title: Dynamics of human complement-mediated killing of Klebsiella pneumoniae.
abstract: With an in vitro system that used a luminescent strain of Klebsiella pneumoniae to assess bacterial metabolic activity in near-real-time, we investigated the dynamics of complement-mediated attack in healthy individuals and in patients presenting to the emergency department with community-acquired severe sepsis. A novel mathematical/statistical model was developed to simplify light output trajectories over time into two fitted parameters, the rate of complement activation and the delay from activation to the onset of killing. Using Factor B-depleted serum, the alternative pathway was found to be the primary bactericidal effector: In the absence of B, C3 opsonization as measured by flow cytometry did not progress and bacteria proliferated near exponentially. Defects in bacterial killing were easily demonstrable in patients with severe sepsis compared with healthy volunteers. In most patients with sepsis, the rate of activation was higher than in normal subjects but was associated with a prolonged delay between activation and bacterial killing (P < 0.05 for both). Theoretical modeling suggested that this combination of accentuated but delayed function should allow successful bacterial killing but with significantly greater complement activation. The use of luminescent bacteria allowed for the development of a novel and powerful tool for assessing complement immunology for the purposes of mechanistic study and patient evaluation.
authors.affiliation: Department of Emergency Medicine, University of Michigan, Ann Arbor, Michigan 48109-3300, USA.
authors.forename: Christina M
authors.initials: CM
authors.lastname: Nypaver
authors.forename: Margaret M
authors.initials: MM
authors.lastname: Thornton
authors.forename: Suellen M
authors.initials: SM
authors.lastname: Yin
authors.forename: David O
authors.initials: DO
authors.lastname: Bracho
authors.forename: Patrick W
authors.initials: PW
authors.lastname: Nelson
authors.forename: Alan E
authors.initials: AE
authors.lastname: Jones
authors.forename: David M
authors.initials: DM
authors.lastname: Bortz
authors.forename: John G
authors.initials: JG
authors.lastname: Younger
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D003176:Complement C3 / Q000276:immunology; D003165:Complement System Proteins / Q000276:immunology*; D006262:Health; D006801:Humans; D007711:Klebsiella pneumoniae / Q000166:cytology* / Q000187:drug effects / Q000276:immunology*; D008163:Luminescent Measurements; D050296:Microbial Viability / Q000187:drug effects / Q000276:immunology*; D009895:Opsonin Proteins / Q000276:immunology; D018805:Sepsis / Q000276:immunology / Q000382:microbiology; D044967:Serum; D013997:Time Factors
chemical_list: D000900:Anti-Bacterial Agents; D003176:Complement C3; D009895:Opsonin Proteins; D003165:Complement System Proteins