In Vitro Models for the Study of the Intracellular Activity of Antibiotics.
Frédéric PeyrussonTiep K NguyenJulien M BuyckSandrine LemaireGang WangCristina SeralPaul M TulkensFrançoise Van BambekePublished in: Methods in molecular biology (Clifton, N.J.) (2022)
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.
Keyphrases
- induced apoptosis
- reactive oxygen species
- cell cycle arrest
- stem cells
- escherichia coli
- gene expression
- dna methylation
- signaling pathway
- single cell
- cell death
- quantum dots
- high resolution
- peripheral blood
- living cells
- cell proliferation
- binding protein
- anti inflammatory
- protein protein
- case control
- molecularly imprinted