Volume 116, Issue 4, Pages 874-883 (April 1999) VCAM-1 and ICAM-1 mediate leukocyte-endothelial cell adhesion in rat experimental colitis  Miquel Sans, Julián Panés, Esther Ardite, J.Ignasi Elizalde, Yolanda Arce, Montserrat Elena, Antonio Palacín, J.Carlos Fernández– Checa, Donald C. Anderson, Roy Lobb, Josep M. Piqué  Gastroenterology  Volume 116, Issue 4, Pages 874-883 (April 1999) DOI: 10.1016/S0016-5085(99)70070-3 Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 1 Expression of (A) ICAM-1 and (B) VCAM-1 relative to endothelial surface area in control rats and colitic animals at different time points after treatment with TNBS/ethanol. Endothelial surface area was estimated by binding of anti-ACE MAb 9B9, ICAM-1 expression by binding of anti–ICAM-1 MAb 1A29, and VCAM-1 expression by binding of anti–VCAM-1 MAb 5F10. Results are calculated as μg ICAM-1/μg 9B9 and μg VCAM-1/μg 9B9. *P < 0.01 vs. control. Gastroenterology 1999 116, 874-883DOI: (10.1016/S0016-5085(99)70070-3) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 2 Immunohistochemical preparation of a sample of colonic tissue from a colitic rat after intravenous administration of the anti–VCAM-1 MAb 5F10. Incubation with a secondary antibody, showing marked staining limited to the luminal side of the vessel, confirms that the anti–VCAM-1 MAb binds to the luminal endothelium (original magnification 400×). Gastroenterology 1999 116, 874-883DOI: (10.1016/S0016-5085(99)70070-3) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 3 Leukocyte (A) rolling and (B) adhesion in colonic venules. Control animals had a very low number of rolling or adherent leukocytes. A significant increment in leukocyte-endothelial cell interactions was observed at days 1 and 7 after induction of colitis and returned to basal values by 21 days. *P < 0.05 vs. control; #P < 0.001 vs. control and colitis 21 days. Gastroenterology 1999 116, 874-883DOI: (10.1016/S0016-5085(99)70070-3) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 4 Effects of immunoneutralization of ICAM-1 and VCAM-1 on leukocyte (A) rolling and (B) adhesion in animals studied 7 days after induction of colitis. Immunoneutralization of either ICAM-1 or VCAM-1 had no effects on leukocyte rolling. However, blocking of ICAM-1 significantly attenuated leukocyte adhesion, whereas blocking of VCAM-1 completely abrogated leukocyte adhesion in colonic venules of colitic rats. Isotype-matched control MAbs P-23 and UPC-10 had no effect. *P < 0.05 vs. control; #P < 0.05 vs. colitis plus saline. Gastroenterology 1999 116, 874-883DOI: (10.1016/S0016-5085(99)70070-3) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 5 Effect of treatment with anti–VCAM-1 MAb or the isotype-matched MAb UPC-10 on TNBS-induced colitis. Treatment with 5F10 significantly attenuated the decrease in body weight at days 1 and 2 after induction of colitis and induced a rapid recovery of body weight in the following days. ○, TNBS + 5F10; ■, TNBS + UPC-10. *P < 0.05, #P < 0.001; vs. UPC-10–treated colitic rats. Gastroenterology 1999 116, 874-883DOI: (10.1016/S0016-5085(99)70070-3) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 6 Macroscopic appearance of colon from 5F10 and UPC-10 treated rats. (A) Extensive ulceration involving most of the distal colon, with marked thickening of the bowel wall (macroscopic damage, 9), in a TNBS-colitic rat treated with UPC-10 (right). (B) Treatment of TNBS-induced colitis with 5F10 results in marked reduction of macroscopic damage, with discrete erythema and superficial ulceration, limited to 3 cm of distal colon (macroscopic damage score, 5). Gastroenterology 1999 116, 874-883DOI: (10.1016/S0016-5085(99)70070-3) Copyright © 1999 American Gastroenterological Association Terms and Conditions