Role of complement and CD14 in meconium-induced cytokine formation.
OBJECTIVE
Meconium aspiration syndrome has a complex, poorly defined pathophysiology. Meconium is a potent activator of complement in vitro and in vivo; the latter is associated with a systemic inflammatory response. The complement system and Toll-like receptors are 2 important upstream components of the innate immune system that act partly independently in the inflammatory network. The aim of this study was to investigate the relative role of complement and CD14 in meconium-induced cytokine production.


METHODS
Human adult (n = 6) and cord whole blood (n = 6) anticoagulated with lepirudin was collected and distributed into tubes that contained inhibitory antibodies (anti-CD14, anti-C2, anti-factor D, or combinations thereof). The tubes were preincubated for 5 minutes before addition of meconium or buffer and then incubated for 4 hours at 37 degrees C. Complement activation was measured by quantification of the terminal sC5b-9 complement complex by enzyme-linked immunosorbent assay. A panel of 27 inflammatory mediators (cytokines, chemokines, and growth factors) was measured by using multiplex technology.


RESULTS
Fourteen of the 27 mediators measured were induced by meconium both in cord and adult blood. In cord blood, 2 additional chemokines were induced and the inflammatory response was, in general, more potent. Blocking of complement or CD14 differentially reduced the formation of most mediators, anti-CD14 being more effective. Notably, the combined inhibition of complement and CD14 almost completely abolished meconium-induced formation of the cytokines and the chemokines and markedly reduced the formation of growth factors. The endogenous lipopolysaccharide content of meconium could not explain the CD14-mediated response.


CONCLUSIONS
Meconium-induced triggering of the cytokine network is differentially mediated by complement and CD14. A combined inhibition of these effector mechanisms may be an alternative approach to reduce the inflammatory reaction in meconium aspiration syndrome.
Salvesen
Bodil
B
Institute of Immunology and Department of Pediatric Research, University of Oslo and Rikshospitalet University Hospital, Oslo, Norway. bodil.salvesen@medisin.uio.no
Fung
Michael
M
Saugstad
Ola D
OD
Mollnes
Tom E
TE
D000328:Adult; D003167:Complement Activation; D003165:Complement System Proteins / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism*; D005260:Female; D005312:Fetal Blood / Q000276:immunology*; D006801:Humans; D007113:Immunity, Innate / Q000502:physiology; D066298:In Vitro Techniques; D007231:Infant, Newborn; D018836:Inflammation Mediators / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000378:metabolism*; D008297:Male; D008470:Meconium / Q000276:immunology*; D008471:Meconium Aspiration Syndrome / Q000276:immunology / Q000503:physiopathology; D011247:Pregnancy; D011336:Probability; D012680:Sensitivity and Specificity; D018709:Statistics, Nonparametric
D016207:Cytokines; D018836:Inflammation Mediators; D018950:Lipopolysaccharide Receptors; D003165:Complement System Proteins