Abstract

Bacterial neonatal meningitis results in high mortality and morbidity rates for those affected. Although improvements in diagnosis and treatment have led to a decline in mortality rates, morbidity rates have remained relatively unchanged. Bacterial resistance to antibiotics in this clinical setting further underlines the need for developing other technologies, such as phage therapy. We exploited an in vitro phage therapy model for studying bacterial neonatal meningitis based on Escherichia coli (E. coli) EV36, bacteriophage (phage) K1F and human cerebral microvascular endothelial cells (hCMECs). We show that phage K1F is phagocytosed and degraded by constitutive- and PAMP-dependent LC3-assisted phagocytosis and does not induce expression of inflammatory cytokines TNFα, IL-6, IL-8 or IFNβ. Additionally, we observed that phage K1F temporarily decreases the barrier resistance of hCMEC cultures, a property that influences the barrier permeability, which could facilitate the transition of immune cells across the endothelial vessel in vivo. Collectively, we demonstrate that phage K1F can infect intracellular E. coli EV36 within hCMECs without themselves eliciting an inflammatory or defensive response. This study illustrates the potential of phage therapy targeting infections such as bacterial neonatal meningitis and is an important step for the continued development of phage therapy targeting antibiotic-resistant bacterial infections generally.

Highlights

  • Improvements in diagnosis and treatment of bacterial neonatal meningitis have seen a gradual decline in mortality rates in recent decades, while long term post-infection morbidity rates have remained relatively unchanged[3]

  • We present a robust in vitro phage therapy model system of neonatal bacterial meningitis based on E. coli EV36, phage K1F and human cerebral microvascular endothelial cells (hCMECs) allowing for a wide range of cellular and molecular analyses

  • Infection rates of hCMECs by E. coli EV36-RFP were determined by confocal microscopy and flow cytometry (Fig. 1)

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Summary

Introduction

Improvements in diagnosis and treatment of bacterial neonatal meningitis have seen a gradual decline in mortality rates in recent decades, while long term post-infection morbidity rates have remained relatively unchanged[3]. Phages are present on all body surfaces that are in direct contact with the exterior environment, including the skin, urogenital tract, oral cavity, gut and lungs[15], in addition to the blood[16]. While their presence in body niches allows them to exert selective pressure on their bacterial hosts and to modulate the human microbiome, their presence in the blood allows for direct interaction with mammalian immune cells and the potential for induction of innate and adaptive immune responses[17]. This decrease in focal adhesion influences the endothelial barrier function by increasing the barrier permeability and this might represent a mechanism allowing for the transition of immune cells across the endothelial vessel

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