E; top row); APAAP and immunoperoxide. Camera: JVC KY-F70 (JVC, Yokohama, Japan); acquisition software: DISKUS (Koenigswinter, Germany); and software used for image processing: Adobe Photoshop 7.0 (Adobe Systems, San Jose, CA). Kathrin Rieger et al. Blood 2006;107:1717-1723 ©2006 by American Society of Hematology

Number of FOXP3+ Tregs per 100 CD8+ lymphocytes in healthy controls, and patients without GvHD, with acute and chronic GvHD, with CMV-colitis, and with diverticulitis. Number of FOXP3+ Tregs per 100 CD8+ lymphocytes in healthy controls, and patients without GvHD, with acute and chronic GvHD, with CMV-colitis, and with diverticulitis. Paraffin-embedded intestinal biopsies of healthy controls (n = 13), and patients after allogeneic transplantation without GvHD (n = 33), with acute GvHD (n = 44), with chronic GvHD (n = 18), with CMV-colitis (n = 6), and with diverticulitis (n = 6) were immunohistochemically stained for CD8 and costained with anti-CD3/anti-FOXP3. The number of CD8+ cells and CD3+/FOXP3+ cells was counted per 10 high-power fields. Results are expressed as mean ± SEM. The ratio of FOXP3+ Tregs per 100 CD8+ lymphocytes was significantly increased in samples without GvHD versus acute and chronic GvHD. Kathrin Rieger et al. Blood 2006;107:1717-1723 ©2006 by American Society of Hematology