Killing of an encapsulated strain of Escherichia coli by human serum.
Changes in cell viability and in factors affecting metabolic integrity were examined after exposure of Escherichia coli LP1092 to human serum. Antibody-dependent classical pathway activity accounted for the rapid killing of strain LP1092 by complement. Removal of serum lysozyme by bentonite absorption or by neutralization with anti-human lysozyme immunoglobulin G resulted in a reduction in the rate of killing; optimal activity could be restored by the addition of physiological amounts of egg-white lysozyme. The pattern of 86Rb+ and alkaline phosphatase release obtained after serum treatment did not support the view that complement simultaneously disrupts cytoplasmic and outer membrane integrity. Macromolecular synthesis was affected late in the reaction sequence; complete inhibition of precursor incorporation into RNA, DNA, and protein occurred only after almost total loss of bacterial colony-forming ability. Addition of chloramphenicol, an inhibitor of protein synthesis, to the bactericidal system resulted in a marked reduction in the rate of serum killing. Killing was completely inhibited by an inhibitor (KCN) and an uncoupler (2,4-dinitrophenol) of oxidative phosphorylation. Exposure of LP1092 cells to serum was followed by a rapid and large increase in intracellular ATP levels; ATP synthesis did not occur when bacteria were exposed to dialyzed serum, which killed LP1092 cells at a much reduced rate. Addition of glucose or serum ultrafiltrate to dialyzed serum restored optimal bactericidal activity. We suggest that optimal killing of gram-negative bacteria is an energy-dependent process requiring an input of bacterially generated ATP.
D000255:Adenosine Triphosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000096:biosynthesis; D001770:Blood Bactericidal Activity; D002701:Chloramphenicol / Q000494:pharmacology; D003165:Complement System Proteins / Q000276:immunology; D004269:DNA, Bacterial / Q000096:biosynthesis; D004926:Escherichia coli / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D009113:Muramidase / Q000494:pharmacology; D011190:Potassium Cyanide / Q000494:pharmacology; D012329:RNA, Bacterial / Q000096:biosynthesis
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D002701:Chloramphenicol; D000255:Adenosine Triphosphate; D003165:Complement System Proteins; D009113:Muramidase; D011190:Potassium Cyanide