Limits...
Opposing regulation of endolysosomal pathways by long-acting nanoformulated antiretroviral therapy and HIV-1 in human macrophages.

Araínga M, Guo D, Wiederin J, Ciborowski P, McMillan J, Gendelman HE - Retrovirology (2015)

Bottom Line: NanoART and native ATV treated uninfected cells showed limited effects.The data was confirmed by Western blot.DAVID and KEGG bioinformatics analyses of proteomic data showed relationships between secretory, mobility and phagocytic cell functions and virus and particle trafficking.

View Article: PubMed Central - PubMed

Affiliation: Departments of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, 985880 Nebraska Medical Center, Omaha 68198-5880, NE, USA. hegendel@unmc.edu.

ABSTRACT

Background: Long-acting nanoformulated antiretroviral therapy (nanoART) is designed to improve patient regimen adherence, reduce systemic drug toxicities, and facilitate clearance of human immunodeficiency virus type one (HIV-1) infection. While nanoART establishes drug depots within recycling and late monocyte-macrophage endosomes, whether or not this provides a strategic advantage towards viral elimination has not been elucidated.

Results: We applied quantitative SWATH-MS proteomics and cell profiling to nanoparticle atazanavir (nanoATV)-treated and HIV-1 infected human monocyte-derived macrophages (MDM). Native ATV and uninfected cells served as controls. Both HIV-1 and nanoATV engaged endolysosomal trafficking for assembly and depot formation, respectively. Notably, the pathways were deregulated in opposing manners by the virus and the nanoATV, likely by viral clearance. Paired-sample z-scores, of the proteomic data sets, showed up- and down- regulation of Rab-linked endolysosomal proteins. NanoART and native ATV treated uninfected cells showed limited effects. The data was confirmed by Western blot. DAVID and KEGG bioinformatics analyses of proteomic data showed relationships between secretory, mobility and phagocytic cell functions and virus and particle trafficking.

Conclusions: We posit that modulation of endolysosomal pathways by antiretroviral nanoparticles provides a strategic path to combat HIV infection.

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Related in: MedlinePlus

NanoATV treatment affects HIV-1 reverse transcriptase (RT) activity. HIV-1 RT activity was determined in treated (native ATV or nanoATV) MDM followed by HIV-1 infection at days 0, 5 or 10. HIV-1 infected cells without any treatment served as a positive control for RT activity. All samples were collected after 7 days of viral infection. Results shown are the mean of 5 replicates.
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Fig4: NanoATV treatment affects HIV-1 reverse transcriptase (RT) activity. HIV-1 RT activity was determined in treated (native ATV or nanoATV) MDM followed by HIV-1 infection at days 0, 5 or 10. HIV-1 infected cells without any treatment served as a positive control for RT activity. All samples were collected after 7 days of viral infection. Results shown are the mean of 5 replicates.

Mentions: To confirm the antiretroviral activity of nanoATV treatment HIV-1 reverse transcriptase (RT) activity was determined in HIV-1-infected human monocyte-derived macrophages (MDM) treated with either native- or nanoATV. Cells were treated with 10, 100 or 250 μM of native- or nanoATV for 16 hours. At this time the medium was removed, cells were washed 3 times with phosphate buffered saline and fresh medium without drug was added prior to HIV-1ADA challenge at a multiplicity of infection (MOI) of 0.1 at days 0, 5 and 10 after treatment. Infected cells were cultured for an additional 7 days and RT activity in the culture medium was determined. Significant differences were found between cells treated with native- or nanoATV. For native ATV treated cells, RT activity was suppressed only in the day 0 infection group, at all treatment concentrations. At 10 and 100 μM little antiviral suppression was observed in the days 5 and 10 infection groups. In contrast, for cells treated with nanoATV, RT activity was suppressed to less than 20% HIV-1 positive control with all treatment concentrations and at all infection days (Figure 4).Figure 4


Opposing regulation of endolysosomal pathways by long-acting nanoformulated antiretroviral therapy and HIV-1 in human macrophages.

Araínga M, Guo D, Wiederin J, Ciborowski P, McMillan J, Gendelman HE - Retrovirology (2015)

NanoATV treatment affects HIV-1 reverse transcriptase (RT) activity. HIV-1 RT activity was determined in treated (native ATV or nanoATV) MDM followed by HIV-1 infection at days 0, 5 or 10. HIV-1 infected cells without any treatment served as a positive control for RT activity. All samples were collected after 7 days of viral infection. Results shown are the mean of 5 replicates.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4307176&req=5

Fig4: NanoATV treatment affects HIV-1 reverse transcriptase (RT) activity. HIV-1 RT activity was determined in treated (native ATV or nanoATV) MDM followed by HIV-1 infection at days 0, 5 or 10. HIV-1 infected cells without any treatment served as a positive control for RT activity. All samples were collected after 7 days of viral infection. Results shown are the mean of 5 replicates.
Mentions: To confirm the antiretroviral activity of nanoATV treatment HIV-1 reverse transcriptase (RT) activity was determined in HIV-1-infected human monocyte-derived macrophages (MDM) treated with either native- or nanoATV. Cells were treated with 10, 100 or 250 μM of native- or nanoATV for 16 hours. At this time the medium was removed, cells were washed 3 times with phosphate buffered saline and fresh medium without drug was added prior to HIV-1ADA challenge at a multiplicity of infection (MOI) of 0.1 at days 0, 5 and 10 after treatment. Infected cells were cultured for an additional 7 days and RT activity in the culture medium was determined. Significant differences were found between cells treated with native- or nanoATV. For native ATV treated cells, RT activity was suppressed only in the day 0 infection group, at all treatment concentrations. At 10 and 100 μM little antiviral suppression was observed in the days 5 and 10 infection groups. In contrast, for cells treated with nanoATV, RT activity was suppressed to less than 20% HIV-1 positive control with all treatment concentrations and at all infection days (Figure 4).Figure 4

Bottom Line: NanoART and native ATV treated uninfected cells showed limited effects.The data was confirmed by Western blot.DAVID and KEGG bioinformatics analyses of proteomic data showed relationships between secretory, mobility and phagocytic cell functions and virus and particle trafficking.

View Article: PubMed Central - PubMed

Affiliation: Departments of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, 985880 Nebraska Medical Center, Omaha 68198-5880, NE, USA. hegendel@unmc.edu.

ABSTRACT

Background: Long-acting nanoformulated antiretroviral therapy (nanoART) is designed to improve patient regimen adherence, reduce systemic drug toxicities, and facilitate clearance of human immunodeficiency virus type one (HIV-1) infection. While nanoART establishes drug depots within recycling and late monocyte-macrophage endosomes, whether or not this provides a strategic advantage towards viral elimination has not been elucidated.

Results: We applied quantitative SWATH-MS proteomics and cell profiling to nanoparticle atazanavir (nanoATV)-treated and HIV-1 infected human monocyte-derived macrophages (MDM). Native ATV and uninfected cells served as controls. Both HIV-1 and nanoATV engaged endolysosomal trafficking for assembly and depot formation, respectively. Notably, the pathways were deregulated in opposing manners by the virus and the nanoATV, likely by viral clearance. Paired-sample z-scores, of the proteomic data sets, showed up- and down- regulation of Rab-linked endolysosomal proteins. NanoART and native ATV treated uninfected cells showed limited effects. The data was confirmed by Western blot. DAVID and KEGG bioinformatics analyses of proteomic data showed relationships between secretory, mobility and phagocytic cell functions and virus and particle trafficking.

Conclusions: We posit that modulation of endolysosomal pathways by antiretroviral nanoparticles provides a strategic path to combat HIV infection.

Show MeSH
Related in: MedlinePlus