Limits...
Cellular Responses and Tissue Depots for Nanoformulated Antiretroviral Therapy.

Martinez-Skinner AL, Araínga MA, Puligujja P, Palandri DL, Baldridge HM, Edagwa BJ, McMillan JM, Mosley RL, Gendelman HE - PLoS ONE (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE, 68198-5880, United States of America.

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.

Show MeSH

Related in: MedlinePlus

Differential cell counts in peritoneal lavage.Following intraperitoneal injection of zymosan A, nanoATV, or P188 (control), cells were collected by peritoneal lavage at 1, 4, 12, 24, 48, and 72 h. Collected cells were stained with antibodies directed against GR-1, CD11b, and F4/80 and sorted by FACS. Mean total cell numbers per mouse ± SEM were calculated from 4–8 mice/treatment group/time point.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4696780&req=5

pone.0145966.g001: Differential cell counts in peritoneal lavage.Following intraperitoneal injection of zymosan A, nanoATV, or P188 (control), cells were collected by peritoneal lavage at 1, 4, 12, 24, 48, and 72 h. Collected cells were stained with antibodies directed against GR-1, CD11b, and F4/80 and sorted by FACS. Mean total cell numbers per mouse ± SEM were calculated from 4–8 mice/treatment group/time point.

Mentions: We posited that nanoART elicits a similar activation state of macrophages in vivo as observed in cultures of human monocyte-derived macrophages without substantial inflammatory responses [30]. To test this hypothesis, we adapted a peritonitis model to quantitatively assess the biological and cellular responses in vivo [31, 32]. To assess the temporal and differential cellular responses following treatment, we examined the cellular environment of the peritoneum using FACS for granulocytes (CD11bhigh, GR-1high), mature macrophages (CD11bhigh, F4/80 positive cells), immature macrophages (differentiating cells) (CD11bmed, GR-1 negative), and an unstained cell population (CD11b negative, GR-1 negative) [28, 31–35]. We examined cellular responses over a 72 h period after IP injection with P188 (the polymer used in nanoATV preparation) or zymosan A (a known inflammatory reagent) and compared these findings to responses elicited by 100 mg/kg of nanoATV (Fig 1).


Cellular Responses and Tissue Depots for Nanoformulated Antiretroviral Therapy.

Martinez-Skinner AL, Araínga MA, Puligujja P, Palandri DL, Baldridge HM, Edagwa BJ, McMillan JM, Mosley RL, Gendelman HE - PLoS ONE (2015)

Differential cell counts in peritoneal lavage.Following intraperitoneal injection of zymosan A, nanoATV, or P188 (control), cells were collected by peritoneal lavage at 1, 4, 12, 24, 48, and 72 h. Collected cells were stained with antibodies directed against GR-1, CD11b, and F4/80 and sorted by FACS. Mean total cell numbers per mouse ± SEM were calculated from 4–8 mice/treatment group/time point.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4696780&req=5

pone.0145966.g001: Differential cell counts in peritoneal lavage.Following intraperitoneal injection of zymosan A, nanoATV, or P188 (control), cells were collected by peritoneal lavage at 1, 4, 12, 24, 48, and 72 h. Collected cells were stained with antibodies directed against GR-1, CD11b, and F4/80 and sorted by FACS. Mean total cell numbers per mouse ± SEM were calculated from 4–8 mice/treatment group/time point.
Mentions: We posited that nanoART elicits a similar activation state of macrophages in vivo as observed in cultures of human monocyte-derived macrophages without substantial inflammatory responses [30]. To test this hypothesis, we adapted a peritonitis model to quantitatively assess the biological and cellular responses in vivo [31, 32]. To assess the temporal and differential cellular responses following treatment, we examined the cellular environment of the peritoneum using FACS for granulocytes (CD11bhigh, GR-1high), mature macrophages (CD11bhigh, F4/80 positive cells), immature macrophages (differentiating cells) (CD11bmed, GR-1 negative), and an unstained cell population (CD11b negative, GR-1 negative) [28, 31–35]. We examined cellular responses over a 72 h period after IP injection with P188 (the polymer used in nanoATV preparation) or zymosan A (a known inflammatory reagent) and compared these findings to responses elicited by 100 mg/kg of nanoATV (Fig 1).

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE, 68198-5880, United States of America.

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.

Show MeSH
Related in: MedlinePlus