title: Hyperglycemic conditions inhibit C3-mediated immunologic control of Staphylococcus aureus.
abstract: BACKGROUND
  Diabetic patients are at increased risk for bacterial infections; these studies provide new insight into the role of the host defense complement system in controlling bacterial pathogens in hyperglycemic environments.
  
  
  METHODS
  The interactions of complement C3 with bacteria in elevated glucose were assayed for complement activation to opsonic forms, phagocytosis and bacterial killing. C3 was analyzed in euglycemic and hyperglycemic conditions by mass spectrometry to measure glycation and structural differences.
  
  
  RESULTS
  Elevated glucose inhibited S. aureus activation of C3 and deposition of C3b and iC3b on the bacterial surface. S. aureus-generated C5a and serum-mediated phagocytosis by neutrophils were both decreased in elevated glucose conditions. Interestingly, elevated glucose increased the binding of unactivated C3 to S. aureus, which was reversible on return to normal glucose concentrations. In a model of polymicrobial infection, S. aureus in elevated glucose conditions depleted C3 from serum resulting in decreased complement-mediated killing of E. coli. To investigate the effect of differing glucose concentration on C3 structure and glycation, purified C3 incubated with varying glucose concentrations was analyzed by mass spectrometry. Glycation was limited to the same three lysine residues in both euglycemic and hyperglycemic conditions over one hour, thus glycation could not account for observed changes between glucose conditions. However, surface labeling of C3 with sulfo-NHS-biotin showed significant changes in the surface availability of seven lysine residues in response to increasing glucose concentrations. These results suggest that the tertiary structure of C3 changes in response to hyperglycemic conditions leading to an altered interaction of C3 with bacterial pathogens.
  
  
  CONCLUSIONS
  These results demonstrate that hyperglycemic conditions inhibit C3-mediated complement effectors important in the immunological control of S. aureus. Mass spectrometric analysis reveals that the glycation state of C3 is the same regardless of glucose concentration over a one-hour time period. However, in conditions of elevated glucose C3 appears to undergo structural changes.
authors.affiliation: Department of Pediatrics, Eastern Virginia Medical School, Norfolk, VA, USA.
authors.forename: Pamela S
authors.initials: PS
authors.lastname: Hair
authors.forename: Charlene G
authors.initials: CG
authors.lastname: Echague
authors.forename: Reuben D
authors.initials: RD
authors.lastname: Rohn
authors.forename: Neel K
authors.initials: NK
authors.lastname: Krishna
authors.forename: Julius O
authors.initials: JO
authors.lastname: Nyalwidhe
authors.forename: Kenji M
authors.initials: KM
authors.lastname: Cunnion
mesh_terms: D000595:Amino Acid Sequence; D003176:Complement C3 / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D050577:Complement C3-C5 Convertases / Q000378:metabolism; D003602:Cytotoxicity, Immunologic / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D006943:Hyperglycemia / Q000276:immunology* / Q000382:microbiology*; D013058:Mass Spectrometry; D050296:Microbial Viability / Q000187:drug effects; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009895:Opsonin Proteins / Q000276:immunology; D010587:Phagocytosis / Q000187:drug effects / Q000276:immunology; D011485:Protein Binding / Q000187:drug effects; D013203:Staphylococcal Infections / Q000276:immunology / Q000382:microbiology; D013211:Staphylococcus aureus / Q000187:drug effects / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D003176:Complement C3; D009895:Opsonin Proteins; D050577:Complement C3-C5 Convertases; D005947:Glucose