Anti-inflammatory effects of C1-Inhibitor in porcine and human whole blood are independent of its protease inhibition activity

Abstract

C1-Inhibitor (C1-INH) is an important biological inhibitor, regulating several protein cascade systems. Recent research has shown that the molecule exhibits properties not dependent on its protease inhibition activity. Serum and whole blood from pigs and humans were pre-incubated with C1-INH, iC1-INH or the complement inhibitors SPICE or compstatin. Whole, live Escherichia coli were then added for further incubation. Complement activation, a range of cytokines, chemokines and growth factors, as well as the leukocyte activation markers wCD11R3 (pig) and CD11b (human) were measured. Both C1-INH and iC1-INH dose-dependently and significantly (P<0.05) reduced a range of E. coli-induced pro-inflammatory cytokines and chemokines in porcine and human whole blood, as well as growth factors in human whole blood. Differences between the two forms of C1-INH and between the two species were modest. Most of these anti-inflammatory effects could not be explained by complement inhibition, as specific complement inhibitors had minor effect on several of the mediators. C1-Inhibitor had no inhibitory effect on E. coli-induced complement activation, while iC1-INH enhanced complement activation. The presented data indicate that C1-INH has broad anti-inflammatory effects in E. coli-induced inflammation in pig and human whole blood. These effects are largely independent of the protease inhibition activity.

Keywords

C1-Inhibitor complement cytokines E. coli iC1-Inhibitor inflammation

References

Caliezi C. , Wuillemin WA , Zeerleder S. , Redondo M. , Eisele B. , Hack CE C1-esterase inhibitor: an anti-inflammatory agent and its potential use in the treatment of diseases other than hereditary angioedema. Pharmacol Rev 2000; 52: 91-112.

Kirschfink M. Targeting complement in therapy. Immunol Rev 2001; 180: 177-189.

Horstick G. , Heimann A. , Gotze O. et al. Intracoronary application of C1 esterase inhibitor improves cardiac function and reduces myocardial necrosis in an experimental model of ischemia and reperfusion. Circulation 1997; 95: 701-708.

Nielsen EW , Mollnes TE , Harlan JM , Winn RK C1-inhibitor reduces the ischaemia-reperfusion injury of skeletal muscles in mice after aortic cross-clamping. Scand J Immunol 2002; 56: 588-592.

De Simoni MG , Rossi E. , Storini C. , Pizzimenti S. , Echart C. , Bergamaschini L. The powerful neuroprotective action of C1-inhibitor on brain ischemia-reperfusion injury does not require C1q. Am J Pathol 2004; 164: 1857-1863.

Longhi L. , Perego C. , Ortolano F. et al. C1-inhibitor attenuates neurobehavioral deficits and reduces contusion volume after controlled cortical impact brain injury in mice. Crit Care Med 2009; 37: 659-665.

Fiane AE , Videm V. , Johansen HT , Mellbye OJ , Nielsen EW , Mollnes TE C1-inhibitor attenuates hyperacute rejection and inhibits complement, leukocyte and platelet activation in an ex vivo pig-to-human perfusion model . Immunopharmacology 1999; 42: 231-243.

Kalter ES , van Dijk WC , Timmerman A. , Verhoef J. , Bouma BN Activation of purified human plasma prekallikrein triggered by cell wall fractions of Escherichia coli and Staphylococcus aureus. J Infect Dis 1983; 148: 682-691.

Ward PA The dark side of C5a in sepsis. Nat Rev Immunol 2004; 4: 133-142.

10.

Jansen PM , Eisele B. , de Jong I. et al. Effect of C1 inhibitor on inflammatory and physiologic response patterns in primates suffering from lethal septic shock. J Immunol 1998; 160: 475-484.

11.

Liu D. , Cai S. , Gu X. , Scafidi J. , Wu X. , Davis AE III. C1 inhibitor prevents endotoxin shock via a direct interaction with lipopolysaccharide . J Immunol 2003; 171: 2594-2601.

12.

Liu D. , Gu X. , Scafidi J. , Davis AE III. N-linked glycosylation is required for c1 inhibitor-mediated protection from endotoxin shock in mice. Infect Immun 2004 ; 72: 1946-1955.

13.

Liu D. , Cramer CC , Scafidi J. , Davis AE III. N-linked glycosylation at Asn3 and the positively charged residues within the amino-terminal domain of the c1 inhibitor are required for interaction of the C1 inhibitor with Salmonella enterica serovar Typhimurium lipopolysaccharide and lipid A. Infect Immun 2005; 73: 4478-4487.

14.

Cai S. , Davis AE III. Complement regulatory protein C1 inhibitor binds to selectins and interferes with endothelial-leukocyte adhesion. J Immunol 2003; 171: 4786-4791.

15.

Cai S. , Dole VS , Bergmeier W. et al. A direct role for C1 inhibitor in regulation of leukocyte adhesion . J Immunol 2005; 174: 6462-6466.

16.

Liu D. , Lu F. , Qin G. , Fernandes SM , Li J. , Davis AE III C1 inhibitor-mediated protection from sepsis. J Immunol 2007; 179: 3966-3972.

17.

Mollnes TE , Brekke OL , Fung M. et al. Essential role of the C5a receptor in E. coli-induced oxidative burst and phagocytosis revealed by a novel lepirudin-based human whole blood model of inflammation. Blood 2002; 100: 1869-1877.

18.

Rosengard AM , Liu Y. , Nie Z. , Jimenez R. Variola virus immune evasion design: expression of a highly efficient inhibitor of human complement. Proc Natl Acad Sci USA 2002; 99: 8808-8813.

19.

Sfyroera G. , Katragadda M. , Morikis D. , Isaacs SN , Lambris JD Electrostatic modeling predicts the activities of orthopoxvirus complement control proteins . J Immunol 2005; 174: 2143-2151.

20.

Katragadda M. , Magotti P. , Sfyroera G. , Lambris JD Hydrophobic effect and hydrogen bonds account for the improved activity of a complement inhibitor, compstatin. J Med Chem 2006; 49: 4616-4622.

21.

Mollnes TE , Lea T. , Froland SS , Harboe M. Quantification of the terminal complement complex in human plasma by an enzyme-linked immunosorbent assay based on monoclonal antibodies against a neoantigen of the complex. Scand J Immunol 1985; 22: 197-202.

22.

Mollnes TE , Redl H. , Hogasen K. et al. Complement activation in septic baboons detected by neoepitope-specific assays for C3b/ iC3b/C3c, C5a and the terminal C5b-9 complement complex (TCC) . Clin Exp Immunol 1993; 91: 295-300.

23.

Davis III AE , Mejia P. , Lu F. Biological activities of C1 inhibitor. Mol Immunol 2008; 45: 4057-4063.

24.

Nielsen EW , Waage C. , Fure H. et al. Effect of supra-physiologic levels of C1-inhibitor on the classical, lectin and alternative pathways of complement. Mol Immunol 2007; 44: 1829-1836.

25.

Kerr FK , Thomas AR , Wijeyewickrema LC et al. Elucidation of the substrate specificity of the MASP-2 protease of the lectin complement pathway and identification of the enzyme as a major physiological target of the serpin, C1-inhibitor. Mol Immunol 2008; 45: 670-677.

26.

Jiang H. , Wagner E. , Zhang H. , Frank MM Complement 1 inhibitor is a regulator of the alternative complement pathway . J Exp Med 2001; 194: 1609-1616.

27.

Kobayashi C. , Matsunami K. , Omori T. et al. Features of a newly cloned pig C1 esterase inhibitor. J Biochem (Tokyo) 2006; 140: 421-427.

28.

Thorgersen EB , Ghebremariam YT , Thurman JM et al. Candidate inhibitors of porcine complement . Mol Immunol 2007; 44: 1837-1844.

29.

Ziccardi RJ Spontaneous activation of the first component of human complement (C1) by an intramolecular autocatalytic mechanism. J Immunol 1982; 128: 2500-2504.

30.

Salvesen B. , Nielsen EW , Harboe M. , Saugstad OD , Mollnes TE Mechanisms of complement activation and effects of C1-inhibitor on the meconium-induced inflammatory reaction in human cord blood. Mol Immunol 2009; 46: 688-694.

31.

Ziccardi RJ Demonstration of the interaction of native C1 with monomeric immunoglobulins and C1 inhibitor. J Immunol 1985; 134: 2559-2563.

32.

Thorgersen EB , Pharo A. , Haverson K. et al. Complement- and CD14-inhibition attenuate Escherichia coli-induced inflammatory response in porcine whole blood. Infect Immun 2009 ; 77: 725-732.

33.

Brekke OL , Christiansen D. , Fure H. et al. Combined inhibition of complement and CD14 abolish E. coli-induced cytokine-, chemokine- and growth factor-synthesis in human whole blood. Mol Immunol 2008; 45: 3804-3813.

34.

Yano K. , Liaw PC , Mullington JM et al. Vascular endothelial growth factor is an important determinant of sepsis morbidity and mortality. J Exp Med 2006; 203: 1447-1458.

35.

Hamilton JA , Anderson GP GM-CSF biology. Growth Factors 2004; 22: 225-231.

36.

von Vietinghoff S. , Ley K. Homeostatic regulation of blood neutrophil counts. J Immunol 2008; 181: 5183-5188.

37.

Spight D. , Trapnell B. , Zhao B. , Berclaz P. , Shanley TP Granulocyte-macrophage-colony-stimulating factor-dependent peritoneal macrophage responses determine survival in experimentally induced peritonitis and sepsis in mice. Shock 2008; 30: 434-442.

38.

Basu S. , Dunn AR , Marino MW et al. Increased tolerance to endotoxin by granulocyte-macrophage colony-stimulating factor-deficient mice. J Immunol 1997; 159: 1412-1417.

39.

Zhang H. , Li J. , Barrington RA , Liang G. , Qin G. , Liu DX An anti-endotoxin peptide that generates from the amino-terminal domain of complement regulatory protein C1 inhibitor. Biochem Biophys Res Commun 2007; 359: 285-291.