Abstract
Toll-like receptors (TLRs) have been previously shown to mediate oxidative burst in chicken heterophils. This study was conducted to begin to map the molecular pathways that regulate TLR-mediated oxidative burst. Peripheral blood heterophils from neonatal chicks were isolated and exposed to known inhibitors of signal transduction pathways for either 20 min (genistein, verapamil, or chelerythrine) or 120 min (pertussis toxin) at 39°C. The cells were then stimulated for 30 min at 39°C with Salmonella enteritidis lipopolysaccharide (LPS) or Staphylococcus aureus lipoteichoic acid (LTA). The heterophil oxidative burst was then quantitated by luminol-dependent chemiluminescence (LDCL). Genistein (a tyrosinekinase inhibitor), verapamil (a calcium channel blocker), chelerythrine (a protein kinase C inhibitor), and pertussis toxin (a G-protein inhibitor) significantly reduced LPS-stimulated oxidative burst in chicken heterophils by 34, 50, 63, and 51%, respectively. Although genistein had a statistically significant effect on reducing LPS-stimulated LDCL biologically it seems to play only a minor role within the oxidative burst pathway. Heterophils stimulated with the gram-positive TLR agonist, LTA, activated a different signal transduction pathway since chelerythrine was the only inhibitor that significantly reduced (72%) LTA-stimulated oxidative burst. These findings demonstrate that distinct signal transduction pathways differentially regulate the stimulation of oxidative burst in avian heterophils. Pertussis toxin-sensitive, protein kinase C-dependent, Ca++-dependent G proteins appear to regulate oxidative burst of avian heterophils stimulated with gram-negative agonist LPS; whereas, a protein kinase C-dependent signal transduction pathway plays the major role activating the oxidative burst of avian heterophils stimulated with gram-positive agonists. The distinct differences in the response of heterophils to these two agonists illustrate the specificity of TLRs to pathogen-associated molecular patterns (PAMP)s.
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References
Harmon, B. G. 1998. Avian heterophils in inflammation and disease resistance. Poult. Sci. 77:972–977.
Kogut, M. H., G. I. Tellez, E. D. McGruder, B. M. Hargis, J. D. Williams, D. E. Corrier, and J. R. DeLoach. 1994. Heterophils are decisive components in the early responses of chickens to Salmonella enteritidis infections. Microb. Pathol. 16:141–151.
Kogut, M. H., K. J. Genovese, and V. K. Lowry. 2001. Differential activation of signal transduction pathways mediating phagocytosis, oxidative burst, and degranulation by chicken heterophils in response to stimulation with opsonized Salmonella enteritidis. Inflammation 25(1):7–15.
Kogut, M. H., E. D. McGruder, B. M. Hargis, D. E. Corrier, and J. R. DeLoach. 1995. In vivo activation of heterophil function in chickens following injection with Salmonella enteritidis—immune lymphokines. J. Leukoc. Biol. 57:56–62.
Stabler, J. G., T. McCormick, K. Powell, and M. H. Kogut. 1994. Avian heterophils and monocytes: Phagocytic and bactericidal activities against Salmonella enteritidis. Vet. Microbiol. 38:293–305.
Medzhitov, R. and C. A. Janeway Jr. 1997. Innate immunity: The virtues of a nonclonal system of recognition. Cell 91:295–298.
Schwandner, R., R. Dziarski, H. Wesche, M. Rothe, and C. J. Kirschning. 1999. Peptidoglycan-and lipoteichoic acid-induced cell activation is mediated by Toll-like receptor 2. J. Biol. Chem. 274(18):17406–17409.
Medzhitov, R. and C. A. Janeway Jr. 1997. Innate immunity: Impact on the adaptive immune response. Curr. Opin. Immunol. 9:4–9.
Janeway, C. A., Jr. and R. Medzhitov. 1999. Innate immunity: Lipoproteins take their Toll on the host. Curr. Biol. 9:R879-R882.
Medzhitov, R. and C. A. Janeway Jr. 2000. The Toll receptor family and microbial recognition. Trends Microbiol. 8(10):452–456.
Akira, S., K. Takeda., and T. Kaisho. 2001. Toll-like receptors: Critical proteins linking innate and acquired immunity. Nature Immunol. 2(8):675–680.
Boyd, Y., M. Goodchild, S. Morroll, and N. Bumstead. 2001. Mapping of the chicken and mouse genes for Toll-like receptor 2 (TLR2) to an evolutionarily conserved chromosomal segment. Immunogenetics 52:294–298.
Fukui, A., N. Inoue, M. Matsumoto, M. Nomura, K. Yamada, Y. Matsuda, K. Toyoshima, and T. Seya. 2001. Molecular cloning and functional characterization of chicken Toll-like receptors. J. Biol. Chem. 276(50):47143–47149.
Farnell, M. B., T. L. Crippen, H. He, C. L. Swaggerty, and M. H. Kogut. 2003. Oxidative burst mediated by Toll-like receptors (TLR) and CD14 on avian heterophils stimulated with bacterial Toll agonists. Dev. Comp. Immunol. 27(5):423–429.
Aliprantis, A. O., D. S. Weiss, and A. Zychlinsky. 2001. Toll-like receptor-2 transduces signals for NF-kappa B activation, apoptosis and reactive oxygen species production. J. Endotoxin Res. 7(4):287–291.
Aliprantis, A. O., R. B. Yang, M. R. Mark, S. Suggett, B. Devaux, J. D. Radolf, G. R. Klimpel, P. Godowski, and A. Zychlinsky. 1999. Cell activation and apoptosis by bacterial lipoproteins through Toll-like receptor-2. Science 285(5428):736–739.
National Research Council. 1994. Nutrient Requirements of Poultry, 9th edn. National Academy Press, Washington, DC.
Merrill, G. A., R. Bretthauer, J. Wright-Hicks, and R. C. Allen. 1996. Oxygenation activities of chicken polymorphonuclear leukocytes investigated by selective chemiluminigenic probes. Lab. Anim. Sci. 46(5):530–538.
Raloff, J. 1998. Livestock's role in antibiotic resistance. Sci. News 154(3):39.
Ferber, D. 2002. Livestock feed ban preserves drugs' power. Science 295(5552):27–28.
Lowenthal, J. W., B. Lambrecht, T. P. van den Berg, M. E. Andrew, A. D. G. Strom, and A. G. D. Bean. 2000. Avian cytokines—The natural approach to therapeutics. Dev. Comp. Immunol. 24:355–365.
Lodish, H., A. Berk, S. L. Zipursky, D. Matsudaira, D. Baltimore, and J. E. Darnell. 2000. Cell-to-cell signaling: Hormones and receptors. In: Molecular Cell Biology, 4th edn., Ch. 20. Freeman, New York, pp. 848–909.
Janeway, C. A., P. Travers, M. Walport, and M. Shlomchik. 2001. Signaling through immune system receptors. In: Immunobiology: The Immune System in Health and Disease, 5th edn., Ch. 6. Garland, New York, pp. 187–220.
Yu, P. W. and C. J. Czuprynski. 1996. Regulation of luminol-dependent chemiluminescence and degranulation by bovine neutrophils stimulated with opsonized zymosan. Vet. Immunol. Immunopathol. 50(1/2):29–42.
Ortiz-Carranza, O. and C. J. Czuprynski. 1992. Activation of bovine neutrophils by Pasteurella haemolytica leukotoxin is calcium dependent. J. Leukoc. Biol. 52(5):558–564.
Laudanna, C., D. Mochly-Rosen, T. Liron, G. Constantin, and E. C. Butcher. 1998. Evidence of zeta protein C involvement in polymorphonuclear neutrophil integrin-dependent adhesion and chemotaxis. J. Biol. Chem. 273(46):30306–30315.
Tilton, B., M. Andjelkovic, S. A. Didichenko, B. A. Hemmings, and M. Thelen. 1997. G-protein-coupled receptors and Fc gamma-receptors mediate activation of Akt/protein kinase B in human phagocytes. J. Biol. Chem. 272(44):28096–28101.
Crockett-Torabi, E. and J. C. Fantone. 1990. Soluble and insoluble immune complexes activate human neutrophil NADPH oxidase by distinct Fc gamma receptor-specific mechanisms. J. Immunol. 145(9):3026–3032.
Kogut, M., V. K. Lowry, and M. Farnell. 2002. Selective pharmacological inhibitors reveal the role of Syk tyrosine kinase, phospholipase C, phosphatidylinositol-3′-kinase, and p38 mitogen-activated protein kinase in Fc receptor-mediated signaling of chicken heterophil degranulation. Int. Immunopharmacol. 2(7):963–973.
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Farnell, M.B., He, H. & Kogut, M.H. Differential Activation of Signal Transduction Pathways Mediating Oxidative Burst by Chicken Heterophils in Response to Stimulation with Lipopolysaccharide and Lipoteichoic Acid. Inflammation 27, 225–231 (2003). https://doi.org/10.1023/A:1025088514676
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DOI: https://doi.org/10.1023/A:1025088514676