Mode of bacterial killing affects the inflammatory response and associated organ dysfunctions in a porcine E. coli intensive care sepsis model

Paul Skorup, Lisa Maudsdotter, Miklós Lipcsey, Anders Larsson, Jan Sjölin, Paul Skorup, Lisa Maudsdotter, Miklós Lipcsey, Anders Larsson, Jan Sjölin

Abstract

Background: Sepsis is often treated with penicillin-binding protein 3 (PBP-3) acting β-lactam antibiotics, such as piperacillin-tazobactam, cefotaxime, and meropenem. They cause considerable bacterial structural changes and have in vitro been associated with an increased inflammatory response. In a clinically relevant large animal sepsis model, our primary aim was to investigate whether bacteria killed by a PBP-3-active antibiotic has a greater effect on the early inflammatory response and organ dysfunction compared with corresponding amounts of live or heat-killed bacteria. A secondary aim was to determine whether the addition of an aminoglycoside could mitigate the cefuroxime-induced response.

Method: Killed or live Escherichia coli were administrated as a 3-h infusion to 16 healthy pigs in a prospective, randomized controlled interventional experimental study. Cefuroxime was chosen as the PBP-3-active antibiotic and tobramycin represented the aminoglycosides. The animals were randomized to receive (I) bacteria killed by cefuroxime, (II) live bacteria, (III) bacteria killed by heat, or (IV) bacteria killed by the combination of cefuroxime and tobramycin. Plasma endotoxin, tumor necrosis factor alpha, interleukin-6, interleukin-10, leukocytes, and organ function were recorded at the start of the experiment and then hourly for 6 h.

Results: Differences in dynamics of concentration over time between the four treatment groups were found for the three cytokines (p < 0.001). Animals receiving cefuroxime-killed bacteria demonstrated higher responses than those receiving live (p < 0.05) or heat-killed bacteria (p < 0.01). The addition of tobramycin reduced the cefuroxime-induced responses (p < 0.001). The cytokine responses were associated with leucocyte activation that was further associated with pulmonary dysfunction and increases in lactate (p < 0.01).

Conclusions: In comparison with live or heat-killed bacteria, bacteria killed by a PBP-3-active antibiotic induced an increased inflammatory response that appears to be associated with deteriorated organ and cellular function. The addition of an aminoglycoside to the PBP-3-active antibiotic reduced that response.

Keywords: Antibiotics; Bacteria; Cytokines; Inflammation; Porcine; Sepsis.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
The experimental protocol. The black bar represents an infusion of live or killed bacteria and the gray bar constitutes antibiotics given to eradicate eventual remaining live bacteria in groups I, III, and IV. White arrows indicate time of sampling for bacterial quantification, cytokine concentrations, and organ function parameters. Black arrows designate time of sampling for endotoxin concentrations
Fig. 2
Fig. 2
Dynamics in concentration of TNF-α, IL-6, and IL-10 in animals during and after administration of killed or live E. coli in the following treatment groups (n = 4 in each group): (I) bacteria killed by cefuroxime alone, (II) live bacteria, (III) bacteria killed by heat, and (IV) bacteria killed by the combination of cefuroxime and tobramycin. Data are shown as mean ± SE. p values are the results of the group-by-time interaction in the repeated-measures ANOVA after considering all time points

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