Human THP-1 monocyte uptake and cellular disposition of 14C-grepafloxacin

• Published in: Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy 2004-02, Vol.10 (1), p.11-18
• Main Authors: Hall, Iris H., Stacy Ward, E., Schwab, Ute E., Rublein, John C., Butts, John D., Ives, Timothy J.
• Format: Article
• Language: English
• Subjects: Carbon Radioisotopes - pharmacokinetics; Cell Line, Tumor - metabolism; Fluoroquinolone; Fluoroquinolones - pharmacokinetics; Grepafloxacin; Humans; Hydrogen-Ion Concentration; Monocytes - metabolism; Piperazines - pharmacokinetics; THP-1 cell; Transport
• ISSN: 1341-321X; 1437-7780
• DOI: 10.1007/s10156-003-0289-8

Uptake of 14C-grepafloxacin into human mononuclear (THP-1) cells was determined at pH 7.4, 6.8, or 5.0 over a 4-log antibiotic concentration. Grepafloxacin was taken up by THP-1 monocytes rapidly by both a passive and an active transport mechanism at pH 7.4. Its uptake was initially linear, with equilibrium being reached after ∼1h. Efflux followed first-order clearance and was complete within 1 h, suggesting no longterm sequestering of the antibiotic occurred. Neither cell number nor serum protein binding appeared to have any effect on antibiotic uptake. High intracellular concentrations were achieved and the ratios of cellular to extracellular antibiotic concentration (IC/EC) were between 529 and 644 at 0.04μg/ml at pH 7.4 and 6.8, suggesting that monocytes may contain sufficient levels of grepafloxacin for affecting bacteriostatic killing. Grepafloxacin disposition within the THP-1 monocytes showed large amounts present in the nucleus and cell sap in stimulated and unstimulated cells, and its presence was evenly distributed throughout the cytosol, nuclei, lysosomes, mitochondria, and ribosomes. After stimulation by zymogen A, Staphylococcus aureus, or Streptococcus pneumoniae, increased amounts of grepafloxacin were found within THP-1 monocytes and isolated phagosome vacuoles. No antibiotic sequestration occurred inside stimulated monocytes, although a sufficient intracellular grepafloxacin concentration was available to kill phagocytized bacteria. Metabolic inhibitors, suppressors of K+/Cl- and Cl- transporters, inhibitors of the phagocytic process, low temperature, and low pH inhibited grepafloxacin uptake by THP-1 monocytes.