Journal: Journal of Translational Medicine

Article Title: Lupus serum IgG induces microglia activation through Fc fragment dependent way and modulated by B-cell activating factor

doi: 10.1186/s12967-019-02175-0

Figure Lengend Snippet: Flow cytometric analysis of microglia surface markers in SLE-serum, healthy-serum and ACSF-treated mice (n = 16, 14 and 10). a Cells were defined by gate R1 to eliminate unwanted events, such as cell debris. b Microglias were identified by their CD45 low CD11b + phenotype (R2). c Isotype for MHCII was used to define the gating of M1 cells. In this population, SLE-serum led to a significantly increased percentage of MHC-II + CD45 low CD11b + cells. d Isotype for CD206 was used to define the gating of M2 cells. In contrast, percentage of CD206 + was not significantly changed by SLE-serum treatment in the CD45 low CD11b + cells. Bars represent the mean ± SEM of mice groups received injection of SLE-serum, healthy-serum or ACSF. Pictures of c and d show representative histogram and flow dot plots of MHC-II and CD206 expression, respectively, in the CD45 low CD11b + population. *p < 0.05; **p < 0.01

Article Snippet: Then, the following antibodies were used for mouse microglia surface staining: PE-Cy7 rat anti-mouse CD45 (#130-110-799, MiltenyiBiotec, BergischGladbach, Germany), APC-Cy7 rat anti-mouse CD11b (#130-109-366, MiltenyiBiotec, BergischGladbach, Germany), FITC rat anti-mouse MHCII (#11-5322-81, Invirogen, Carlsbad, USA), isotype for MHCII (#11-4031-81, Invirogen, Carlsbad, USA), Percp-cy5.5 rat anti-mouse CD206 (#141715, BioLegend, San Diego, USA) and isotype for CD206 (#400531, BioLegend, San Diego, USA).

Techniques: Injection, Expressing

Journal: PLoS ONE

Article Title: Simvastatin Reduces Endotoxin-Induced Acute Lung Injury by Decreasing Neutrophil Recruitment and Radical Formation

doi: 10.1371/journal.pone.0038917

Figure Lengend Snippet: Isolated human neutrophils were pre-treated with Simvastatin (3 hours, 1 or 10 µM) and then activated with fMLP. MFI of surface expression of CD11b (A), CD29 (B) and FPRL1 (C) as measured by flow cytometry after staining with directly conjugated antibodies (n = 3–6; repetition = 4). D+E In vivo degranulation after LPS inhalation. Mice were challenged with LPS via inhalation and sacrificed 4 hours later. After isolation of neutrophils from the blood and the lung, MFI of surface expression of CD11b and CD29 as measured by flow cytometry after staining with directly conjugated antibodies (n = 4–6). Statistical significance was tested using one way ANOVA with Newman-Keuls Multiple Comparison test. * indicates significant difference in comparison to the fMLP or the LPS group.

Article Snippet: Cell pellets were labeled with PerCP-Cy5.5 anti-mouse Ly-6G, PE anti-mouse CD115, APC-Cy7 anti-mouse CD45 und APC anti-Mouse CD11b (all eBioscience) and FITC anti-mouse CD29 (Miltenyi Biotec).

Techniques: Isolation, Expressing, Flow Cytometry, Staining, In Vivo, Comparison

Journal: bioRxiv

Article Title: Unbiased metastatic niche-labeling identifies estrogen receptor-positive macrophages as a barrier of T cell infiltration during bone colonization

doi: 10.1101/2024.05.07.593016

Figure Lengend Snippet: (A) Cell types, tissue origin, and biotin groups in SAMENT single-cell dataset. (B) GSVA analysis comparing biotin-positive NK cells to biotin-negative NK cells highlights potential regulations exerted by NK cells on macrophages. Biotin-positive NK cells appear to modulate the tumor microenvironment by enhancing chemotaxis and inhibiting both aging and apoptosis in macrophages. This suggests that upon encountering tumors, reprogrammed NK cells indirectly contribute to chronic inflammation and promote the survival of tumor-associated macrophages. (C) Top enriched pathways in biotin-positive immature B cells in bones compared with other tissues. (D) Top enriched pathways in monocyte and neutrophil precursors in lungs compared with other tissues. (E) Top enriched pathways in biotin-positive granulocyte-macrophage progenitors (GMPs) in bone compared with biotin-negative populations. (F) Validation of published tissue-specific macrophage signatures was conducted in the SAMENT dataset. Reported Microglia signatures include Tmem119 + , Trem2 + , Fcrls + , Slc2a5 + , P2ry12 + , and Cx3cr1 + . Alveolars signatures consist of Cd163 − , Cd11b − , Cd206 int , Spp1 + , SiglecF + and Cd11c + ; Osteoclasts signatures are characterized by Arg1 + , Spp1 + , Mmp9 + , and vacuolar [H+]-ATPase + ; Kupffers cell signatures include CD11b low , F4/80 high , and Clec4F + ; Monocyte-derived macrophage signatures , encompass CD11b + , F4/80 int , Ly6C + and CSF1R + . In our dataset, Mφ_c10 corresponds to Microglia, Mφ_c7 corresponds Alveolar macrophages, and Mφ_c5 corresponds to osteoclasts. Monocyte-derived macrophages are distributed across all macrophage subclusters, while we did not identify specific clusters for Kupffer cells. These findings support the observations depicted in .

Article Snippet: The antibodies used in this study included: Immune cell panel, CD45-VF450 (Cytek® Biosciences,75–0451), CD11b-APC/Cy7 (Cytek® Biosciences, 25–0112), Ly6G-Percp/Cy5.5 (Cytek® Biosciences, 65–1276), Ly6C-PE/Cy7 (BioLegend,128018), F4/80-BV510 (BioLegend,123135), B220-APC/Cy7 (Cytek® Biosciences, 25-0452), CD3e-Percp/Cy5.5 (Cytek® Biosciences,65-0031), CD4-PE/Cy7 (Cytek® Biosciences,60-0041), CD8a-BV711 (BD Biosciences,752634), PD-1-BV605 (BioLegend,135219), Biotin-APC (Miltenyi Biotec,130-113-288) or Biotin-PE(Miltenyi Biotec,130-113-291); Stromal cell panel, CD45-BV605 (BioLegend,103140), Ter119-BV605 (BioLegend,116239), CD31-APC/Cy7 (BioLegend,102440), Sca-1-Percp/Cy5.5 (eBioscience,45–5981-82), CD51-BV421 (BD Biosciences, 740062), CD140α-PE/Cy7 (BioLegend, 135912), Biotin-APC (Miltenyi Biotec,130-113-288) or Biotin-PE(Miltenyi Biotec,130-113-291) Flow cytometry analysis was conducted using a BD LSRFortessa flow cytometer and data were further analyzed with FlowJo software.

Techniques: Chemotaxis Assay, Biomarker Discovery, Derivative Assay