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Vol. 101, No. 2, 2012
Issue release date: February 2012
Section title: Original Paper
Free Access
Neonatology 2012;101:140–146
(DOI:10.1159/000330567)

Toll-Like Receptor 1/2 Stimulation Induces Elevated Interleukin-8 Secretion in Polymorphonuclear Leukocytes Isolated from Preterm and Term Newborn Infants

Thornton N.L. · Cody M.J. · Yost C.C.
Department of Pediatrics and Program in Human Molecular Biology and Genetics, University of Utah, Salt Lake City, Utah, USA
email Corresponding Author

Abstract

Background: Neonatal neutrophil dysfunction contributes to inflammatory tissue damage in newborn infants. Toll-like receptors (TLRs) activate the innate immune response through recognition of pathogen-associated molecular patterns. Expression and function of TLRs by neonatal neutrophils has not well been characterized. Objective: We hypothesized that, compared to polymorphonuclear leukocytes (PMNs) isolated from adults, neonatal PMNs isolated from either term or preterm infants express and release different levels of inflammatory cytokines and chemokines in response to stimulation with TLR1–9 agonists. Methods: We stimulated PMNs isolated from preterm (n = 12) and term (n = 10) infants as well as adults (n = 10) with agonists recognized by TLRs1–9 and quantified chemokine and cytokine expression and secretion by ELISA and Luminex® multiplex quantification assay. Results: Neonatal and adult PMNs stimulated with agonists recognized by TLRs1–9 differentially secrete inflammatory products. Signaling via TLR2 heterodimers is a potent mechanism for release of interleukin-8, a critical proinflammatory chemokine, by neonatal PMNs – a previously unrecognized facet of neonatal inflammation. Following TLR1/2 (PAM3CSK4) stimulation, interleukin-8 secretion by neonatal PMNs, whether term or preterm, substantially exceeds that of adult PMNs assayed in parallel. Conclusions: These studies provide new insights relevant to the inflammatory biology of neonates, both term and preterm, and implicate exaggerated PMN recruitment in neonatal syndromes of dysregulated inflammation such as necrotizing enterocolitis or neonatal chronic lung disease.

© 2011 S. Karger AG, Basel


  

Key Words

  • Neutrophil
  • Innate immunity
  • Toll-like receptors
  • Interleukin-8
  • Inflammation

References

  1. Nathan C: Neutrophils and immunity: challenges and opportunities. Nat Rev Immunol 2006;6:173–182.
  2. Segal AW: How neutrophils kill microbes. Annu Rev Immunol 2005;23:197–223.
  3. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, Weinrauch Y, Zychlinsky A: Neutrophil extracellular traps kill bacteria. Science 2004;303:1532–1535.
  4. Yost CC, Cody MJ, Harris ES, Thornton NL, McInturff AM, Martinez ML, Chandler NB, Rodesch CK, Albertine KH, Petti CA, Weyrich AS, Zimmerman GA: Impaired neutrophil extracellular trap (net) formation: a novel innate immune deficiency of human neonates. Blood 2009;113:6419–6427.
  5. Kaufmann I, Hoelzl A, Schliephake F, Hummel T, Chouker A, Peter K, Thiel M: Polymorphonuclear leukocyte dysfunction syndrome in patients with increasing sepsis severity. Shock 2006;26:254–261.
  6. Aldridge AJ: Role of the neutrophil in septic shock and the adult respiratory distress syndrome. Eur J Surg 2002;168:204–214.
  7. Wipke BT, Allen PM: Essential role of neutrophils in the initiation and progression of a murine model of rheumatoid arthritis. J Immunol 2001;167:1601–1608.
  8. Fiocchi C: Inflammatory bowel disease: etiology and pathogenesis. Gastroenterology 1998;115:182–205.
  9. Coxon A, Cullere X, Knight S, Sethi S, Wakelin MW, Stavrakis G, Luscinskas FW, Mayadas TN: Fc gamma RIII mediates neutrophil recruitment to immune complexes. A mechanism for neutrophil accumulation in immune-mediated inflammation. Immunity 2001;14:693–704.
  10. Nguyen CN, Schnulle PM, Chegini N, Luo X, Koenig JM: Neonatal neutrophils with prolonged survival secrete mediators associated with chronic inflammation. Neonatology 2010;98:341–347.
  11. Koenig JM, Yoder MC: Neonatal neutrophils: the good, the bad, and the ugly. Clin Perinatol 2004;31:39–51.
  12. Carr R: Neutrophil production and function in newborn infants. Br J Haematol 2000;110:18–28.
  13. Angus DC, Linde-Zwirble WT, Lidicker J, Clermont G, Carcillo J, Pinsky MR: Epidemiology of severe sepsis in the united states: analysis of incidence, outcome, and associated costs of care. Crit Care Med 2001;29:1303–1310.
  14. Stoll BJ, Hansen N, Fanaroff AA, Wright LL, Carlo WA, Ehrenkranz RA, Lemons JA, Donovan EF, Stark AR, Tyson JE, Oh W, Bauer CR, Korones SB, Shankaran S, Laptook AR, Stevenson DK, Papile LA, Poole WK: Changes in pathogens causing early-onset sepsis in very-low-birth-weight infants. N Engl J Med 2002;347:240–247.
  15. Stoll BJ, Hansen NI, Higgins RD, Fanaroff AA, Duara S, Goldberg R, Laptook A, Walsh M, Oh W, Hale E: Very low birth weight preterm infants with early onset neonatal sepsis: the predominance of gram-negative infections continues in the national institute of child health and human development neonatal research network, 2002–2003. Pediatr Infect Dis J 2005;24:635–639.
  16. Hunter CJ, Upperman JS, Ford HR, Camerini V: Understanding the susceptibility of the premature infant to necrotizing enterocolitis (NEC). Pediatr Res 2008;63:117–123.
  17. Speer CP: Pulmonary inflammation and bronchopulmonary dysplasia. J Perinatol 2006;26(suppl 1):S57–S62; discussion S63–S54.
  18. Zentay Z, Sharaf M, Qadir M, Drafta D, Davidson D: Mechanism for dexamethasone inhibition of neutrophil migration upon exposure to lipopolysaccharide in vitro: role of neutrophil interleukin-8 release. Pediatr Res 1999;46:406–410.
  19. Takeuchi O, Akira S: Pattern recognition receptors and inflammation. Cell 2010;140:805–820.
  20. Akira S, Uematsu S, Takeuchi O: Pathogen recognition and innate immunity. Cell 2006;124:783–801.
  21. Takeda K, Akira S: Toll-like receptors in innate immunity. Int Immunol 2005;17:1–14.
  22. Wynn JL, Scumpia PO, Winfield RD, Delano MJ, Kelly-Scumpia K, Barker T, Ungaro R, Levy O, Moldawer LL: Defective innate immunity predisposes murine neonates to poor sepsis outcome but is reversed by TLR agonists. Blood 2008;112:1750–1758.
  23. Lindemann SW, Yost CC, Denis MM, McIntyre TM, Weyrich AS, Zimmerman GA: Neutrophils alter the inflammatory milieu by signal-dependent translation of constitutive messenger RNAs. Proc Natl Acad Sci U S A 2004;101:7076–7081.
  24. Yost CC, Denis MM, Lindemann S, Rubner FJ, Marathe GK, Buerke M, McIntyre TM, Weyrich AS, Zimmerman GA: Activated polymorphonuclear leukocytes rapidly synthesize retinoic acid receptor-alpha: a mechanism for translational control of transcriptional events. J Exp Med 2004;200:671–680.
  25. Caron JE, La Pine TR, Augustine NH, Martins TB, Hill HR: Multiplex analysis of toll-like receptor-stimulated neonatal cytokine response. Neonatology 2010;97:266–273.
  26. Scapini P, Lapinet-Vera JA, Gasperini S, Calzetti F, Bazzoni F, Cassatella MA: The neutrophil as a cellular source of chemokines. Immunol Rev 2000;177:195–203.
  27. Levy O, Suter EE, Miller RL, Wessels MR: Unique efficacy of toll-like receptor 8 agonists in activating human neonatal antigen-presenting cells. Blood 2006;108:1284–1290.
  28. Contrino J, Krause PJ, Slover N, Kreutzer D: Elevated interleukin-1 expression in human neonatal neutrophils. Pediatr Res 1993;34:249–252.
  29. Usmani SS, Kamran S, Harper RG, Wapnir RA, Mehta R: Effect of maternal labor and mode of delivery on polymorphonuclear leukocyte function in healthy neonates. Pediatr Res 1993;33:466–468.
  30. Molloy EJ, O’Neill AJ, Grantham JJ, Sheridan-Pereira M, Fitzpatrick JM, Webb DW, Watson RW: Labor promotes neonatal neutrophil survival and lipopolysaccharide responsiveness. Pediatr Res 2004;56:99–103.
  31. Weinberger B, Vetrano AM, Syed K, Murthy S, Hanna N, Laskin JD, Laskin DL: Influence of labor on neonatal neutrophil apoptosis, and inflammatory activity. Pediatr Res 2007;61:572–577.
  32. Molloy EJ, O’Neill AJ, Doyle BT, Grantham JJ, Taylor CT, Sheridan-Pereira M, Fitzpatrick JM, Webb DW, Watson RW: Effects of heat shock and hypoxia on neonatal neutrophil lipopolysaccharide responses: altered apoptosis, toll-like receptor-4 and CD11B expression compared with adults. Biol Neonate 2006;90:34–39.
  33. Liu Y, Zhu L, Fatheree NY, Liu X, Pacheco SE, Tatevian N, Rhoads JM: Changes in intestinal toll-like receptors and cytokines precede histological injury in a rat model of necrotizing enterocolitis. Am J Physiol Gastrointest Liver Physiol 2009;297:G442–G450.

  

Author Contacts

Christian Con Yost, MD
Department of Pediatrics/Neonatology, University of Utah
Williams Building, 295 Chipeta Way
Salt Lake City, UT 84108 (USA)
Tel. +1 801 581 7052, E-Mail christian.yost@u2m2.utah.edu

  

Article Information

Received: April 19, 2011
Accepted after revision: July 1, 2011
Published online: September 23, 2011
Number of Print Pages : 7
Number of Figures : 2, Number of Tables : 4, Number of References : 33

  

Publication Details

Neonatology (Fetal and Neonatal Research)

Vol. 101, No. 2, Year 2012 (Cover Date: February 2012)

Journal Editor: Halliday H.L. (Belfast), Speer C.P. (Würzburg)
ISSN: 1661-7800 (Print), eISSN: 1661-7819 (Online)

For additional information: http://www.karger.com/NEO


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References

  1. Nathan C: Neutrophils and immunity: challenges and opportunities. Nat Rev Immunol 2006;6:173–182.
  2. Segal AW: How neutrophils kill microbes. Annu Rev Immunol 2005;23:197–223.
  3. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, Weinrauch Y, Zychlinsky A: Neutrophil extracellular traps kill bacteria. Science 2004;303:1532–1535.
  4. Yost CC, Cody MJ, Harris ES, Thornton NL, McInturff AM, Martinez ML, Chandler NB, Rodesch CK, Albertine KH, Petti CA, Weyrich AS, Zimmerman GA: Impaired neutrophil extracellular trap (net) formation: a novel innate immune deficiency of human neonates. Blood 2009;113:6419–6427.
  5. Kaufmann I, Hoelzl A, Schliephake F, Hummel T, Chouker A, Peter K, Thiel M: Polymorphonuclear leukocyte dysfunction syndrome in patients with increasing sepsis severity. Shock 2006;26:254–261.
  6. Aldridge AJ: Role of the neutrophil in septic shock and the adult respiratory distress syndrome. Eur J Surg 2002;168:204–214.
  7. Wipke BT, Allen PM: Essential role of neutrophils in the initiation and progression of a murine model of rheumatoid arthritis. J Immunol 2001;167:1601–1608.
  8. Fiocchi C: Inflammatory bowel disease: etiology and pathogenesis. Gastroenterology 1998;115:182–205.
  9. Coxon A, Cullere X, Knight S, Sethi S, Wakelin MW, Stavrakis G, Luscinskas FW, Mayadas TN: Fc gamma RIII mediates neutrophil recruitment to immune complexes. A mechanism for neutrophil accumulation in immune-mediated inflammation. Immunity 2001;14:693–704.
  10. Nguyen CN, Schnulle PM, Chegini N, Luo X, Koenig JM: Neonatal neutrophils with prolonged survival secrete mediators associated with chronic inflammation. Neonatology 2010;98:341–347.
  11. Koenig JM, Yoder MC: Neonatal neutrophils: the good, the bad, and the ugly. Clin Perinatol 2004;31:39–51.
  12. Carr R: Neutrophil production and function in newborn infants. Br J Haematol 2000;110:18–28.
  13. Angus DC, Linde-Zwirble WT, Lidicker J, Clermont G, Carcillo J, Pinsky MR: Epidemiology of severe sepsis in the united states: analysis of incidence, outcome, and associated costs of care. Crit Care Med 2001;29:1303–1310.
  14. Stoll BJ, Hansen N, Fanaroff AA, Wright LL, Carlo WA, Ehrenkranz RA, Lemons JA, Donovan EF, Stark AR, Tyson JE, Oh W, Bauer CR, Korones SB, Shankaran S, Laptook AR, Stevenson DK, Papile LA, Poole WK: Changes in pathogens causing early-onset sepsis in very-low-birth-weight infants. N Engl J Med 2002;347:240–247.
  15. Stoll BJ, Hansen NI, Higgins RD, Fanaroff AA, Duara S, Goldberg R, Laptook A, Walsh M, Oh W, Hale E: Very low birth weight preterm infants with early onset neonatal sepsis: the predominance of gram-negative infections continues in the national institute of child health and human development neonatal research network, 2002–2003. Pediatr Infect Dis J 2005;24:635–639.
  16. Hunter CJ, Upperman JS, Ford HR, Camerini V: Understanding the susceptibility of the premature infant to necrotizing enterocolitis (NEC). Pediatr Res 2008;63:117–123.
  17. Speer CP: Pulmonary inflammation and bronchopulmonary dysplasia. J Perinatol 2006;26(suppl 1):S57–S62; discussion S63–S54.
  18. Zentay Z, Sharaf M, Qadir M, Drafta D, Davidson D: Mechanism for dexamethasone inhibition of neutrophil migration upon exposure to lipopolysaccharide in vitro: role of neutrophil interleukin-8 release. Pediatr Res 1999;46:406–410.
  19. Takeuchi O, Akira S: Pattern recognition receptors and inflammation. Cell 2010;140:805–820.
  20. Akira S, Uematsu S, Takeuchi O: Pathogen recognition and innate immunity. Cell 2006;124:783–801.
  21. Takeda K, Akira S: Toll-like receptors in innate immunity. Int Immunol 2005;17:1–14.
  22. Wynn JL, Scumpia PO, Winfield RD, Delano MJ, Kelly-Scumpia K, Barker T, Ungaro R, Levy O, Moldawer LL: Defective innate immunity predisposes murine neonates to poor sepsis outcome but is reversed by TLR agonists. Blood 2008;112:1750–1758.
  23. Lindemann SW, Yost CC, Denis MM, McIntyre TM, Weyrich AS, Zimmerman GA: Neutrophils alter the inflammatory milieu by signal-dependent translation of constitutive messenger RNAs. Proc Natl Acad Sci U S A 2004;101:7076–7081.
  24. Yost CC, Denis MM, Lindemann S, Rubner FJ, Marathe GK, Buerke M, McIntyre TM, Weyrich AS, Zimmerman GA: Activated polymorphonuclear leukocytes rapidly synthesize retinoic acid receptor-alpha: a mechanism for translational control of transcriptional events. J Exp Med 2004;200:671–680.
  25. Caron JE, La Pine TR, Augustine NH, Martins TB, Hill HR: Multiplex analysis of toll-like receptor-stimulated neonatal cytokine response. Neonatology 2010;97:266–273.
  26. Scapini P, Lapinet-Vera JA, Gasperini S, Calzetti F, Bazzoni F, Cassatella MA: The neutrophil as a cellular source of chemokines. Immunol Rev 2000;177:195–203.
  27. Levy O, Suter EE, Miller RL, Wessels MR: Unique efficacy of toll-like receptor 8 agonists in activating human neonatal antigen-presenting cells. Blood 2006;108:1284–1290.
  28. Contrino J, Krause PJ, Slover N, Kreutzer D: Elevated interleukin-1 expression in human neonatal neutrophils. Pediatr Res 1993;34:249–252.
  29. Usmani SS, Kamran S, Harper RG, Wapnir RA, Mehta R: Effect of maternal labor and mode of delivery on polymorphonuclear leukocyte function in healthy neonates. Pediatr Res 1993;33:466–468.
  30. Molloy EJ, O’Neill AJ, Grantham JJ, Sheridan-Pereira M, Fitzpatrick JM, Webb DW, Watson RW: Labor promotes neonatal neutrophil survival and lipopolysaccharide responsiveness. Pediatr Res 2004;56:99–103.
  31. Weinberger B, Vetrano AM, Syed K, Murthy S, Hanna N, Laskin JD, Laskin DL: Influence of labor on neonatal neutrophil apoptosis, and inflammatory activity. Pediatr Res 2007;61:572–577.
  32. Molloy EJ, O’Neill AJ, Doyle BT, Grantham JJ, Taylor CT, Sheridan-Pereira M, Fitzpatrick JM, Webb DW, Watson RW: Effects of heat shock and hypoxia on neonatal neutrophil lipopolysaccharide responses: altered apoptosis, toll-like receptor-4 and CD11B expression compared with adults. Biol Neonate 2006;90:34–39.
  33. Liu Y, Zhu L, Fatheree NY, Liu X, Pacheco SE, Tatevian N, Rhoads JM: Changes in intestinal toll-like receptors and cytokines precede histological injury in a rat model of necrotizing enterocolitis. Am J Physiol Gastrointest Liver Physiol 2009;297:G442–G450.