Department of Medical Microbiology, Amsterdam UMC, University of Amsterdam, Amsterdam Infection and Immunity Institute, Amsterdam, The Netherlands.
Methods Mol Biol. 2022;2451:671-689. doi: 10.1007/978-1-0716-2099-1_35.
Pathogens such as Staphylococcus aureus are able to survive in many types of host cells including phagocytes such as neutrophils and macrophages, thereby resulting in intracellular infections. Treatment of intracellular infections by conventional antimicrobials (e.g., antibiotics) is often ineffective due to low intracellular efficacy of the drugs. Thus, novel techniques which can enhance the activity of antimicrobials within cells are highly demanded. Our recent studies have shown that photochemical internalization (PCI) is a promising approach for improving the efficacy of antibiotics such as gentamicin against intracellular staphylococcal infection. In this chapter, we describe the protocols aiming to study the potential of PCI-antibiotic treatment for intracellular infections in vitro and in vivo using a RAW 264.7 cell infection model and a zebrafish embryo infection model. Proof of concept of this approach is demonstrated. The protocols are expected to prompt further development of PCI-antimicrobial based novel therapies for clinically challenging infectious diseases associated with intracellular survival of pathogens.
金黄色葡萄球菌等病原体能够在包括中性粒细胞和巨噬细胞等吞噬细胞在内的多种宿主细胞中存活,从而导致细胞内感染。由于药物在细胞内的疗效较低,传统的抗菌药物(例如抗生素)治疗细胞内感染往往效果不佳。因此,人们迫切需要能够提高抗菌药物在细胞内活性的新技术。我们最近的研究表明,光化学内化(PCI)是提高庆大霉素等抗生素对细胞内金黄色葡萄球菌感染疗效的一种很有前途的方法。在本章中,我们描述了使用 RAW 264.7 细胞感染模型和斑马鱼胚胎感染模型,研究 PCI-抗生素治疗细胞内感染的体外和体内潜力的方案。该方法的概念验证已得到证明。预计这些方案将促进基于 PCI-抗菌药物的新型疗法的进一步发展,以治疗与病原体在细胞内生存有关的具有临床挑战性的传染病。