title: Chemiluminescence of human polymorphonuclear leucocytes after stimulation with whole cells and cell-wall components of Staphylococcus epidermidis.
abstract: The purpose of this study was to define cell-wall components of Staphylococcus epidermidis responsible for activation of human polymorphonuclear leucocytes (PMNL). Metabolic activation of PMNL was determined by chemiluminescence (CL). Purified peptidoglycan (PG) induced a concentration-dependent metabolic burst in PMNL. The minimal concentration needed for CL induction was 1 microgram/ml. Comparison between different S. epidermidis strains showed variation in the capacity to induce CL in PMNL. Purified PG induced a higher CL response in PMNL than its intact parent strain; this effect was found in all S. epidermidis strains. Lipoteichoic acid (LTA), PG stem peptide and muramyldipeptide (MDP) did not induce CL; teichoic acid induced a CL response only at very high concentrations. No differences in CL inducing capacity were found between PG, crude cell walls, and purified cell walls of S. epidermidis. Sonication of PG strongly diminished CL-inducing capacity. PG treatment with mutanolysin immediately resulted in decreased CL-inducing capacity. Treatment of PG with S. aureus lytic enzyme (SALE) 10 micrograms/ml for up to 15 min enhanced the CL response to PMNL; a similar increase in CL was induced by PG treated with SALE at 1 microgram/ml for up to 120 min. Beyond these times, a continuous decrease in PG-induced CL was observed. In conclusion, PG was found to be the major cell-wall component of S. epidermidis involved in CL induction. Moreover, a minimal fragment size or a specific tertiary structure of PG, or both, is required for metabolic activation of PMNL.
authors.affiliation: Eijkman-Winkler Laboratory of Medical Microbiology, University of Utrecht, The Netherlands.
authors.forename: L
authors.initials: L
authors.lastname: Martínez-Martínez
authors.forename: C P
authors.initials: CP
authors.lastname: Timmerman
authors.forename: A
authors.initials: A
authors.lastname: Fleer
authors.forename: J
authors.initials: J
authors.lastname: Verhoef
mesh_terms: D000328:Adult; D002473:Cell Wall / Q000737:chemistry / Q000276:immunology; D004306:Dose-Response Relationship, Immunologic; D010450:Endopeptidases / Q000378:metabolism; D006801:Humans; D008070:Lipopolysaccharides / Q000276:immunology / Q000302:isolation & purification; D008163:Luminescent Measurements; D009504:Neutrophils / Q000276:immunology*; D009895:Opsonin Proteins / Q000276:immunology; D010457:Peptidoglycan / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D016897:Respiratory Burst; D013010:Sonication; D013211:Staphylococcus aureus / Q000201:enzymology; D013212:Staphylococcus epidermidis / Q000737:chemistry / Q000276:immunology* / Q000648:ultrastructure; D013682:Teichoic Acids / Q000276:immunology / Q000302:isolation & purification
chemical_list: D008070:Lipopolysaccharides; D009895:Opsonin Proteins; D010457:Peptidoglycan; D013682:Teichoic Acids; C009900:lipoteichoic acid; D010450:Endopeptidases; C021816:mutanolysin