Volume 133, Issue 6, Pages 1882-1892 (December 2007) Aggravation of Different Types of Experimental Colitis by Depletion or Adhesion Blockade of Neutrophils  Anja A. Kühl, Hacer Kakirman, Markus Janotta, Stefan Dreher, Philipp Cremer, Nina N. Pawlowski, Christoph Loddenkemper, Markus M. Heimesaat, Katja Grollich, Martin Zeitz, Stefan Farkas, Jörg C. Hoffmann  Gastroenterology  Volume 133, Issue 6, Pages 1882-1892 (December 2007) DOI: 10.1053/j.gastro.2007.08.073 Copyright © 2007 AGA Institute Terms and Conditions

Figure 1 Representative picture for hemorrhagic secretion around the eyes and nose of rats with TNBS-induced colitis because of anti-L-selectin mAb treatment (OX85). Gastroenterology 2007 133, 1882-1892DOI: (10.1053/j.gastro.2007.08.073) Copyright © 2007 AGA Institute Terms and Conditions

Figure 2 Increased mortality of rats with TNBS-induced colitis because of mAb treatment with OX85 (dashed line, n = 20) and HRL3 (dotted line, n = 10) compared with TS2/9-treated controls (solid line, n = 20). Shown is the Kaplan–Meier analysis. Gastroenterology 2007 133, 1882-1892DOI: (10.1053/j.gastro.2007.08.073) Copyright © 2007 AGA Institute Terms and Conditions

Figure 3 Severe colitis with necrosis in rats with TNBS-induced colitis because of anti-L-selectin mAb treatment. Representative picture of patchy necrosis (arrow) upon OX85 treatment. Gastroenterology 2007 133, 1882-1892DOI: (10.1053/j.gastro.2007.08.073) Copyright © 2007 AGA Institute Terms and Conditions

Figure 4 Increased mortality of rats with TNBS-induced colitis because of mAb treatment with HRL4 (dashed line, n = 10) compared with TS2/9-treated controls (solid line, n = 10). Shown is the Kaplan–Meier analysis. Gastroenterology 2007 133, 1882-1892DOI: (10.1053/j.gastro.2007.08.073) Copyright © 2007 AGA Institute Terms and Conditions

Figure 5 In vivo microscopy of colon mucosa in rats with TNBS-induced colitis upon anti-L-selectin mAb treatment. In animals receiving (A) control mAb TS2/9, massive leukocyte invasion can be observed, (B) nonblocking mAb HRL4 leukocytes still invade into colon mucosa whereas (C) blocking mAb OX85 and (D) blocking HRL3 leukocytes invasion almost completely. Gastroenterology 2007 133, 1882-1892DOI: (10.1053/j.gastro.2007.08.073) Copyright © 2007 AGA Institute Terms and Conditions

Figure 6 Increased mortality of rats with DNBS-induced colitis because of neutrophil depletion with anti-rat PMN (dashed line, n = 8) compared with anti-rat IgG-treated controls (solid line, n = 8) and shams (dotted line, n = 4). Shown is the Kaplan–Meier analysis. Gastroenterology 2007 133, 1882-1892DOI: (10.1053/j.gastro.2007.08.073) Copyright © 2007 AGA Institute Terms and Conditions

Figure 7 Increased rat DNBS-induced colitis because of neutrophil depletion. Representative picture of longitudinally opened colons of control antibody-treated (A) and neutrophil-depleted rats (B). Control colon is of normal appearance without evidence of inflammation (macroscopic score, 0), whereas colon of neutrophil-depleted rat exhibits shortening, stenosis with prestenotic dilatation, and ulceration (macroscopic score, 4). (C) Box blot analysis of the macroscopic score of all PMN-depleted rats (anti-rat PMN, n = 8) compared with controls (n = 8). (D) Histologic examination revealed transmural infiltration, cryptic abscesses (arrow), and loss of epithelium in colon of neutrophil-depleted rats (histologic score, 4) resulting (E) in significantly increased histologic scores in the box blot analysis. (F) More severe inflammation leads to significant colon shortening. Gastroenterology 2007 133, 1882-1892DOI: (10.1053/j.gastro.2007.08.073) Copyright © 2007 AGA Institute Terms and Conditions

Figure 8 Increased mortality of mice with established CD4+CD45RBhigh-induced transfer colitis because of neutrophil depletion. After first signs of colitis at day 14, IP application of antineutrophil mAb RB6-8C5 (dashed line, n = 5) or mouse IgG (solid line, n = 5) was started. Arrows indicate the IP application of RB6-8C5 and control IgG, respectively. Shown is the Kaplan–Meier analysis. Gastroenterology 2007 133, 1882-1892DOI: (10.1053/j.gastro.2007.08.073) Copyright © 2007 AGA Institute Terms and Conditions

Figure 9 Increased bacterial load in rats with DNBS-induced colits because of neutrophil depletion. (A) Viable aerobic bacteria are increased in luminal colon contents of neutrophil-depleted rats (ND, open circles, n = 4) compared with control IgG-treated rats (C, diamonds, n = 6). Colony-forming units (CFU) per gram luminal content of E coli (solid symbols), Proteus species (shaded symbols), and gram-positive cocci (mainly Enterococci; open symbols) are shown, with squares indicating mean values. (B) Some viable anaerobic bacterial species are increased in luminal colon contents of neutrophil-depleted rats compared with control rats: Clostridium species (shaded symbols) and total bacterial load (open symbols) but not Bacteroides/Prevotella species (solid symbols). Gastroenterology 2007 133, 1882-1892DOI: (10.1053/j.gastro.2007.08.073) Copyright © 2007 AGA Institute Terms and Conditions