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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.

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(A) IFN signaling pathway coded with identified proteins on day 7 postinfection. Gray proteins were identified but did not reach statistical significance. (B) Ingenuity Pathway Analysis highest scoring interaction network. Increased synthesis rates are indicated by increasing intensity of red; decreased synthesis rates are denoted in green.
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fig4: (A) IFN signaling pathway coded with identified proteins on day 7 postinfection. Gray proteins were identified but did not reach statistical significance. (B) Ingenuity Pathway Analysis highest scoring interaction network. Increased synthesis rates are indicated by increasing intensity of red; decreased synthesis rates are denoted in green.

Mentions: A complete list of all significant proteins and their corresponding fold change are included in Supplemental Table 1 (Supporting Information). The highest scoring interaction network identified by IPA for day 7 of infection identified significant upregulation in de novo synthesis of IFN-induced proteins (Figure 4), therefore, indicating that the rate of antiviral response by the macrophage increased as HIV-1 infection progressed. These antiviral proteins affect different points during the viral life cycle (Supplemental Figure 1, Supporting Information). Although numerous antiviral proteins were up-regulated [including: 2′-5′-oligadenylate sythetase 2 (2–5 OAS2), signal transducers and activators of transcription 1 (STAT1), IFN-induced guanylate-binding protein 1 and 2 (GBP1, GBP2), IFN-induced GTP-binding protein Mx1 (Mx1), and bone marrow stromal antigen 2 (BST2 or tetherin)], virus production increased over time, therefore allowing cells to secrete progeny virus or to spread infection to nearby cells (Supplemental Figure 1). While it is known that MCSF-differentiated macrophages display an M2 anti-inflammatory phenotype,(34) it is known that HIV-1 induces an M1 inflammatory phenotype at early stages of infection.(6) This commonly results in IFN production and secretion of pro-inflammatory factors5,35 demonstrating innate antiretroviral responses of MDM.


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)

(A) IFN signaling pathway coded with identified proteins on day 7 postinfection. Gray proteins were identified but did not reach statistical significance. (B) Ingenuity Pathway Analysis highest scoring interaction network. Increased synthesis rates are indicated by increasing intensity of red; decreased synthesis rates are denoted in green.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: (A) IFN signaling pathway coded with identified proteins on day 7 postinfection. Gray proteins were identified but did not reach statistical significance. (B) Ingenuity Pathway Analysis highest scoring interaction network. Increased synthesis rates are indicated by increasing intensity of red; decreased synthesis rates are denoted in green.
Mentions: A complete list of all significant proteins and their corresponding fold change are included in Supplemental Table 1 (Supporting Information). The highest scoring interaction network identified by IPA for day 7 of infection identified significant upregulation in de novo synthesis of IFN-induced proteins (Figure 4), therefore, indicating that the rate of antiviral response by the macrophage increased as HIV-1 infection progressed. These antiviral proteins affect different points during the viral life cycle (Supplemental Figure 1, Supporting Information). Although numerous antiviral proteins were up-regulated [including: 2′-5′-oligadenylate sythetase 2 (2–5 OAS2), signal transducers and activators of transcription 1 (STAT1), IFN-induced guanylate-binding protein 1 and 2 (GBP1, GBP2), IFN-induced GTP-binding protein Mx1 (Mx1), and bone marrow stromal antigen 2 (BST2 or tetherin)], virus production increased over time, therefore allowing cells to secrete progeny virus or to spread infection to nearby cells (Supplemental Figure 1). While it is known that MCSF-differentiated macrophages display an M2 anti-inflammatory phenotype,(34) it is known that HIV-1 induces an M1 inflammatory phenotype at early stages of infection.(6) This commonly results in IFN production and secretion of pro-inflammatory factors5,35 demonstrating innate antiretroviral responses of MDM.

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