Differences in mode of action of (β-lactam antibiotics influence morphology,LPS release and in vivo antibiotic efficacy

Abstract

Antibiotic mediated release of endotoxin (lipopolysaccharide, LPS) from Gram-negative bacteria is implicated in septic shock. The β-lactam (cell wall active) class, in particular, has been deemed responsible for release of greater quantities of LPS than other classes of antibiotics. However, it is becoming increasingly recognized that variations in the ability to liberate free LPS exist within the β-lactam subclasses. Until recently, LPS-release studies have primarily included the cephalosporin, monobactam, and penicillin β-lactams, but not the carbapenem subclass. We document here that carbapenems significantly liberate less LPS than other β-lactam subclasses, and that disparity in LPS release also occurs within the carbapenems as well (i.e. imipenem vs meropenem). The propensity to release LPS correlates with the cumulative PBP binding affinity of each β-lactam antibiotic regardless of subclass. A direct correlation has been established between antibiotic concentration, frequency of antibiotic exposure, differential LPS release, PBP specificity (morphology, rate of bacterial lysis or cell mass) and antibiotic efficacy in two mouse models of infection. In the first, differences in the pathogenic (endotoxic) potential of potently biologically active smooth LPS (S-LPS) and poorly biologically active rough chemotype LPS (R-LPS) expressing bacteria underscores the in vivo importance of endotoxin in mediating lethality following antibiotic chemotherapy. Endotoxin is, however, not innately toxic and thus its lethal effects are mediated through LPS stimulation of host cells (i.e. macrophages) to produce deleterious levels of cytokines. In the second experimental model, therefore, the toxic effects of antibiotic-liberated endotoxin have been abrogated by pretreatment of mice with carrageenan (CGN) prior to challenge with endotoxin or Gram-negative bacteria. CGN treatment eliminates or markedly reduces the numbers of LPS-responsive mediator cells thus affecting mortality. In both animal models, an in vivo role for LPS is demonstrated through differential changes in antibiotic efficacy (ED ₅₀) in response to differential effects of LPS via modulation of the endotoxic sensitivity/responsiveness of the host. We conclude that antibiotic class, concentration, dosing intervals (timing) and perhaps, method of treatment (i.e. bolus vs infusion) may each be important in the survival of experimental animals severely infected with Gram-negative bacteria.