Intracellular models of infection to evaluate antibiotic activity

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Presentation transcript:

Intracellular models of infection to evaluate antibiotic activity M. Barcia-Macay, S. Carryn, S. Lemaire, C. Seral, M.P. Mingeot-Leclercq, P.M. Tulkens, F. Van Bambeke Pharmacologie cellulaire et moléculaire Université catholique de Louvain, Brussels, Belgium

Intracellular killing of bacteria by host cell defense mechanisms Carryn et al, Infect Dis Clin North Am. (2003) 17:615-34 Phagosomes Lysosomes Phagolysosomes

Some bacteria can escape host cell defense mechanisms Carryn et al, Infect Dis Clin North Am. (2003) 17:615-34 Early endosomes Phagosomes Phagosomes Inclusions Mycobacterium spp. Chlamydia spp. Salmonella spp. Brucella spp. Lysosomes Cytosol Listeria monocytogenes Shigella flexeneri Endoplasmic reticulum Phagolysosomes Legionella pneumophila Staphylococcus aureus Legionella pneumophila

Setting – up models of intracellular infections L. monocytogenes S. aureus infection of THP-1 macrophages and phagocytosis decomplemented serum; 5 bact./cell serum-opsonized; 4 bact./cell elimination of extracellular bacteria washing washing 1 h with GEN 50 mg/L incubation for 24 h control: + GEN (1 X MIC) antibiotic 1.0 mg/L 0.5 mg/L Cmax

Setting – up models of intracellular infections L. monocytogenes 6 12 18 24 2 4 extra intra time (h) D log CFU from time 0 h S. aureus 6 12 18 24 2 4 extra intra time (h) D log CFU from time 0 h

Testing antibiotics : azithromycin AZM Cmax: 0.4 mg/L MIC L.m.: 0.5 mg/L MIC S.a.: 0.5 mg/L cellular accumulation: 38 X S. aureus extra intra -4 -2 2 D log CFU from time 0 AZM ctr AZM L. monocytogenes poorly active extra- and intra-cellularly !

Testing antibiotics : gentamicin Cmax: 18 mg/L MIC L.m.: 1.0 mg/L MIC S.a.: 0.5 mg/L cellular accumulation: 4.4 X S. aureus extra intra -4 -2 2 D log CFU from time 0 GEN ctr L. monocytogenes highly active extra; static intra-cellularly !

Testing antibiotics : moxifloxacin Cmax: 4 mg/L MIC L.m.: 0.5 mg/L MIC S.a.: 0.06 mg/L MXF cellular accumulation: 7.6 X MXF S. aureus extra intra -4 -2 2 D log CFU from time 0 MXF ctr L. monocytogenes somewhat more active extra- than intra-cellularly !

Testing antibiotics : ampicillin Cmax: 50 mg/L MIC L.m.: 1 mg/L MIC S.a.: 0.06 mg/L AMP cellular accumulation: 1 X AMP S. aureus extra intra -4 -2 2 D log CFU from time 0 AMP ctr L. monocytogenes more active intra- than extra-cellularly against L.m. !

Intracellular activity of antibiotics: what have we learned ? intracellular activity variable according to the localization of the bacteria of the antibiotic intracellular activity most often lower than extracellular activity (exception : ampicillin and L. monocytogenes) intracellular activity not predictible on the basis of MIC accumulation appropriate models are needed !

Why these discrepancies ? Intracellular PK/PD parameters should be taken into account …. Carryn et al, Infect Dis Clin North Am. (2003) 17:615-34

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