Intracellular accumulation, subcellular distribution, and efflux of tilmicosin in chicken phagocytes.
Tilmicosin is a semi-synthetic macrolide antibiotic, currently approved for veterinary use in cattle and swine respiratory disease, and is in development for use in poultry mycoplasma air sacculitis. In order to provide an understanding of clinical efficacy, the in vitro interaction of tilmicosin with three types of chicken phagocytes (MQ-NCSU macrophages, monocyte-macrophages, and heterophils) was evaluated. After incubation with radiolabeled tilmicosin, uptake was determined and expressed as the ratio of the cellular (Cc) to the extracellular (Ce) drug concentration (Cc:Ce). Tilmicosin was avidly accumulated by heterophils (Cc: Ce 138 at 4 h incubation vs 32 and 66, respectively, in MQ-NCSU and monocyte-macrophages) with 61 to 88% localized in the lysosomes. Uptake was dependent on cell viability, temperature, and pH, but was not influenced by metabolic inhibitors. However, phagocytosis of Pasteurella multocida and lipopolysaccharide exposure increased tilmicosin uptake by the chicken phagocytes. Upon removal of extracellular tilmicosin, 50% of the intracellular tilmicosin was effluxed within the first 30 min, but after 4 h of incubation in antibiotic-free medium, 30% remained cell-associated. Opsonized P. multocida significantly enhanced the release of tilmicosin from all three types of chicken phagocytes. Tilmicosin uptake was observed to increase lysosomal enzyme (acid phosphatase, lysozyme, avidin, and beta-glucuronidase) production. Finally, neutrophils were shown to transport and efflux bioactive tilmicosin in a test system measuring both neutrophil chemotaxis under agarose and a bioassay measuring inhibition of bacterial growth in the presence of antibiotic in agar. These in vitro observations of cellular pharmacology suggest a complex interaction between phagocytes and tilmicosin that contribute to clinical efficacy.
Animal Science Research, Elanco Animal Health, Greenfield, Indiana 46140, USA.
B
B
Scorneaux
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TR
Shryock
D000818:Animals; D000900:Anti-Bacterial Agents / Q000097:blood / Q000493:pharmacokinetics*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002645:Chickens; D007700:Kinetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D018942:Macrolides; D008264:Macrophages / Q000502:physiology; D008297:Male; D009000:Monocytes / Q000502:physiology; D016979:Pasteurella multocida; D010431:Pentoxifylline / Q000494:pharmacology; D010586:Phagocytes / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D010587:Phagocytosis; D013347:Subcellular Fractions / Q000378:metabolism; D015645:Tylosin / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics
D000900:Anti-Bacterial Agents; D008070:Lipopolysaccharides; D018942:Macrolides; D010431:Pentoxifylline; C052319:tilmicosin; D015645:Tylosin