1. Cell Biology
  2. Microbiology and Infectious Disease
Download icon

Tuberculosis-associated IFN-I induces Siglec-1 on tunneling nanotubes and favors HIV-1 spread in macrophages

  1. Maeva Dupont
  2. Shanti Souriant
  3. Luciana Balboa
  4. Thien-Phong Vu Manh
  5. Karine Pingris
  6. Stella Rousset
  7. Céline Cougoule
  8. Yoann Rombouts
  9. Renaud Poincloux
  10. Myriam Ben Neji
  11. Carolina Allers
  12. Deepak Kaushal
  13. Marcelo J Kuroda
  14. Susana Benet
  15. Javier Martinez-Picado
  16. Nuria Izquierdo-Useros
  17. Maria del Carmen Sasiain
  18. Isabelle Maridonneau-Parini
  19. Olivier Neyrolles
  20. Christel Vérollet  Is a corresponding author
  21. Geanncarlo Lugo-Villarino  Is a corresponding author
  1. Institut de Pharmacologie et Biologie Structurale, IPBS, Université de Toulouse, CNRS, UPS, France
  2. International associated laboratory (LIA) CNRS 'IM-TB/HIV', France
  3. Institute of Experimental Medicine-CONICET, National Academy of Medicine, Argentina
  4. Aix Marseille Univ, CNRS, INSERM, CIML, France
  5. Tulane National Primate Research Center, Department of Microbiology and Immunology, School of Medicine, Tulane University, United States
  6. IrsiCaixa AIDS Research Institute, Department of Retrovirology, Spain
  7. Universitat Autònoma de Barcelona, Spain
  8. University of Vic-Central University of Catalonia (UVic-UCC), Spain
  9. Catalan Institution for Research and Advanced Studies (ICREA), Spain
  10. Institut d’Investigació en Ciències de la Salut Germans Trias i Pujol, Spain
Research Article
Cite this article as: eLife 2020;9:e52535 doi: 10.7554/eLife.52535
Voice your concerns about research culture and research communication: Have your say in our 7th annual survey.
7 figures, 1 table and 2 additional files

Figures

Figure 1 with 2 supplements
Tuberculosis-associated microenvironments induce Siglec-1 expression in macrophages.

(A–D) For 3 days, human monocytes were differentiated into macrophages with cmCTR (white) or cmMTB (black) supernatants. (A) Heatmap from a transcriptomic analysis (GEO submission GSE139511) illustrating the top 60 differentially expressed genes (DEGs) between cmCTR- or cmMTB-cells. Selection of the top DEGs was performed using an adjusted p-value ≤ 0.05, a fold change of at least 2, and a minimal expression of 6 in a log2 scale. Hierarchical clustering was performed using the complete linkage method and the Pearson correlation metric with Morpheus (Broad Institute). Interferon-stimulated genes (ISG) are labelled with an asterisk and Siglec-1 is indicated in red. (B–D) Validation of Siglec-1 expression in cmMTB-treated macrophages. Vertical scatter plots showing the relative abundance to mRNA (B), median fluorescent intensity (MFI) (C), and mean number of Siglec-1 antibody binding sites per cell (D). Each circle represents a single donor and histograms median values. (E) Representative immunohistochemical images of Siglec-1 staining (brown) in lung biopsies of healthy, SIV infected (SIV), active TB (ATB), and co-infected (ATB-SIV) non-human primates (NHP). Scale bar, 100 µm. Insets are 2x zoom. (F) Vertical Box and Whisker plot indicating the distribution of the percentage of Siglec-1+ alveolar macrophages in lung biopsies from the indicated NHP groups. Quantification analysis from n = 800 alveolar macrophages grouped from three independent animals per NHP group. (G) Vertical scatter plots displaying the number of cells that are positive for Siglec-1 per mm² of lung biopsies from the indicated NHP groups. Each symbol represents a single animal per NHP group. (H) Correlation between Siglec-1+ cells per mm² of lung tissue and the pathological score for healthy (white circle), SIV+ (white triangles), latent (black circle) or active (black square) TB, and SIV+ with latent (grey circle) or active (grey square) TB. Each symbol represents a single animal per NHP group. Mean value is represented as a black line. Statistical analyses: Two-tailed, Wilcoxon signed-rank test (B–D), Mann-Whitney unpaired test (F–G), Spearman correlation (H). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ns: not significant. See Figure 1—source data 1.

Figure 1—source data 1

Raw data and statistical analyses supporting Siglec-1 expression in human and non-human primate macrophages exposed to TB-associated microenvironment.

https://cdn.elifesciences.org/articles/52535/elife-52535-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Tuberculosis-associated microenvironments increase Siglec-1 expression in human macrophages.

(A–D) For 3 days, human monocytes were differentiated into macrophages with cmCTR (white) or cmMTB (black) supernatants. (A) (Left) Gene set enrichment plot of the interferon alpha (IFNα) response (hallmark collection of MSigDB). This plot shows the distribution of the barcode between macrophages exposed to cmCTR (red) versus cmMTB (blue) supernatants. Each bar of the barcode corresponds to a signature gene of the gene set. The skewing to the right indicates enrichment in macrophages exposed to cmMTB versus cmCTR supernatant of genes up-regulated in response to IFNα. (Right) Gene set enrichment plot of the IFNγ response (hallmark collection of MSigDB). (B) Flow cytometry gating strategy to assess Siglec-1 cell-surface expression in human macrophages exposed to cmCTR (white) and cmMTB (black). (Left) Based on size (FSC-A) and granularity (SSC-A), a gate was created to separate human macrophages from cell debris and dying cells. Macrophages were then subjected through a second gate based FSC Area and Height Scaling (FSC-A and FSC-H) to separate singlets from doublets. (Right) Based on the singlet gate, the histogram plot illustrates Siglec-1 expression that is higher in cmMTB- (black) than in cmCTR-treated (white) macrophages. (C) Representative immunofluorescence of Siglec-1 intracellular staining (green), actin (red) and nuclei (blue) after 3 days of monocyte conditioning with cmMTB. Scale bar, 10 μm. (D) Vertical scatter plot showing the quantification of Siglec-1 intracellular staining in cmCTR- or cmMTB-treated cells in the presence of an IFNAR-2 blocking (α-IFNAR) or control (α-IgG) antibodies during cell conditioning. Each circle represents a single donor and histograms median value. (E) Median fluorescence intensity (MFI) of Siglec-1 cell-surface expression in human macrophages exposed to cmCTR and infected with HIV-1 assessed by flow cytometry. The histogram plot illustrates Siglec-1 expression that increases within days post-HIV-1 infection of cmCTR-macrophages. Statistical analyses: Two-tailed, Wilcoxon signed-rank test (D). *p<0.05, **p<0.01, ns: not significant.

Figure 1—figure supplement 2
Tuberculosis-associated microenvironments increase Siglec-1 expression in non-human primate alveolar macrophages.

(A) Accumulation of Siglec-1+ alveolar macrophages in the lung of co-infected non-human primates (NHP). Representative immunohistochemical images of Siglec-1 staining (brown) in lung biopsies of healthy, SIV-infected (SIV), active tuberculosis (ATB) and co-infected with SIV (ATB-SIV) NHP. Scale bars from top to bottom: 2 mm, 500 µm and 50 µm. (B–C) Siglec-1+ cells display the alveolar macrophage morphology. (B) Representative immunohistochemistry image from lung biopsy of an ATB-SIV NHP stained for Siglec-1 (brown). Siglec-1+ cells display a cell morphology with a single nucleus and large cytoplasm reminiscent of macrophage (black arrowhead); Siglec-1- cells display a different nucleus morphology and small cytoplasm reminiscent of neutrophils (red arrowhead). Scale bar, 20 µm. (C) Representative immunofluorescence images of alveolar macrophages found in lung biopsy of a representative ATB-SIV NHP stained for Siglec-1 (red), CD163 (green) and nuclei (DAPI, blue). Scale bar, 20 µm.

Siglec-1 expression is dependent on Mtb-induced type I IFN signaling.

(A) Vertical scatter plot showing the relative abundance of IFN-I in cmCTR (white) and cmMTB (black) media, as measured indirectly after 24 hr exposure to the HEK-Blue IFN-α/β reporter cell line yielding reporter activity in units (U) per mL. (B–D) Vertical scatter plots displaying the median fluorescent intensity (MFI) of Siglec-1 cell-surface expression after three days of monocyte differentiation into macrophages either with cmMTB (black) or cmCTR (white), the indicated recombinant cytokines (B), the presence of an IL-10 depletion (α-IL-10) or a control (α-IgG) antibodies (C), or the presence of an IFNAR-2 blocking (α-IFNAR) or control (α-IgG) antibodies (D). (E) Representative serial immunohistochemical images of lung biopsies of a co-infected (ATB-SIV) NHP stained for Siglec-1 (brown, top) and pSTAT1 (brown, bottom). Scale bar, 250 µm. Insets are 10x zooms. (F) Correlation of the percentage of cells positive for Siglec-1 and pSTAT1, as measured per mm2 of lung tissue from the indicated NHP groups. Mean value is represented as a black line. (G) Representative immunohistochemical images of lung biopsies from the indicated NHP group stained for pSTAT1 (brown). Arrowheads show pSTAT1-positive nuclei. Scale bar, 500 µm. (H) Upper panel: Vertical Box and Whisker plot illustrating the percentage of pSTAT1+ alveolar macrophages in lung biopsies from the indicated NHP groups. Quantification analysis from n = 600 alveolar macrophages grouped from three independent animals per NHP group. Lower panel: Correlation of the percentage of alveolar macrophages positive for Siglec-1 and pSTAT1, from the indicated NHP groups. Mean value is represented as a black line. (A–D) Each circle within vertical scatter plots represents a single donor and histograms median value. Statistical analyses: Two-tailed, Wilcoxon signed-rank test (A–D), Spearman correlation (F, H lower panel), and Mann-Whitney unpaired test (H, upper panel). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ns: not significant. See Figure 2—source data 1.

Figure 2—source data 1

Raw data and statistical analyses supporting that IFN-I induced by M. tuberculosis is responsible for Siglec-1 expression in human and non-human primate macrophages.

https://cdn.elifesciences.org/articles/52535/elife-52535-fig2-data1-v2.xlsx
Figure 3 with 3 supplements
Siglec-1 localization on thick TNT is associated with their length and HIV-1/mitochondria cargo.

(A–F) Human monocytes were differentiated into macrophages with cmMTB for 3 days, and then infected with HIV-1-ADA strain (unless indicated otherwise) and fixed 3 days post-infection. (A) Representative immunofluorescence images of cmMTB-treated macrophages infected with HIV-1-ADA, and stained for extracellular Siglec-1 (red), intracellular tubulin (MT, green) and Wheat Germ Agglutinin (WGA, blue). Inserts are 3x zooms. Red arrowheads show Siglec-1 localization on TNT. Scale bar, 20 µm. (B) Vertical bar plot showing the semi-automatic quantification of Siglec-1+ TNT (black) and Siglec-1- TNT (white) in thick (WGA+, MT+) and thin (WGA+, MT-) TNT. 400 TNT were analyzed from two independent donors. (C) Siglec-1+ TNT exhibit a larger length index. Violin plots displaying the semi-automatic quantification of TNT length (in μm) for thin (WGA+, MT-), and thick TNT (WGA+, MT+) expressing Siglec-1 or not. 400 TNT were analyzed per condition from two independent donors. (D) Representative immunofluorescence images of cmMTB-treated macrophages 3 day post-infection with HIV-1-NLAD8-VSVG strain, and stained for extracellular Siglec-1 (red), intracellular HIV-1Gag (green) and WGA (grey). Scale bar, 10 µm. (E) Vertical bar plots indicating the quantification of presence (dark grey) or absence (light grey) of HIV-1Gag in thick TNT (WGA+, MT+) expressing Siglec-1 or not. 120 TNT in at least 1000 cells were analyzed from four independent donors. (F) Representative immunofluorescence images of cmMTB-treated macrophages infected with HIV-1-ADA loaded with MitoTracker (MitoT, green), and stained for extracellular Siglec-1 (red) and WGA (grey). Inserts are 3x zooms. Green arrowheads show mitochondria inside TNT. Scale bar, 10 µm. Statistical analyses: Two-way ANOVA comparing the presence of Siglec-1 in thin and thick TNT (B), and two-tailed Mann-Whitney unpaired test comparing TNT length (C) and the presence of HIV-1 in TNT (E). *p<0.05, ****p<0.0001. See Figure 3—source data 1.

Figure 3—source data 1

Raw data and statistical analyses supporting Siglec-1 expression on thick TNT and its correlation with TNT length.

https://cdn.elifesciences.org/articles/52535/elife-52535-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Siglec-1 localizes specifically on thick tunneling nanotubes that contain HIV-1Gag and mitochondria.

(A–C) Human monocytes were differentiated into macrophages with cmMTB for 3 days, infected with HIV-1-ADA strain (unless indicated otherwise) and then fixed at day 3 (A–B) or 14 (C) post-infection. (A) Representative immunofluorescence images used for semi-automatic quantification of TNT in cmMTB-treated macrophages infected with HIV-1. Cells were stained for extracellular Siglec-1 (red), intracellular tubulin (MT, grey) and Wheat Germ Agglutinin (WGA, not shown). Blue lines show all TNT considered. Thick (WGA+, MT+) and thin (WGA+, MT-) TNT were assessed for Siglec-1 positivity by applying a threshold and measured in length. Scale bar, 20 µm. (B) Representative immunofluorescence images of cmMTB-treated macrophages infected with HIV-1-NLAD8-VSVG, loaded with Mitotracker (MitoT, red) and stained for intracellular HIV-1Gag (green) and WGA (grey). Red arrowheads show mitochondria inside HIV-1Gag-containing TNT. Inserts are 2x zoom. Scale bar, 20 µm. (C) Representative immunofluorescence images of cmMTB-treated macrophages infected with HIV-1 and kept in culture until day 14. Cells were fixed and stained for intracellular HIV-1Gag (green), extracellular Siglec-1 (red) and WGA (grey). Red arrowheads show Siglec-1 on HIV-1Gag-containing TNT emanating from an infected multinucleated giant cell (MGC). Scale bar, 20 µm.

Figure 3—video 1
Related to Figure 3A.

Z-stack of confocal microscopy images, showing Siglec-1 (red), microtubules (MT, green) and F-actin (grey) of day 6 HIV-1-infected macrophages, treated with cmMTB. Siglec-1 localizes on the thick MT+ F-actin+ TNT but not on thin MT- F-actin+ TNT.

Figure 3—video 2
Related to Figure 3D.

3D reconstitution of confocal images, showing Siglec-1 (red), HIV-1Gag (green) and WGA (grey) of day 6 HIV-1-infected macrophages, treated with cmMTB.

Figure 4 with 1 supplement
The exacerbation of HIV-1 infection and spread in macrophages treated with cmMTB requires Siglec-1.

(A) Experimental design. Monocytes from healthy subjects were transfected with siRNA targeting of Siglec-1 (siSiglec-1, black) or not (siCtrl, white). A day after, monocytes were differentiated into macrophages with cmMTB for 3 days. Cells were then infected with HIV-1-ADA (blue protocol) to measure the formation (B) and length (C) of TNT at day 7, or assess HIV-1 production and multinucleated giant cell (MGC) formation at day 14 (F–G). In parallel, cells were either infected with HIV-NLAD8-VSVG or labelled with mitoTracker to measure the transfer (red protocol) of HIV-1 (D) or mitochondria (E), respectively. (B) Before-and-after plots showing the percentage of cells forming thick TNT (F-actin+, WGA+, MT+). (C) Violin plots displaying the semi-automatic quantification of TNT length (in μm) for thick (WGA+, MT+) TNT; 300 TNT were analyzed per condition from two independent donors. (D–E) Before-and-after plots indicating the percentage of HIV-1Gag+ cells (D) or MitoTracker+ cells (E) among CellTracker+ cells after 24 hr co-culture. (F) Representative immunofluorescence images of siRNA transfected cells treated with cmMTB, 14 days post-HIV-1 infection. Cells were stained for intracellular HIV-1Gag (green), F-actin (red) and DAPI (blue). Scale bar, 500 µm. (G) Vertical scatter plots showing HIV-1-p24 concentration in cell supernatants (upper panel) and percentage of MGC (lower panel) at day 14 post-HIV-1 infection in cells represented in F (siSiglec-1, black; siCtrl, white). (B, D, E and G) Each circle represents a single donor and histograms median value. Statistical analyses: Paired t-test (B, G lower panel) or two-tailed, Wilcoxon signed-rank test (C-E, G upper panel). *p<0.05, **p<0.01, ****p<0.0001. See Figure 4—source data 1.

Figure 4—source data 1

Raw data and statistical analyses supporting a role for Siglec-1 in TNT length, HIV-1 and mitochondrial cell-to-cell trasfer, and exacerbation of HIV-1 infection.

https://cdn.elifesciences.org/articles/52535/elife-52535-fig4-data1-v2.xlsx
Figure 4—figure supplement 1
Siglec-1 is required for the capture and transfer of HIV-1 in cmMTB-treated macrophages.

(A–D, F–G) Monocytes from healthy subjects were transfected with siRNA targeting of Siglec-1 (siSiglec-1, black) or not (siCtrl, white). A day after, monocytes were differentiated into macrophages with cmMTB for 3 days. (A) Representative histogram (left) and vertical scatter plot showing the median fluorescent intensity (MFI) (right) of Siglec-1 expression on the indicated cell populations. (B) Vertical scatter plot indicating the percentage of cells forming TNT. (C–E) Inhibition of Siglec-1 reduces binding of HIV-1-Gag−eGFP virus like particles (GFP-VLP). (C) Representative immunofluorescence images of cmMTB-treated cells incubated with GFP-VLP (green) for 3.5 hr. Cells were fixed and stained for extracellular Siglec-1 (red) and Wheat Germ Agglutinin (WGA, blue). Scale bar, 500 µm. (D) Representative histogram (left) and vertical scatter plot showing the median fluorescent intensity (MFI) (right) displaying of GFP-VLP binding in the indicated cell populations. (E) Vertical scatter plot showing the percentage of GFP-VLP binding in cmMTB treated cells pre-incubated with specific anti-Siglec-1 (α-Siglec-1, grey), anti-Isotype control antibodies (α-IgG, black) or mock (white). (F) Schematics of the experimental procedure for material (HIV-1 and mitochondria) transfer experiments. (G) Vertical scatter plot showing the percentage of HIV-1Gag+ cells at the time of co-culture experiment in the indicated cells. Statistical analyses: Two-tailed, Wilcoxon matched-pairs signed rank test (A, B, D-E, G). *p<0.05, ***p<0.001. ns: not significant.

Figure 4—figure supplement 1—source data 1

Supplemental raw data and statistical analyses supporting the functional role of Siglec-1 in human macrophages using an siRNA-mediated gene silencing approach.

https://cdn.elifesciences.org/articles/52535/elife-52535-fig4-figsupp1-data1-v2.xlsx
Author response image 1
Immunofluorescence analysis of cmMTB-treated cells positive for Siglec-1.

For 3 days, monocytes were differentiated into macrophages either with cmCTR (white) or with cmMTB in the presence of an IFNAR-2 blocking (α-IFNAR, stripped) or control (α-IgG, black) antibodies. Immunofluorescence analyses were performed and the percentage of cells positive for intracellular Siglec-1 were quantified. n=4 donors.

Author response image 2
Correlation of alveolar macrophages that are positive for either Siglec-1 or phospho-STAT1 in NHP (Related to Figure 2H, bottom).

Each symbol represents an image field containing the lung alveoli that was obtained from the three independent animals per NHP group. Mean value is represented as a red line.

Author response image 3
Inactivation of Siglec-1 by siRNA-mediated gene silencing does not affect the expression of M(IL-10) markers in cmMTB-treated cells (Related to Figure 4—figure supplement 1A).

Monocytes from healthy subjects were transfected with siRNA targeting of Siglec-1 (siSiglec-1, black) or not (siCtrl, white). A day after, monocytes were differentiated into macrophages with cmMTB for 3 days. Vertical scatter plots showing the median fluorescent intensity (MFI, upper panels) of Siglec-1, CD16 and CD163, and the percentage of cells expressing Siglec-1, CD16 and CD163 (lower panels). Each circle represents a single donor and histograms median values. n=3 donors.

Tables

Key resources table
Reagent type
(species) or resource
DesignationSource or referenceIdentifiersAdditional information
M. tuberculosisH37RvDerived from E.R. Baldwin's human-lung isolate H37 by W. Steenken New York, United States, 1934(ATCC 25618)
HIV-1ADAGift from Dr. S Benichou
Institut Cochin, Paris, France
N/A
HIV-1NLAD8Gift from Dr. S Benichou
Institut Cochin, Paris, France
N/A
HIV-1ADA Gag-iGFP-VSVGThis paperThis paper
Buffy CoatLeukocytesEtablissement Français du Sang, Toulouse, FranceN/A
Lung biopsies from rhesus macaquesHistological slidesTulane National Primate Research CenterN/A
Cell line (human HeLa JC.53)TZM-blNIH AIDS Reagent ProgramCat# 8129Cultured in: DMEM, 90%; FBS, 10%; 100 units of Penicillin and 0.1 mg/mL of Streptomycin
Cell line (human)HEK-293TNIH AIDS Reagent ProgramCat# 3318Cultured in: DMEM, 10% FCS
Cell line (human)HEK-Blue IFN-α/β CellsInvivogenCat# hkb-ifnabCultured in: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 µg/mL streptomycin, 100 µg/mL Normocin
Transfected construct (human)siRNA to Siglec-1 (SMART-Pool)Horizon DiscoveryCat# L-017521-01-0020(200 nM)
Transfected construct (human)siRNA scramble (SMART Pool)Horizon DiscoveryCat# D-001810-10-50(200 nM)
AntibodyMouse monoclonal anti‑human Siglec-1 (clone 7-293)BiolegendCat# 346008
RRID:AB_11147948
FACS (1 µg/mL)
AntibodyMouse monoclonal anti‑human CD16 (clone 3G8)BiolegendCat# 302019 and 302018; RRID:AB_492974 andAB_314218FACS (1 µg/mL)
AntibodyMouse monoclonal anti‑human CD163 (clone GHI/61)BiolegendCat# 333608
RRID:AB_2228986
FACS (1 µg/mL)
AntibodyMouse monoclonal anti‑human MerTK (clone 590H11G1E3)BiolegendCat# 367607
RRID:AB_2566400
FACS (1 µg/mL)
AntibodyRabbit monoclonal anti‑human STAT1 (clone 42H3)Cell Signaling TechnologyCat# 9175
RRID:AB_2197984
WB (1:100)
AntibodyRabbit anti‑human actin (a.a. 20‑33)Sigma‑AldrichCat# A5060
RRID:AB_476738
WB (1:100)
AntibodyRabbit polyclonal anti-a-tubulinAbcamCat# ab18251
RRID:AB_2210057
IF (5 µg/mL)
AntibodyMouse monoclonal anti-Siglec-1 (clone hsn 7D2)Novus BiologicalsCat# NB 600-534
RRID:AB_526814
IF (10 µg/mL) IHC (1:200)
AntibodyMouse monoclonal anti-Gag RD1 (clone KC57)NIH AIDS Reagent programCat# 13449IF (1:200)
AntibodyMouse monoclonal anti-HIV-1 p24 (clone 183-H12-5C)NIH AIDS Reagent ProgramCat# 3537ELISA (2.5 µg/mL)
AntibodyHuman polyclonal anti-HIV Immune Globulin (HIVIG)NIH AIDS Reagent ProgramCat# 3957ELISA (6.25 µg/mL)
AntibodyPolyclonal goat anti-human IgGSigma-AldrichCat# A0170ELISA (1:10000)
AntibodyMouse monoclonal anti‑human CD163 (clone 10D6)Leica/NovocastraCat# NCL-L-CD163
RRID:AB_2756375
IHC (1:100)
AntibodyAnti-pSTAT1Cell Signaling TechnologyCat# 9167
RRID:AB_561284
WB (1:100)
AntibodyMouse monoclonal anti-IFNAR2 (clone MMHAR-2)Thermo Fisher ScientificCat# 213851
RRID:AB_223508
Blocking (20 µg/mL)
FACS (1 µg/mL)
AntibodyMouse IgG2a isotype controlThermo Fisher ScientificCat# 02-6200
RRID:AB_2532943
Blocking (20 µg/mL)
IF (0.6 µg/mL)
AntibodyPolyclonal F(ab)2 goat anti‑rabbit IgG, AlexaFluor 555Thermo Fisher ScientificCat# A-21430
RRID:AB_2535851
IF (2 µg/mL)
AntibodyPolyclonal F(ab)2 goat anti‑mouse IgG, AlexaFluor 488Thermo Fisher ScientificCat# A-10684
RRID:AB_2534064
IF (2 µg/mL)
AntibodyPlyclonal F(ab)2 goat anti‑mouse IgG, AlexaFluor 555Cell Signaling TechnologyCat# 4409
RRID:AB_1904022
IF (2 µg/mL)
AntibodyPolyclonal goat anti‑rabbit IgG, HRPThermo Fisher ScientificCat# 32460
RRID:AB_1185567
WB (1:10000)
AntibodyPolyclonal goat anti‑mouse IgG, HRPThermo Fisher ScientificCat# 31430
RRID:AB_228307
WB (1:10000)
Cytokine (recombinant, human)M-CSFPeprotechCat# 300‑25(20 ng/mL)
Cytokine (recombinant, human)IFNbPeprotechCat# 300-02BC10 and 100 U/mL
Cytokine (recombinant, human)IL-10PeprotechCat# 200-1010 ng/mL
Monocyte isolationMouse anti‑human CD14 microbeadsMiltenyi BiotecCat# 130‑050‑201
Monocyte isolationLS magnetic columnsMiltenyi BiotecCat# 130‑042‑401
Western blotAmersham ECL Prisme Western Blotting Detection ReagentGE HealthcareCat# RPN2232
Western blotSuperSignal WestPico Chemiluminescent SubstrateThermo ScientificCat# 34080
ELISAIL‑10 ELISA setBD BioscienceCat# 555157
Cell cultureTrypsin EDTA 0.05%Thermo Fisher ScientificCat# 25200072
Cell cultureAccutaseSigma-AldrichCat# A-6964
ProbePhalloidin AlexaFluor 488Thermo Fisher ScientificCat# A12379(33 mM)
ProbePhalloidin Alexa Fluor 647Thermo Fisher ScientificCat# A22287(33 mM)
ProbeDAPISigma AldrichCat# D9542(500 ng/mL)
ProbeCellTracker Green CMFDA DyeThermo Fisher ScientificCat# C7025(500 ng/mL)
ProbeMitoTracker Deep Red FMInvitrogenCat# M22426(500 ng/mL)
IFFluorescence Mounting MediumAgilent TechnologiesCat# S302380‑2
IFAntibody diluent, Background reducingDAKO, Agilent TechnologiesCat# S302283-2
SoftwareImageJImageJhttp://www.imagej.nih.gov/ij
SoftwarePrism (v8.0.0)GraphPadhttp://www.graphpad.com
SoftwarePhotoshop CS3Adobehttp://www.adobe.com
SoftwareAdobe Illustrator CS5Adobehttps://www.adobe.com/fr/products/illustrator.html
SoftwareHuygens Professional Version 16.10Scientific Volume Imaginghttps://svi.nl/HuygensProfessional
SoftwareFACS DIVABD Biosciencehttp://www.bdbiosciences.com/
SoftwareFlowJo_v10FlowJohttps://www.flowjo.com/
SoftwareFCS Express V3DeNovo Softwarehttp://www.denovosoftware.com
SoftwareImage LabBio‑Rad Laboratorieshttp://www.bio‑rad.com
SoftwarePannoramic Viewer3DHISTECHhttps://www.3dhistech.com/pannoramic_viewer

Additional files

Supplementary file 1

Clinical data of NHPs (Table S1) and histopathological scoring of lung lesions in NHPs (Table S2).

https://cdn.elifesciences.org/articles/52535/elife-52535-supp1-v2.docx
Transparent reporting form
https://cdn.elifesciences.org/articles/52535/elife-52535-transrepform-v2.docx

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)