The in vivo efficacy of three cell wall-active antibiotics, imipenem, meropenem, and ceftazidime, was compared in mice rendered hypersusceptible to the pathophysiologic effects of lipopolysaccharide by treatment with D-galactosamine. When CF-1 mice were administered Escherichia coli, D-galactosamine, and saline intraperitoneally, an LD50 was achieved at an inoculum of approximately 2 x 10(4) cfu. Administration of antibiotic at 20 mg/kg resulted in significant but widely variable protective efficacy from E. coli lethality among the three antibiotics. At this dose, an approximately 3-fold increase in LD50 was observed with either meropenem or ceftazidime, whereas administration of imipenem resulted in an approximately 8-fold increase in LD50 (P = .0053). When the dose of antibiotic was decreased to 2 mg/kg, neither meropenem nor ceftazidime could provide measurable protection, whereas imipenem was almost fully protective (P < .002). These differences in protective efficacy were also noted with experimental Pseudomonas aeruginosa but not Staphylococcus aureus infection.

Numerous studies have suggested that lipopolysaccharide (LPS), a major component of the cell wall of gram-negative bacteria, is responsible for the initiation of gram-negative septic shock. Previously, others have designed therapeutic regimens to target the biologically active lipid A region of LPS by either neutralization of the biological properties of LPS or enhancement of clearance of this molecule. One such compound capable of neutralizing lipid A is the antibiotic polymyxin B. However, the clinical utility of polymyxin B is limited by its toxicity. We therefore covalently conjugated this antibiotic to the high-molecular-weight polysaccharide dextran 70, resulting in reduced toxicity of polymyxin B but retention of its endotoxin-neutralizing ability. The studies described in this report were designed to test the in vivo efficacy of this compound in an experimental animal model of gram-negative septic shock. Mice were administered graded doses of Escherichia coli or Pseudomonas aeruginosa along with D-galactosamine and the antibiotic imipenem. We had previously determined that antibiotic chemotherapy provides significant protection against E. coli-mediated lethality with smaller doses of bacteria; however, the antibiotic does not provide protection against larger doses of bacteria, but it is effective at killing the bacterial inoculum in vivo. Administration of the polymyxin B-dextran 70 conjugate provided significant protection when given with an antibiotic but was not effective by itself. A requirement for a pretreatment period prior to E. coli challenge was shown to depend upon the bacterial challenge dose. In other studies using this D-galactosamine sensitization model, we demonstrated that the lipid A-specific conjugate had no effect on lethality caused by Staphylococcus aureus or tumor necrosis factor alpha. The results of these studies indicate that this compound is effective in preventing lethal gram-negative septic shock in mice and may be useful as a potential therapeutic agent in humans as well.

Treatment of log phase cultures of Escherichia coli with cell wall active antibiotics results in increased exposure of immunologically reactive lipid A epitopes of lipopolysaccharide (LPS) and release of soluble LPS into culture supernatants. Comparison of the efficacy of two cell wall active antibiotics, ceftazidime, a penicillin-binding protein 3 selective antibiotic, and imipenem, a penicillin-binding protein 2 selective antibiotic, for their relative efficacy in mediating LPS release indicated quantitative but not qualitative differences, with the former antibiotic manifesting a significantly broader range of concentrations at which LPS release could be demonstrated. Comparison of the relative efficacy of these two antibiotics in a mouse bacteraemia model in which animals were made hypersensitive to the lethal effects of endotoxin by treatment with D-galactosamine indicated that the latter antibiotic may provide a greater level of protection. These studies suggest that the release of endotoxin mediated by antibiotic treatment may contribute to the pathogenesis of disease in infectious due to gram-negative organisms.

The objective was to characterize the antibody isotype responses to vaccination with a commercial Staphylococcus aureus bacterin in dairy heifers. Twenty-five Holstein-Friesian dairy heifers were assigned at random to one of two groups, vaccinates (n=14) or controls (n=11). Vaccinates received two 5-ml doses of Lysigin 28 d apart in late gestation. Both groups were challenged with a heterologous serotype 5 strain of Staph. aureus by aseptic intramammary infusion on days 6, 7 and 8 of lactation. Samples for serum antibody isotype (IgG, IgG1, IgG2 and IgM) determinations were taken before each vaccination, immediately prior to challenge (6 d post-calving) and at the end of the study (28 d post-calving). Samples for milk antibody isotype determinations were taken immediately prior to challenge and at the end of the study. Antibody isotype sample-to-positive ratios (S:P ratio=(mean test sample OD-mean within-plate negative control OD)/(mean within-plate positive control OD-mean within-plate negative control OD)) were determined in milk and serum using a series of ELISAs coated with different strains of Staph. aureus belonging to capsular polysaccharide (CP) serotype 5 or 8 or surface polysaccharide (SP) serotype 336. Vaccinates had higher mean serum IgG1 and IgG2 S:P ratios than controls against all three strains of Staph. aureus (P < or = 0.023). Vaccinates had higher mean milk IgG S:P ratios than controls against CP8 and SP336 strains of Staph. aureus (P< or = 0.030). Hence, a humoural immune response to the vaccine was detected in serum and milk, but responses varied according to strain and antibody isotype tested.