Human immunodeficiency virus type 1 gp120 and other activation stimuli are highly effective in triggering alpha interferon and CC chemokine production in circulating plasmacytoid but not myeloid dendritic cells - PubMed (original) (raw)

Human immunodeficiency virus type 1 gp120 and other activation stimuli are highly effective in triggering alpha interferon and CC chemokine production in circulating plasmacytoid but not myeloid dendritic cells

Manuela Del Cornò et al. J Virol. 2005 Oct.

Abstract

Exposure to aldrithiol-2-inactivated human immunodeficiency virus type 1 or gp120, but not gp41, triggered alpha interferon (IFN-alpha), CC chemokine ligand 2 (CCL2), CCL3, and CCL4 production in human plasmacytoid dendritic cells (DCs) but not in myeloid DCs (M-DCs) or monocyte-derived DCs from the same donors. The nonresponsiveness of M-DCs for IFN-alpha/beta production was a general feature specific to these cells, as they also failed to produce it in response to inactivated influenza virus, poly(I-C), lipopolysaccharide, Staphylococcus aureus Cowans I, or CD40L. The different capacities of circulating DC subsets to produce immune mediators in response to most stimuli argue for a different role for these cells in the regulation of innate immunity to pathogens.

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Figures

FIG. 1.

FIG. 1.

IFN-α/β production by circulating DC subsets and in vitro-generated myeloid DCs exposed to AT-2-inactivated X4 and R5 HIV-1 strains. (A) P-DCs (2.5 × 105/ml), M-DCs (2.5 × 105/ml), and MD-DCs (1 × 106/ml) were cultured in a 96-well, flat-bottomed tissue culture plate in 5 to 10% fetal bovine serum-RPMI 1640 medium and exposed to X4 (HIV-1IIIB) and R5 (HIV-1BaL) AT-2-inactivated HIV strains (dose equivalent to a multiplicity of infection of 0.1 before inactivation). Twenty-four hours later, culture supernatants were collected and the IFN-α content was measured by enzyme-linked immunosorbent assay (ELISA) (Pierce Endogen, Rockford, IL; detection limit, 12.5 pg/ml). A representative experiment out of six performed is shown. (B) The content of biologically active IFN-α/β was determined in the supernatants collected from the same cultures described above by a vesicular stomatitis virus cytopathic reduction assay. A representative experiment out of three performed is shown.

FIG. 2.

FIG. 2.

Effect of recombinant gp120 and gp41 on the production of IFN-α by P-DCs, M-DCs, and MD-DCs. DC subsets were treated with 2 μg/ml of recombinant X4 or R5 HIV-1 gp120 (HIV-1IIIB and HIV-1BaL, obtained from the EU program EVA/MRC and the National Institutes of Health AIDS Research and Reference Reagent Program, respectively) or X4 gp41 (HIV-1IIIB, purchased from ABI, Columbia, MD) for 24 h. Cell supernatants were collected, and the IFN-α content was measured by ELISA. A representative experiment out of three performed is shown.

FIG. 3.

FIG. 3.

IFN-α/β production by P-DCs, M-DCs, and MD-DCs in response to different IFN-inducing stimuli. (A) DC subsets were stimulated with inactivated influenza virus (strain A/Moscow/10/99, 20 ng/ml of hemagglutinin), poly(I-C) (pI:C; 20 μg/ml; Sigma-Aldrich, St. Louis, MO), LPS (100 ng/ml; Sigma-Aldrich), SAC (1:5,000; Calbiochem, San Diego, CA), and CD40L-transfected J558 cells at a ratio of 5:1 (23) or left untreated. Cell supernatants were collected 24 h later, and the content of IFN-α was determined by ELISA. The mean values ± standard deviations of six independent experiments are shown. (B) The content of biologically active IFN-α/β was determined in the supernatants collected from the same cultures as described in the legend to Fig. 1. A representative experiment out of three performed is shown.

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