M1 polarization of human monocyte-derived macrophages restricts pre and postintegration steps of HIV-1 replication

Luca Cassetta, Anna Kajaste-Rudnitski, Tiziana Coradin, Elisa Saba, Giulia Della Chiara, Marialuisa Barbagallo, Francesca Graziano, Massimo Alfano, Edana Cassol, Elisa Vicenzi, Guido Poli

Research output: Contribution to journalArticle

Abstract

Objective: Functional polarization of human monocyte-derived macrophages (MDMs) into M1 cells leads to inhibition of R5 HIV-1 replication and viral DNA synthesis in comparison to control, unpolarized cells together with CD4 downregulation from the cell surface and upregulation of CCR5-binding chemokine secretion. We here investigated whether a postentry restriction of virus replication is also induced by M1 polarization of MDM. Design: MDMwere first polarized to M1 cells by 18 h stimulation with interferon-γ and tumor necrosis factor-α; the cytokines were then removed and the cells were infected with vesicular stomatitis virus G-protein pseudotyped enhanced green fluorescence protein HIV-1 (HIV-GFP) generating a single-round infection cycle. Methods: HIV-1 expression was monitored in terms of eGFP expression by fluorescence activated cell sorter (FACS) analysis and real-time PCR analysis of total HIV-1 gag DNA, 2-long terminal repeat DNA, proviral DNA, and multiply spliced RNA transcripts. Expression of apolipopoprotein B mRNA-editing, enzyme-catalytic, polypeptide-like 3G (APOBEC3G), and APOBEC3A was tested by western blotting and FACS analysis. Results: Inhibition of HIV-GFP expression was observed in M1-MDM along with impaired viral DNA synthesis, delayed proviral integration, and reduced proviral transcription. Although APOBEC3G levels were similar in M1 and unpolarized MDM, APOBEC 3A was selectively expressed only by M1 cells. Conclusion: M1 polarization of in-vitro differentiated primary MDM determines a transient, but profound restriction of HIV-1 replication affecting multiple (entry and postentry) steps in the virus life cycle likely involving the upregulated expression of APOBEC3A.

Original languageEnglish
Pages (from-to)1847-1856
Number of pages10
JournalAIDS (London, England)
Volume27
Issue number12
DOIs
Publication statusPublished - Jul 31 2013

Fingerprint

HIV-1
Macrophages
Fluorescence
Viral DNA
DNA
HIV
Messenger RNA
Peptides
Terminal Repeat Sequences
Enzymes
Virus Replication
Life Cycle Stages
Chemokines
Interferons
Real-Time Polymerase Chain Reaction
Up-Regulation
Down-Regulation
Tumor Necrosis Factor-alpha
Western Blotting
RNA

Keywords

  • APOBEC3A
  • APOBEC3G
  • Cytokines
  • HIV-1
  • Integration
  • M1 polarization
  • Macrophage
  • Transcription

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Infectious Diseases

Cite this

M1 polarization of human monocyte-derived macrophages restricts pre and postintegration steps of HIV-1 replication. / Cassetta, Luca; Kajaste-Rudnitski, Anna; Coradin, Tiziana; Saba, Elisa; Della Chiara, Giulia; Barbagallo, Marialuisa; Graziano, Francesca; Alfano, Massimo; Cassol, Edana; Vicenzi, Elisa; Poli, Guido.

In: AIDS (London, England), Vol. 27, No. 12, 31.07.2013, p. 1847-1856.

Research output: Contribution to journalArticle

Cassetta, Luca ; Kajaste-Rudnitski, Anna ; Coradin, Tiziana ; Saba, Elisa ; Della Chiara, Giulia ; Barbagallo, Marialuisa ; Graziano, Francesca ; Alfano, Massimo ; Cassol, Edana ; Vicenzi, Elisa ; Poli, Guido. / M1 polarization of human monocyte-derived macrophages restricts pre and postintegration steps of HIV-1 replication. In: AIDS (London, England). 2013 ; Vol. 27, No. 12. pp. 1847-1856.
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AU - Saba, Elisa

AU - Della Chiara, Giulia

AU - Barbagallo, Marialuisa

AU - Graziano, Francesca

AU - Alfano, Massimo

AU - Cassol, Edana

AU - Vicenzi, Elisa

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