Limits...
Pulsed stable isotope labeling of amino acids in cell culture uncovers the dynamic interactions between HIV-1 and the monocyte-derived macrophage.

Kraft-Terry SD, Engebretsen IL, Bastola DK, Fox HS, Ciborowski P, Gendelman HE - J. Proteome Res. (2011)

Bottom Line: Synthesis rates of cellular metabolic, regulatory, and DNA packaging activities were decreased, whereas, those affecting antigen presentation (major histocompatibility complex I and II) and interferon-induced antiviral activities were increased.Interestingly, enrichment of proteins linked to chromatin assembly or disassembly, DNA packaging, and nucleosome assembly were identified that paralleled virus-induced cytopathology and replication.We conclude that HIV-1 regulates a range of host MDM proteins that affect its survival and abilities to contain infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, Nebraska 68198-5880, USA.

ABSTRACT
Dynamic interactions between human immunodeficiency virus-1 (HIV-1) and the macrophage govern the tempo of viral dissemination and replication in its human host. HIV-1 affects macrophage phenotype, and the macrophage, in turn, can modulate the viral life cycle. While these processes are linked to host-cell function and survival, the precise intracellular pathways involved are incompletely understood. To elucidate such dynamic virus-cell events, we employed pulsed stable isotope labeling of amino acids in cell culture. Alterations in de novo protein synthesis of HIV-1 infected human monocyte-derived macrophages (MDM) were examined after 3, 5, and 7 days of viral infection. Synthesis rates of cellular metabolic, regulatory, and DNA packaging activities were decreased, whereas, those affecting antigen presentation (major histocompatibility complex I and II) and interferon-induced antiviral activities were increased. Interestingly, enrichment of proteins linked to chromatin assembly or disassembly, DNA packaging, and nucleosome assembly were identified that paralleled virus-induced cytopathology and replication. We conclude that HIV-1 regulates a range of host MDM proteins that affect its survival and abilities to contain infection.

Show MeSH

Related in: MedlinePlus

MDM immunostained for HIV-1p24 protein expression calculated as percent of HIV-1p24 positive cells, expressed as ±1 standard deviation of the mean. Illustrated are (A) control, uninfected MDM and HIV-1-infected MDM on (B) day 3, (C) day 5, and (D) day 7 after viral infection.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3108467&req=5

fig2: MDM immunostained for HIV-1p24 protein expression calculated as percent of HIV-1p24 positive cells, expressed as ±1 standard deviation of the mean. Illustrated are (A) control, uninfected MDM and HIV-1-infected MDM on (B) day 3, (C) day 5, and (D) day 7 after viral infection.

Mentions: To link viral replication to protein identifications, we stained infected cells for HIV-1p24 to document the course of HIV-1ADA infection at the time points examined by proteomics. The percentages of infected cells increased over time and are illustrated in Figure 2. On days 3, 5 and 7, 17.6 ± 3.7, 44.0 ± 9.1 and 70.7 ± 4.4% of the MDM were HIV-1p24 positive, respectively. HIV-1ADA MDM infection levels correlated with the formation of multinucleated giant cells (Figure 2); a stage of infection when cell death is not prominent.(18) It is understood that cell death can result after long-standing infection (2 weeks or longer). This occurs, in part, due to laboratory adaptation of the HIV-1ADA strain used in study. Virus-induced macrophage death is not typical of either primary HIV-1 isolates or with the this current strain when MDM are infected for one week at an MOI of 0.1.


Pulsed stable isotope labeling of amino acids in cell culture uncovers the dynamic interactions between HIV-1 and the monocyte-derived macrophage.

Kraft-Terry SD, Engebretsen IL, Bastola DK, Fox HS, Ciborowski P, Gendelman HE - J. Proteome Res. (2011)

MDM immunostained for HIV-1p24 protein expression calculated as percent of HIV-1p24 positive cells, expressed as ±1 standard deviation of the mean. Illustrated are (A) control, uninfected MDM and HIV-1-infected MDM on (B) day 3, (C) day 5, and (D) day 7 after viral infection.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3108467&req=5

fig2: MDM immunostained for HIV-1p24 protein expression calculated as percent of HIV-1p24 positive cells, expressed as ±1 standard deviation of the mean. Illustrated are (A) control, uninfected MDM and HIV-1-infected MDM on (B) day 3, (C) day 5, and (D) day 7 after viral infection.
Mentions: To link viral replication to protein identifications, we stained infected cells for HIV-1p24 to document the course of HIV-1ADA infection at the time points examined by proteomics. The percentages of infected cells increased over time and are illustrated in Figure 2. On days 3, 5 and 7, 17.6 ± 3.7, 44.0 ± 9.1 and 70.7 ± 4.4% of the MDM were HIV-1p24 positive, respectively. HIV-1ADA MDM infection levels correlated with the formation of multinucleated giant cells (Figure 2); a stage of infection when cell death is not prominent.(18) It is understood that cell death can result after long-standing infection (2 weeks or longer). This occurs, in part, due to laboratory adaptation of the HIV-1ADA strain used in study. Virus-induced macrophage death is not typical of either primary HIV-1 isolates or with the this current strain when MDM are infected for one week at an MOI of 0.1.

Bottom Line: Synthesis rates of cellular metabolic, regulatory, and DNA packaging activities were decreased, whereas, those affecting antigen presentation (major histocompatibility complex I and II) and interferon-induced antiviral activities were increased.Interestingly, enrichment of proteins linked to chromatin assembly or disassembly, DNA packaging, and nucleosome assembly were identified that paralleled virus-induced cytopathology and replication.We conclude that HIV-1 regulates a range of host MDM proteins that affect its survival and abilities to contain infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, Nebraska 68198-5880, USA.

ABSTRACT
Dynamic interactions between human immunodeficiency virus-1 (HIV-1) and the macrophage govern the tempo of viral dissemination and replication in its human host. HIV-1 affects macrophage phenotype, and the macrophage, in turn, can modulate the viral life cycle. While these processes are linked to host-cell function and survival, the precise intracellular pathways involved are incompletely understood. To elucidate such dynamic virus-cell events, we employed pulsed stable isotope labeling of amino acids in cell culture. Alterations in de novo protein synthesis of HIV-1 infected human monocyte-derived macrophages (MDM) were examined after 3, 5, and 7 days of viral infection. Synthesis rates of cellular metabolic, regulatory, and DNA packaging activities were decreased, whereas, those affecting antigen presentation (major histocompatibility complex I and II) and interferon-induced antiviral activities were increased. Interestingly, enrichment of proteins linked to chromatin assembly or disassembly, DNA packaging, and nucleosome assembly were identified that paralleled virus-induced cytopathology and replication. We conclude that HIV-1 regulates a range of host MDM proteins that affect its survival and abilities to contain infection.

Show MeSH
Related in: MedlinePlus