Eur Surg Res 2004;36:331–337

Attenuation of Leukocyte Sequestration by Selective Blockade of PECAM-1 or VCAM-1 in Murine Endotoxemia

Nolte D.a,b · Kuebler W.b,c · Muller W.d · Wolff K.a · Messmer K.b
aDepartment of Oral and Maxillofacial Surgery – Regional Plastic Surgery, Knappschaftskrankenhaus Bochum-Langendreer, University of Bochum, Bochum, bInstitute for Surgical Research, Ludwig Maximilian University of Munich, Munich, and cInstitute of Physiology, Charité – Universitätsmedizin Berlin, Berlin, Germany; dDepartment of Pathology and Laboratory Medicine, Weill Medical College, New York, N.Y., USA
email Corresponding Author

 goto top of outline Key Words

  • Adhesion molecules
  • Platelet-endothelial cell adhesion molecule 1
  • Vascular cell adhesion molecule 1
  • Salmonella abortus equi endotoxin
  • Leukocyte-endothelial cell interaction
  • Microcirculation

 goto top of outline Abstract

Background: Molecular mechanisms regulating leukocyte sequestration into the tissue during endotoxemia and/or sepsis are still poorly understood. This in vivo study investigates the biological role of murine PECAM-1 and VCAM-1 for leukocyte sequestration into the lung, liver and striated skin muscle. Methods: Male BALB/c mice were injected intravenously with murine PECAM-1 IgG chimera or monoclonal antibody (mAb) to VCAM-1 (3 mg/kg body weight); controls received equivalent doses of IgG2a (n = 6 per group). Fifteen minutes thereafter, 2 mg/kg body weight of Salmonella abortus equi endotoxin was injected intravenously. At 24 h after the endotoxin challenge, lungs, livers and striated muscle of skin were analyzed for their myeloperoxidase activity. To monitor intravital leukocyte-endothelial cell interactions, fluorescence videomicroscopy was performed in the skin fold chamber model of the BALB/c mouse at 3, 8 and 24 h after injection of endotoxin. Results: Myeloperoxidase activity at 24 h after the endotoxin challenge in lungs (12,171 ± 2,357 mU/g tissue), livers (2,204 ± 238 mU/g) and striated muscle of the skin (1,161 ± 110 mU/g) was significantly reduced in both treatment groups as compared to controls, with strongest attenuation in the PECAM-1 IgG treatment group. Arteriolar leukocyte sticking at 3 h after endotoxin (230 ± 46 cells × mm–2) was significantly reduced in both treatment groups. Leukocyte sticking in postcapillary venules at 8 h after endotoxin (343 ± 69 cells/mm2) was found reduced only in the VCAM-1-mAb-treated animals (215 ± 53 cells/mm2), while it was enhanced in animals treated with PECAM-1 IgG (572 ± 126 cells/mm2). Conclusion: These data show that both PECAM-1 and VCAM-1 are involved in endotoxin-induced leukocyte sequestration in the lung, liver and muscle, presumably through interference with arteriolar and/or venular leukocyte sticking.

Copyright © 2004 S. Karger AG, Basel

 goto top of outline References
  1. Dinarello CA: The proinflammatory cytokines interleukin-1 and tumor necrosis factor and treatment of the septic shock syndrome. J Infect Dis 1991;163:1177–1184.
  2. Vestweber D: Cytokine inducible CAMs on endothelial cells of the mouse. Eur Surg Res 1992;24:321–327.
  3. ten Cate H, Schoenmakers SH, Franco R, Timmerman JJ, Groot AP, Spek CA, Reitsma PH: Microvascular coagulopathy and disseminated intravascular coagulation. Crit Care Med 2001;29:S95–S97.

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  4. Muchamuel T, Menon S, Pisacane P, Howard MC, Cockayne DA: IL-13 protects mice from lipopolysaccharide-induced lethal endotoxemia: Correlation with down-modulation of TNF-alpha, IFN-gamma, and IL-12 production. J Immunol 1997;158:2898–2903.
  5. Li P, Allen H, Banerjee S, Franklin S, Herzog L, Johnston C, Mcdowell J, Paskind M, Rodman L, Salfeld J, Towne E, Tracey D, Wardwell S, Wei FY, Wong W, Kamen R, Seshadri T: Mice deficient in IL-1 beta-converting enzyme are defective in production of mature IL-1 beta and resistant to endotoxic shock. Cell 1995;80:401–411.
  6. Jin HK, Yang RH, Marsters SA, Bunting SA, Wurm FM, Chamow SM, Ashkenazi A: Protection against rat endotoxic shock by p55 tumor necrosis factor (TNF) receptor immunoadhesin: Comparison with anti-TNF monoclonal antibody. J Infect Dis 1994;170:1323–1326.
  7. Fisher CJ Jr, Agosti JM, Opal SM, Lowry SF, Balk RA, Sadoff JC, Abraham E, Schein RM, Benjamin E: Treatment of septic shock with the tumor necrosis factor receptor:Fc fusion protein. The soluble TNF receptor sepsis study group. N Engl J Med 1996;334:1697–1702.
  8. Osborn L, Hession C, Tizard R, Vassallo C, Luhowskyj S, Chi-Rosso G, Lobb R: Direct expression cloning of vascular cell adhesion molecule 1, a cytokine-induced endothelial protein that binds lymphocytes. Cell 1989;59:1203–1211.
  9. Vaporciyan AA, Delisser HM, Yan HC, Mendiguren II, Thom SR, Jones ML, Ward PA, Albelda SM: Involvement of platelet endothelial cell adhesion molecule-1 in neutrophil recruitment in vivo. Science 1993;262:1580–1582.
  10. Muller WA, Weigl SA, Deng X, Phillips DM: PECAM-1 is required for transendothelial migration of leukocytes. J Exp Med 1993;178:449–460.
  11. Albelda SM, Muller WA, Buck CA, Newman PJ: Molecular and cellular properties of PECAM-1 (endoCAM/CD31): A novel vascular cell-cell adhesion molecule. J Cell Biol 1991;114:1059–1068.
  12. Liao F, Huynh HK, Eiroa A, Greene T, Polizzi E, Muller WA: Migration of monocytes across endothelium and passage through extracellular matrix involve separate molecular domains of PECAM-1. J Exp Med 1995;182:1337–1343.
  13. Davies MJ, Gordon JL, Gearing AJH, Pigott R, Woolf N, Katz D, Kyriakopoulos A: The expression of the adhesion molecules ICAM-1, VCAM-1, PECAM, and E-selectin in human atherosclerosis. J Pathol 1993;171:223–229.
  14. Ilan N, Madri JA: PECAM-1: Old friend, new partners. Curr Opin Cell Biol 2003;15:515–524.
  15. Orosz CG, Ohye RG, Pelletier RP, Vanbuskirk AM, Huang E, Morgan C, Kincade PW, Ferguson RM: Treatment with antivascular cell adhesion molecule-1 monoclonal antibody induces long-term murine cardiac allograft acceptance. Transplantation 1993;56:453–460.
  16. Khan BV, Parthasarathy SS, Alexander RW, Medford RM: Modified low density lipoprotein and its constituents augment cytokine-activated vascular cell adhesion molecule 1 gene expression in human vascular endothelial cells. J Clin Invest 1995;95:1262–1270.
  17. Liao F, Ali J, Greene T, Muller WA: Soluble domain 1 of platelet-endothelial cell adhesion molecule (PECAM) is sufficient to block transendothelial migration in vitro and in vivo. J Exp Med 1997;185:1349–1357.
  18. Hahne M, Lenter M, Jäger U, Isenmann S, Vestweber D: VCAM-1 is not involved in LPAM-1 (alpha 4 beta p/alpha 4 beta 7) mediated binding of lymphoma cells to high endothelial venules of mucosa-associated lymph nodes. Eur J Cell Biol 1993;61:290–298.
  19. Kuebler WM, Abels C, Schuerer L, Goetz AE: Measurement of neutrophil content in brain and lung tissue by a modified myeloperoxidase assay. Int J Microcirc Clin Exp 1996;16:89–97.
  20. Nolte D, Hecht R, Schmid P, Botzlar A, Menger MD, Neumueller C, Sinowatz F, Vestweber D, Messmer K: Role of Mac-1 and ICAM-1 in ischemia-reperfusion injury in a microcirculation model of BALB/c mice. Am J Physiol 1994;36:H1320–H1328.
  21. Zeintl H, Sack F-U, Intaglietta M, Messmer K: Computer assisted leukocyte velocity measurement in intravital microscopy. Int J Microcirc Clin Exp 1989;8:293–302.
  22. Nolte D, Schmid P, Jäger U, Botzlar A, Roesken F, Hecht R, Uhl E, Messmer K, Vestweber D: Leukocyte rolling in venules of striated muscle and skin is mediated by P-selectin, not by L-selectin – Rapid communication. Am J Physiol 1994;36:H1637–H1642.
  23. Morise Z, Ueda M, Kitajima M, Epstein CJ, Granger DN, Grisham MB: Reactive oxygen species and vascular cell adhesion molecule-1 in distant organ failure following bile duct obstruction in mice. Dig Dis Sci 2002;47:607–613.
  24. Fleming SD, Anderson J, Wilson F, Shea- Donohue T, Tsokos GC: C5 is required for CD49d expression on neutrophils and VCAM expression on vascular endothelial cells following mesenteric ischemia/reperfusion. Clin Immunol 2003;106:55–64.
  25. Neumann B, Machleidt T, Lifka A, Pfeffer K, Vestweber D, Mak TW, Holzmann B, Kronke M: Crucial role of 55-kilodalton TNF receptor in TNF-induced adhesion molecule expression and leukocyte organ infiltration. J Immunol 1996;156:1587–1593.
  26. Lundberg AH, Granger N, Russell J, Callicutt S, Gaber LW, Kotb M, Sabek O, Gaber AO: Temporal correlation of tumor necrosis factor-alpha release, upregulation of pulmonary ICAM-1 and VCAM-1, neutrophil sequestration, and lung injury in diet-induced pancreatitis. J Gastrointest Surg 2000;4:248–257.
  27. Callicutt CS, Sabek O, Fukatsu K, Lundberg AH, Gaber L, Wilcox H, Kotb M, Gaber AO: Diminished lung injury with vascular adhesion molecule-1 blockade in choline-deficient ethionine diet-induced pancreatitis. Surgery 2003;133:186–196.
  28. Piedboeuf B, Gamache M, Frenette J, Horowitz S, Baldwin HS, Petrov P: Increased endothelial cell expression of platelet-endothelial cell adhesion molecule-1 during hyperoxic lung injury. Am J Respir Cell Mol Biol 1998;19:543–553.
  29. Christofidou-Solomidou M, Scherpereel A, Wiewrodt R, Ng K, Sweitzer T, Arguiri E, Shuvaev V, Solomides CC, Albelda SM, Muzykantov VR: PECAM-directed delivery of catalase to endothelium protects against pulmonary vascular oxidative stress. Am J Physiol Lung Cell Mol Physiol 2003;285:L283–L292.
  30. van Oosten M, van de Bilt E, de Vries HE, van Berkel TJ, Kuiper J: Vascular adhesion molecule-1 and intercellular adhesion molecule-1 expression on rat liver cells after lipopolysaccharide administration in vivo. Hepatology 1995;22:1538–1546.
  31. Essani NA, Bajt ML, Farhood A, Vonderfecht SL, Jaeschke H: Transcriptional activation of vascular cell adhesion molecule-1 gene in vivo and its role in the pathophysiology of neutrophil-induced liver injury in murine endotoxin shock. J Immunol 1997;158:5941–5948.
  32. Chosay JG, Essani NA, Dunn CJ, Jaeschke H: Neutrophil margination and extravasation in sinusoids and venules of liver during endotoxin-induced injury. Am J Physiol 1997;272:G1195–G1200.
  33. Chosay JG, Fisher MA, Farhood A, Ready KA, Dunn CJ, Jaeschke H: Role of PECAM-1 (CD31) in neutrophil transmigration in murine models of liver and peritoneal inflammation. Am J Physiol 1998;274:G776–G782.
  34. Neubauer K, Wilfling T, Ritzel A, Ramadori G: Platelet-endothelial cell adhesion molecule-1 gene expression in liver sinusoidal endothelial cells during liver injury and repair. J Hepatol 2000;32:921–932.
  35. Nolte D: Role of adhesion molecules in the development of septic organ dysfunction. Curr Pharm Design, in press.
  36. Hoffmann JN, Vollmar B, Inthorn D, Schildberg FW, Menger MD: The thrombin antagonist hirudin fails to inhibit endotoxin-induced leukocyte/endothelial cell interaction and microvascular perfusion failure. Shock 2000;14:528–534.
  37. Kunkel EJ, Jung U, Ley K: TNF-alpha induces selectin-mediated leukocyte rolling in mouse cremaster muscle arterioles. Am J Physiol 1997;272:H1391–H1400.
  38. Nolte D, Dehning B, Flemisch S, Muller WA, Galanos C, Messmer K: Blockade of platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31) prevents endotoxin-induced microvascular leak syndrome in striated muscle of the BALB/c mouse. Langenbecks Arch Chir Suppl Kongressbd 1997;114:639–642.

 goto top of outline Author Contacts

Dirk Nolte, MD, DDS, PhD
Department of Oral and Maxillofacial Surgery – Regional Plastic Surgery
University Clinic at the Ruhr University of Bochum, In der Schornau 23–25
DE–44892 Bochum (Germany)
Tel. +49 234 299 3502/12, Fax +49 234 299 3509, E-Mail

 goto top of outline Article Information

Received: May 26, 2004
Accepted after revision: June 25, 2004
Number of Print Pages : 7
Number of Figures : 3, Number of Tables : 0, Number of References : 38

 goto top of outline Publication Details

European Surgical Research (Clinical and Experimental Surgery)

Vol. 36, No. 6, Year 2004 (Cover Date: November-December 2004)

Journal Editor: K. Messmer, Munich
ISSN: 0014–312X (print), 1421–9921 (Online)

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