Volume 117, Issue 2, Pages 378-387 (August 1999) Surgically induced leukocytic infiltrates within the rat intestinal muscularis mediate postoperative ileus  Jörg C. Kalff, Timothy M. Carlos, Wolfgang H. Schraut, Timothy R. Billiar, Richard L. Simmons, Anthony J. Bauer  Gastroenterology  Volume 117, Issue 2, Pages 378-387 (August 1999) DOI: 10.1053/gast.1999.0029900378

1 Fig. 1. Time course of expression of ICAM-1 (■) and P-selectin (●) mRNA in the intestinal muscularis after surgical manipulation of the small intestine. Message levels were measured by RT-PCR, and signal was quantified using laser densitometry. Upper panels show typical acrylamide gel RT-PCR product bands for ICAM-1 and P-selectin. Lower plot shows percent increase from control of quantified data for both adhesion molecule mRNAs. ICAM-1 shows early induction at 3 hours and a second sustained period of expression at 12 hours postoperatively. P-selectin shows a similar pattern, but with a lesser degree of induction. Data are expressed as percent of controls in arbitrary units (n = 4). Gastroenterology 1999 117, 378-387DOI: (10.1053/gast.1999.0029900378)

2 Fig. 2. Fluorescence micrographs of muscularis whole mounts from a surgically manipulated rat stained with rat-specific monoclonal antibodies against ICAM-1, P-selectin, and LFA-1. (A) Early signal of ICAM-1 in endothelial cells of the muscularis microvasculature at 3 hours. (B) ICAM-1 signal was also found on infiltrating cells 12 hours after manipulation. LFA-1 was expressed on infiltrating and resident cells beginning 1 hour postoperatively, shown at 3 hours in C. (C) Strong LFA-1 signal found on the round to oblong–shaped monocytes and PMNs. A weaker but noticeable signal was also present on the stellate resident macrophages. (D) A strong P-selectin immunofluorescent signal was seen on endothelial cells at 3 hours. (A and C, original magnification 200×; B and D, original magnification 250×). Gastroenterology 1999 117, 378-387DOI: (10.1053/gast.1999.0029900378)

3 Fig. 3. Myeloperoxidase staining for PMNs within muscularis whole mounts. (A) Low basal presence of PMNs in control animals. (B) Surgical manipulation resulted in massive recruitment of PMNs into the muscularis 24 hours postoperatively. (C) This infiltration observed at 24 hours was significantly reduced by adhesion molecule antibody treatment (1A29, WT.1, and WT.3) of the surgically manipulated animals (original magnification 100×). Gastroenterology 1999 117, 378-387DOI: (10.1053/gast.1999.0029900378)

4 Fig. 4. Histogram of infiltrating leukocytes in muscularis whole mounts from control rats and surgically manipulated rats with and without adhesion-positive molecule antibody (mAb) treatment. Myeloperoxidase-positive neutrophils (2), monocytes (ED1 [▨]), and mast cells (■, avidin-positive) were histochemically and immunohistochemically stained and counted at 200× magnification. Data are expressed as means ± SEM (n = 5-7). Gastroenterology 1999 117, 378-387DOI: (10.1053/gast.1999.0029900378)

5 Fig. 5. Bethanechol-stimulated dose-response curves of circular muscle contractile activity generated from jejunal circular muscle of control rats (●), manipulated rats (▴), and manipulated rats that received antibody treatment (■). Data are expressed as means ± SEM (n = 5-7). Gastroenterology 1999 117, 378-387DOI: (10.1053/gast.1999.0029900378)

6 Fig. 6. Organ bath recorded mechanical traces of spontaneous circular muscle contractile activity. (A) Typical mechanical trace from a control animal. (B) Decrease in spontaneous contractile activity seen after manipulation. (C) Inhibition of leukocyte recruitment by monoclonal adhesion molecule antibodies prevented a decrease in spontaneous circular smooth muscle activity. Gastroenterology 1999 117, 378-387DOI: (10.1053/gast.1999.0029900378)