[en] In horses, the mechanisms of lipopolysaccharide (LPS) stimulation of isolated neutrophils to produce reactive oxygen species remain unknown. We re-investigated this problem by monitoring the luminol-enhanced chemiluminescence (CL) produced by LPS-stimulated equine neutrophils. The neutrophils were isolated from horse blood by discontinuous density gradient centrifugation (> or = 99% neutrophils; viability > or = 98%). Increasing concentrations of Escherichia coli (E. coli) LPS (from 0.01-10 microg ml(-1)) were used to activate the neutrophils. When LPS was used directly, without another stimulator, the respiratory burst of neutrophils was not activated (N=12 horses; n=5 assays per horse). On the contrary, when LPS was added to whole blood, the neutrophils isolated from this blood were stimulated in a LPS dose-dependent manner, but polymyxin B added to whole blood suppressed this stimulation (N=2; n=6). LPS dissolved in autologous equine plasma stimulated the isolated neutrophils in a dose-dependent manner from 0.1-10 microg ml(-1) (N=5; n=12). Heat inactivation of the plasma abolished this CL increase (N=2; n=5). LPS added to equine albumin did not stimulate the isolated neutrophils (N=2; n=5). On the contrary, the addition of gamma-globulins (1 mg ml(-1)) to LPS (10 microg ml(-1)) led to the stimulation of neutrophils (N=2; n=5). We concluded that LPS did not directly stimulate the isolated equine neutrophils, but that plasmatic factors are needed for the stimulation of these cells by LPS.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Benbarek, H.
Deby-Dupont, G.
Caudron, I.
Grulke, Sigrid ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Département clinique des animaux de compagnie et des équidés
Deby, Christiane ; Centre Hospitalier Universitaire de Liège - CHU > Administration des patients - Admission des hospitalisés
Lamy, Maurice ; Université de Liège - ULiège > Département des sciences cliniques > Anesthésie et réanimation
Serteyn, Didier ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Language :
English
Title :
Interactions between Lipopolysaccharides and Blood Factors on the Stimulation of Equine Polymorphonuclear Neutrophils
Aida Y., Pabst M.J. Priming of neutrophils by lipopolysaccharide for enhanced release of superoxide. Requirement for plasma but not for tumor necrosis factor-α Immunol. 145:1990;3017-3025.
Aida Y., Kusumoto K., Nakatomi K., Takada H., Pabst M.J., Maeda K. An analogue of lipid-A and LPS from Rhodobacter sphaeroides inhibits neutrophil responses to LPS by blocking receptor recognition of LPS and by depleting LPS-binding protein in plasma. J. Leukoc. Biol. 58:1995;675-682.
Benbarek H., Deby-Dupont G., Deby C., Caudron I., Mathy-Hartert M., Lamy M., Serteyn D. An experimental model based on chemiluminescence for the study of the activation of isolated equine leucocytes. Res. Vet. Sciences. 61:1996;59-64.
Benbarek H., Deby-Dupont G., Caudron I., Deby C., Lamy M., Serteyn D. Failure of lipopolysaccharides to directly trigger the chemiluminescence response of the isolated equine polymorphonuclear leucocytes. Vet. Res. Comm. 21:1997;477-482.
Berczi I., Bertok L., Bereznai T. Comparative studies on the toxicity of Escherichia coli- lipopolysaccharide endotoxin in various animal species. Can. J. Microbiol. 12:1966;1070-1071.
Bochsler, P.N., Slauson, D.O., Neilsen, N.R., 1992. Secretory activity of equine polymorphonuclear leukocytes: Stimulus specificity and priming effects of bacterial lipopolysaccharide, Vet. Immunol. Immunopathol. 31, pp. 241-253.
Briheim G., Stendahl O., Dahlgren C. Intra-and extracellular events in luminol-dependent chemiluminescence of polymorphonuclear leucocytes. Infect. Immun. 45:1984;1-5.
Burrows G.E. Dose-response of ponies to parenteral Escherichia coli endotoxin. Can. J. Comp. Med. 45:1981;207-210.
David S.A., Balaram P., Mathan V.I. Characterization of the interaction of lipid-A and lipopolysaccharide with human serum albumin: Implications for an endotoxin carrier function for albumin. J. Endotox. Res. 2:1995;99-106.
Dorn C.R., Coffman J.R., Schmidt D.A., Garner H.E., Addison J.B., McCune E.L. Neutropenia and salmonellosis in hospitalized horses. J. Am. Vet. Med. Assoc. 166:1975;65-67.
Forehand J.R., Pabst M.J., Phillips W.A., Johnston R.B. Jr. Lipopolysaccharide priming of human neutrophils for an enhanced respiratory burst. Role of intracellular free calcium. J. Clin. Invest. 83:1989;74-83.
Fujishima S., Aikawa N. Neutrophil-mediated tissue injury and its modulation. Int. Care Med. 21:1995;277-285.
Henricks P.A.J., van der Tol M.E., Thyssen R.M., van Asbeck B.S., Verhoef J. Escherichia coli-lipopolysaccharides diminish and enhance cell function of polymorphonuclear leukocytes. Infect. Immun. 15:1983;294-301.
Hunt R.J., Allen D., Moore J.N. Effect of the endotoxin on equine digital hemodynamics and starling forces. Am. J. Vet. Res. 19:1987;25-28.
Meyers K., Reed S., Keck M., Clem M., Baily W. Circulating endotoxin-like substance(s) and altered hemostasis in horse with gastrointestinal disorders: An interim report. Am. J. Vet. Res. 43:1982;2233-2238.
Moore R.A., Bates N.C., Hancock R.E. Interaction of polycationic antibiotics with Pseudomonas aeruginosa-lipopolysaccharide and lipid-A studied by using dansyl-polymyxin. Antimicrob. Agents Chemother. 29:1986;496-500.
Nicotra J., Orsini A.J., Debari V.A. Chemiluminescence response of the human polymorphonuclear neutrophil to lipopolysaccharides. Cell. Biophys. 7:1985;283-291.
Proctor R.A. Endotoxin in vitro interactions with human neutrophils: Depression of chemiluminescence, oxygen consumption, superoxide production, and killing. Infect. Immun. 25:1979;912-921.
Pycock J.F., Allen E., Morris T.H. Rapid, single-step isolation of equine neutrophils on a discontinuous Percoll density gradient. Res. Vet. Sciences. 42:1987;411-412.
Sample A.K., Czuprynski C.J. Priming and stimulation of bovine neutrophils by recombinant human interleukin-1 alpha and tumor necrosis factor alpha. J. Leukoc. Biol. 49:1991;107-115.
Sample A.K., Czuprynski C.J. Bovine neutrophils chemiluminescence is preferentially stimulated by homologous IL-1, but inhibited by the human IL-1 receptor antagonist. Vet. Immunol. Immunopathol. 41:1994;165-172.
Seethanathan P., Bottoms G.D., Schafer K. Characterization of release of tumor necrosis factor, interleukin-1, and superoxide anion from equine white blood cells in response to endotoxin. Am. J. Vet. Res. 51:1990;1221-1225.
Shapira L., Champagne C., Gordon B., Amar S., Van Dyke T.E. Lipopolysaccharide priming of superoxide release by human neutrophils: Role of membrane CD14 and serum LPS binding protein. Inflammation. 19:1995;289-295.
Tool A.T.J., Pabst M.J., Roos D., Verhoeven A.J. Phosphatidyl-inositol-linked CD14 is involved in priming of human neutrophils by lipopolysaccharide (LPS), but not in the inactivation of LPS. Prog. Clin. Biol. Res. 388:1994;137-145.
Vosbeck K., Tobias P., Mueller H., Allen R.A., Arfors K.E., Ulevitch R.J., Sklar L.A. Priming of polymorphonuclear granulocytes by lipopolysaccharides and its complexes with lipopolysaccharide binding protein and high density lipoprotein. J. Leukoc. Biol. 47:1990;97-104.
Ward D.S., Fessler J.F., Bottoms G.D., Turek J. Equine endotoxemia: Cardiovascular, eicosanoid, hematologic, blood chemical, and plasma enzyme alterations. Am. J. Vet. Res. 48:1987;1150-1156.
Webster, R.O., Hong, S.R., Johnstan, R.B. Jr., Henson, P.M., 1980. Biological effects of the human complement fragments C5a and C5b desarg on neutrophil function. Immunopharmacology 2, 201-219.
Wilson M.E., Jones D.P., Munkenbeck P., Morrison D.C. Serum-dependent and independent effects of bacterial lipopolysaccharides on human neutrophil oxidative capacity in vitro. J. Reticuloendothel. Soc. 31:1982;43-57.
Wright S.D., Ramos R.A., Patel M., Miller D.S. Septin: A factor in plasma that opsonizes lipopolysaccharide-bearing particles for recognition by CD14 on phagocytes. J. Exp. Med. 176:1992;719-727.
Zheng H., Crowley J.J., Chan J.C., Raffin T.A. Attenuation of LPS-induced neutrophil thomboxane B2 release and chemiluminescence. J. Cell. Physiol. 146:1991;164-269.