Pharmacologie Cellulaire et Moléculaire, Louvain Drug Research Institute, Université catholique de Louvain, Brussels, Belgium.
INSERM U1070 "Pharmacology of Anti-infective Agents", Université de Poitiers, Poitiers, France.
Methods Mol Biol. 2021;2357:239-251. doi: 10.1007/978-1-0716-1621-5_16.
Intracellular bacteria are poorly responsive to antibiotic treatment. Pharmacological studies are thus needed to determine the antibiotics which are the most potent or effective against intracellular bacteria as well as to explore the reasons for poor bacterial responsiveness. An in vitro pharmacodynamic model is described, consisting of (1) phagocytosis of preopsonized bacteria by eukaryotic cells, (2) elimination of noninternalized bacteria with gentamicin, (3) incubation of infected cells with antibiotics, and (4) determination of surviving bacteria by viable cell counting and normalization of the counts based on sample protein content. The use of strains expressing fluorescent proteins under the control of an inducible promoter allows to follow intracellular bacterial division at the individual level and therefore to monitor bacterial persisters that do not multiply anymore.
胞内细菌对抗生素治疗反应不佳。因此，需要进行药理学研究，以确定针对胞内细菌最有效或最有效的抗生素，并探讨细菌反应不佳的原因。本文描述了一种体外药效动力学模型，包括（1）真核细胞吞噬预调理细菌，（2）用庆大霉素消除未内化的细菌，（3）用抗生素孵育感染细胞，以及（4）通过活菌计数和基于样品蛋白含量的计数归一化来确定存活细菌。使用表达受诱导启动子控制的荧光蛋白的菌株，可以在个体水平上跟踪胞内细菌的分裂，从而监测不再繁殖的细菌持久体。
