Effects of neutrophils on cefazolin activity and penicillin-binding proteins in Staphylococcus aureus abscesses.
Bacteria survive within abscesses despite antimicrobial therapy, usually necessitating drainage. Our previous work showed that bacterial killing is diminished within the neutrophils of animals with abscesses. To further assess the role of neutrophils in Staphylococcus aureus survival and the poor activities of beta-lactams in abscesses, tissue cage abscess-bearing rats were given polymorphonuclear leukocyte (PMN)-depleting antibody prior to and several times following inoculation of the tissue cages with S. aureus. Cefazolin (300 mg/kg of body weight/day) was administered to all animals in appropriately divided doses. After 7 days of antimicrobial therapy, the 17 animals that received anti-PMN serum had significantly fewer abscess neutrophils than the 18 controls and fewer abscess bacteria (5.55 versus 3.79 log(10) CFU/ml [P = 0.04]) than the 18 controls. The data were consistent with the premise that cefazolin is more effective in abscesses depleted of neutrophils. To investigate further, S. aureus was incubated with rat peritoneal neutrophils; and bacterial cell membrane proteins were isolated, labeled with biotinylated ampicillin, separated by electrophoresis, blotted onto nitrocellulose, and stained for biotin reactivity. PBP 2 expression was consistently and significantly decreased after a brief, nonkilling PMN exposure. These experiments showed that PMN depletion enhanced the activity of cefazolin in the abscess milieu. Furthermore, altered bacterial cell wall cefazolin targets may be the mechanism by which the PMN diminishes antimicrobial activity, suggesting the importance of the staphylococcus-PMN interaction in the outcome of established infections.
Bamberger
David M
DM
Section of Infectious Diseases, Department of Medicine, University of Missouri--Kansas City, Kansas City, Missouri 64108, USA. bambergerd@umkc.edu
Herndon
Betty L
BL
Fitch
Jeffrey
J
Florkowski
Aaron
A
Parkhurst
Vera
V
D000038:Abscess / Q000276:immunology / Q000382:microbiology*; D000667:Ampicillin / Q000494:pharmacology; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins; D001710:Biotin; D002352:Carrier Proteins / Q000378:metabolism*; D002437:Cefazolin / Q000494:pharmacology*; D005456:Fluorescent Dyes; D006602:Hexosyltransferases; D018655:Lymphocyte Count; D008297:Male; D002267:Muramoylpentapeptide Carboxypeptidase / Q000378:metabolism*; D009503:Neutropenia / Q000382:microbiology; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D046915:Penicillin-Binding Proteins; D010458:Peptidyl Transferases; D010587:Phagocytosis / Q000187:drug effects; D051381:Rats; D013203:Staphylococcal Infections / Q000276:immunology / Q000382:microbiology*
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D002352:Carrier Proteins; D005456:Fluorescent Dyes; D046915:Penicillin-Binding Proteins; D001710:Biotin; D000667:Ampicillin; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase; D002437:Cefazolin