Volume 122, Issue 4, Pages 1122-1132 (April 2002) Leukocyte recruitment in colon cancer: Role of cell adhesion molecules, nitric oxide, and transforming growth factor β1  Xavier Bessa, J.Ignasi Elizalde, Francesc Mitjans, Virgínia Piñol, Rosa Miquel, Julián Panés, Jaime Piulats, Josep M. Piqué, Antoni Castells  Gastroenterology  Volume 122, Issue 4, Pages 1122-1132 (April 2002) DOI: 10.1053/gast.2002.32369 Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 1 Schematic of the intravital microscopy system. In an anesthetized animal, a portion of the intestine is exteriorized through a laparotomy (for tumor examination, a skin flap is created and the tumor placed over a glass-covered window in the microscopy stage). Light produced by a mercury lamp is projected to the excitation filter (420–490 nm) and through the objective lens to the tissue surface. Reflected light is filtered by the emission filter (520 nm), and the image is digitized by the CCD camera. The video signal is recorded and later analyzed. Gastroenterology 2002 122, 1122-1132DOI: (10.1053/gast.2002.32369) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 2 Leukocyte-endothelium interactions in tumor, peritumor, and colonic venules at (A) baseline and (B) after LPS stimulation. □, rolling (leukocytes/30 seconds); ■, adhesion (leukocytes/100 μm). Results are expressed as mean ± standard deviation. #P < 0.05 vs. tumor microvessels. Gastroenterology 2002 122, 1122-1132DOI: (10.1053/gast.2002.32369) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 3 Effects of ICAM-1 and VCAM-1 immunoneutralization on (A) leukocyte adhesion and (B) rolling in mice studied 4 hours after administration of LPS. □, tumor vessels; ■, peritumor vessels. Results are expressed as mean ± standard deviation. #P < 0.05 vs. LPS alone. Gastroenterology 2002 122, 1122-1132DOI: (10.1053/gast.2002.32369) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 4 Correlation of endothelial ICAM-2 expression in tumor tissue with tumor weight. Accumulated activity was expressed as nanograms of anti–ICAM-2 binding per gram of tissue. Gastroenterology 2002 122, 1122-1132DOI: (10.1053/gast.2002.32369) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 5 Expression of (A) ICAM-1, (B) VCAM-1, (C) E-selectin, and (D) P-selectin corrected by the endothelial surface area in tumor vessels (□) and colonic vessels (■) at baseline and after administration of LPS. Results are expressed as mean ± standard deviation. #P < 0.05 vs. baseline value; $P < 0.05 vs. tumor vessels. Gastroenterology 2002 122, 1122-1132DOI: (10.1053/gast.2002.32369) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 6 Effect of (A) nonselective COX and (B) selective COX-2 inhibition on leukocyte rolling in tumor and peritumor venules. □, baseline values; ■, COX inhibition. Results are expressed as mean ± standard deviation. #P < 0.05 vs. tumor venules; $P < 0.05 vs. baseline. Gastroenterology 2002 122, 1122-1132DOI: (10.1053/gast.2002.32369) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 7 Effect of nonselective NO synthase inhibition on (A) leukocyte rolling and (B) adhesion after administration of LPS in animals pretreated with L-NAME (10 mg/kg intravenously). Results are expressed as mean ± standard deviation. Administration of L-NAME induced a dose-dependent increase in leukocyte rolling in both tumor (r = 0.45, P < 0.002 for trend) and peritumor (r = 0.35, P < 0.01 for trend) vasculature. #P < 0.05 vs. tumor vessels; $P < 0.05 vs. baseline and LPS challenge. Gastroenterology 2002 122, 1122-1132DOI: (10.1053/gast.2002.32369) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 8 Effect of TGF-β1 immunoblockade on (A) leukocyte rolling and (B) adhesion after administration of LPS in animals pretreated with an mAb against TGF-β1. Results are expressed as mean ± standard deviation. #P < 0.05 vs. tumor vessels; $P < 0.05 vs. baseline and LPS challenge. Gastroenterology 2002 122, 1122-1132DOI: (10.1053/gast.2002.32369) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 9 TGF-β1 immunohistochemical expression in experimental colon cancer. Gastroenterology 2002 122, 1122-1132DOI: (10.1053/gast.2002.32369) Copyright © 2002 American Gastroenterological Association Terms and Conditions