Abstract
Long-acting nanoformulated antiretroviral therapy (nanoART) induces a range of innate immune migratory, phagocytic and secretory cell functions that perpetuate drug depots. While recycling endosomes serve as the macrophage subcellular depots, little is known of the dynamics of nanoART-cell interactions. To this end, we assessed temporal leukocyte responses, drug uptake and distribution following both intraperitoneal and intramuscular injection of nanoformulated atazanavir (nanoATV). Local inflammatory responses heralded drug distribution to peritoneal cell populations, regional lymph nodes, spleen and liver. This proceeded for three days in male Balb/c mice. NanoATV-induced changes in myeloid populations were assessed by fluorescence-activated cell sorting (FACS) with CD45, CD3, CD11b, F4/80, and GR-1 antibodies. The localization of nanoATV within leukocyte cell subsets was determined by confocal microscopy. Combined FACS and ultra-performance liquid chromatography tandem mass-spectrometry assays determined nanoATV carriages by cell-based vehicles. A robust granulocyte, but not peritoneal macrophage nanoATV response paralleled zymosan A treatment. ATV levels were highest at sites of injection in peritoneal or muscle macrophages, dependent on the injection site. The spleen and liver served as nanoATV tissue depots while drug levels in lymph nodes were higher than those recorded in plasma. Dual polymer and cell labeling demonstrated a nearly exclusive drug reservoir in macrophages within the liver and spleen. Overall, nanoART induces innate immune responses coincident with rapid tissue macrophage distribution. Taken together, these works provide avenues for therapeutic development designed towards chemical eradication of human immunodeficiency viral infection.
Highlights
Human immunodeficiency virus (HIV) therapeutics have consistently evolved over the past three decades as newer antiretroviral (ARV) medicines have come on-line and have demonstrated improved bioavailability, antiviral responses, ease of administration, and reducedPLOS ONE | DOI:10.1371/journal.pone.0145966 December 30, 2015NanoART Macrophage Targeting and Tissue Drug Depots
To assess the temporal and differential cellular responses following treatment, we examined the cellular environment of the peritoneum using fluorescence-activated cell sorting (FACS) for granulocytes (CD11bhigh, GR-1high), mature macrophages (CD11bhigh, F4/80 positive cells), immature macrophages (CD11bmed, GR-1 negative), and an unstained cell population (CD11b negative, GR-1 negative) [28, 31,32,33,34,35]
We examined cellular responses over a 72 h period after IP injection with P188 or zymosan A and compared these findings to responses elicited by 100 mg/kg of nanoformulated atazanavir (nanoATV) (Fig 1)
Summary
The goals of the current study were to uncover the cellular and tissue drug depots of nanoART and their carrying capacities in vivo; a summary of the experimental design is provided in Fig 8
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