Journal: The Journal of Immunology Author Choice

Article Title: Cold Mechanical Isolation of Placental Macrophages as a Method to Limit Procedure-Induced Activation of Macrophages

doi: 10.4049/jimmunol.2300379

Figure Lengend Snippet: Abs used for flow cytometry and FACS

Article Snippet: Ab (label) Clone Source Catalog No. Anti-human CD14 (PE) 61D3 eBioscience 12-0149-41 Anti-human CD45 (FITC) HI30 BioLegend 304006 Anti-human CD80 (PE-Cy7) L307.4 BD Biosciences 561135 Anti-human CD86 (BB7000) 2331 (FUN-1) BD Biosciences 566474 Anti-human CD163 (BV421) GHI/61 BD Biosciences 566277 Anti-human CD206 (allophycocyanin) 19.2 eBioscience 17-2069-41 Open in a separate window Abs used for flow cytometry and FACS Open in a separate window FIGURE 3.

Techniques: Flow Cytometry

Journal: The Journal of Immunology Author Choice

Article Title: Cold Mechanical Isolation of Placental Macrophages as a Method to Limit Procedure-Induced Activation of Macrophages

doi: 10.4049/jimmunol.2300379

Figure Lengend Snippet: Gating strategy to define and isolate macrophages by flow cytometry and FACS. (A) Representative images of the gating strategy to sort single, viable macrophages from placental tissue based on the expression of CD45 and CD14. (B) Representative flow cytometric gating strategy to define CD45+ leukocytes. (C) Representative flow cytometric gating strategy to explore the expression of the surface markers CD80, CD86, CD163, and CD206 on CD14+ macrophages.

Article Snippet: Ab (label) Clone Source Catalog No. Anti-human CD14 (PE) 61D3 eBioscience 12-0149-41 Anti-human CD45 (FITC) HI30 BioLegend 304006 Anti-human CD80 (PE-Cy7) L307.4 BD Biosciences 561135 Anti-human CD86 (BB7000) 2331 (FUN-1) BD Biosciences 566474 Anti-human CD163 (BV421) GHI/61 BD Biosciences 566277 Anti-human CD206 (allophycocyanin) 19.2 eBioscience 17-2069-41 Open in a separate window Abs used for flow cytometry and FACS Open in a separate window FIGURE 3.

Techniques: Flow Cytometry, Expressing

Journal: The Journal of Immunology Author Choice

Article Title: Cold Mechanical Isolation of Placental Macrophages as a Method to Limit Procedure-Induced Activation of Macrophages

doi: 10.4049/jimmunol.2300379

Figure Lengend Snippet: Comparison of CD163high cells between the cold and warm protocol. The expression of CD163 within the CD163highCD206high and CD163lowCD206high population was compared between placentas processed with the cold mechanical and warm enzymatic protocol (n = 10) with the Wilcoxon signed-rank test. (A) CD163 expression given as median in the CD206highCD163low population. (B) CD163 expression given as median in the CD206highCD163high population. Data are presented in a boxplot showing median and 1st and 3rd IQR. Whiskers indicate 5th to 95th percentiles. Light gray represents cold mechanical isolation; dark gray represents warm enzymatic isolation. *p < 0.05.

Article Snippet: Ab (label) Clone Source Catalog No. Anti-human CD14 (PE) 61D3 eBioscience 12-0149-41 Anti-human CD45 (FITC) HI30 BioLegend 304006 Anti-human CD80 (PE-Cy7) L307.4 BD Biosciences 561135 Anti-human CD86 (BB7000) 2331 (FUN-1) BD Biosciences 566474 Anti-human CD163 (BV421) GHI/61 BD Biosciences 566277 Anti-human CD206 (allophycocyanin) 19.2 eBioscience 17-2069-41 Open in a separate window Abs used for flow cytometry and FACS Open in a separate window FIGURE 3.

Techniques: Comparison, Expressing, Isolation

Journal: Acta biomaterialia

Article Title: A subtype specific probe for targeted magnetic resonance imaging of M2 tumor-associated macrophages in brain tumors.

doi: 10.1016/j.actbio.2025.01.003

Figure Lengend Snippet: Fig. 6. Immunohistochemical and FACS validations of M2pep-uIONP targeting M2 TAMs. Confocal microscopic images of tumor tissues collected from mice at 24 h after i.v. injection of probes show (A) Cy7-M2pep-uIONP were highly co-localizing with CD68- and CD163-double positive M2 TAM, revealing the targeting specificity. In comparison, the non-targeted control probe Cy7-scM2pep-uIONP and Cy7-Ferumoxytol not only exhibited much less tumoral accumulation, but also were not co-localized with M2 TAM. Scale bars: 37 µm. (B) Flow cytometric zebra plots and (C) the corresponding quantification of CD68- and CD163-double positive M2 TAM targeted by FITC-M2pep-uIONP, FITC-scM2pep-uIONP, and FITC-Ferumoxytol. The gating strategy involves sorting CD163+ M2 TAM population in live cells dissociated from mouse cerebrums by gating on CD68+ pan-macrophage populations, followed by further gating to assess the FITC-labeled nanoparticles.

Article Snippet: Rabbit polyclonal anti-CD163 antibody was purchased from Boster Biological Technology (Pleasanton, CA, USA).

Techniques: Immunohistochemical staining, Injection, Comparison, Control, Labeling

Journal: Journal of the Endocrine Society

Article Title: Transgender Women With Suppressed Testosterone Display Lower Burden of Coronary Disease Than Matched Cisgender Men

doi: 10.1210/jendso/bvae120

Figure Lengend Snippet: Biomarker concentrations

Article Snippet: Soluble CD14 (sCD14, R and D Systems catalog No. DC140, RRID:AB_3095885), soluble CD163 (sCD163, R and D Systems catalog No. DC1630, RRID:AB_3096052), interleukin-6 (IL-6, R and D Systems catalog No. HS600C, RRID:AB_2893335), fatty acid-binding protein-4 (FABP-4, (ImmunoDiagnostics catalog No. 31030, RRID:AB_2933962), high-molecular-weight adiponectin (R and D Systems catalog No. DRP300, RRID:AB_2783020), insulin (R and D Systems catalog No. DINS00, RRID:AB_3073852), endothelin-1 (ET-1, R and D Systems catalog No. DET100, RRID:AB_3099470), and vascular cell adhesion molecule-1 (VCAM-1, R and D Systems catalog No. DVC00, RRID:AB_2941366) were analyzed by Quantikine ELISA (enzyme-linked immunosorbent assay), and soluble tumor necrosis factor receptor type (sTNFR, R and D Systems catalog No. LUCAM726, RRID:AB_3099473) I/II, interleukin-8 (IL-8, R and D Systems catalog No. DY208, RRID:AB_2892143), and plasminogen activator inhibitor-1 (PAI-1, R and D Systems catalog No. LOBM1786, RRID:AB_3099472) were measured by Luminex assay.

Techniques: Biomarker Discovery

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: IL-13–regulated Macrophage Polarization during Granuloma Formation in an In Vitro Human Sarcoidosis Model

doi: 10.1165/rcmb.2018-0053oc

Figure Lengend Snippet: Figure 5: Immunofluorescence imaging of granuloma-like structures demonstrates persistent macrophage CD163 expression on sarcoidosis PBMCs following PPD stimulation. Representative photomicrographs of immunostained PBMCs in granuloma-like structures 7 days after bead treatment using

Article Snippet: Copyright © 2018 by the American Thoracic Society Alternatively, following washing and blocking, some coverslips were incubated with 0.5 μg/ml CD163 [unlabeled, rabbit, anti-human CD163 mAb (K20-T, IgG) from Novus Biologicals, LLC (Littleton, CO)] or the matching isotype control antibody [Abcam, (Cambridge, MA)] at room temperature for 1 hour.

Techniques: Immunofluorescence, Imaging, Expressing

Journal: Human reproduction (Oxford, England)

Article Title: Distribution of decidual natural killer cells and macrophages in the neighbourhood of the trophoblast invasion front: a quantitative evaluation.

doi: 10.1093/humrep/det353

Figure Lengend Snippet: Figure 5 Distribution of dMph. (a) Cryostat section from a cell column at a first trimester placentation site double-stained with an anti-CD163 antibody (red) and an anti-HLA-G antibody (dark blue) as a sample image. Scale bar ¼ 100 mm. (b) Box plots of normalized areal cell densities s of dMph at first trimester placentation sites according to Equation (15), in five trophoblast neighbourhood rings according to Equation (17), are depicted. The dashed line s ¼ 1 represents the average areal cell density. #P ¼ 0.05, ##P ¼ 0.01 compared with 1 by Student’s t-tests (H0:m ¼ 1). ANOVA analysis showed a high significance,P ≤0.01.*P ¼ 0.05,**P ¼ 0.01ofpair-wiseTukeyhonestsignificantdifferenceposthoctests(s ¼ 1).(c)DistributionofdMphinco-culturesof first trimester decidua parietalis with first trimester villous explants using cryostat sections from co-cultures double-stained with an anti-CD163 antibody and an anti-HLA-G antibody. Box plots of normalized areal cell densities s. #P ¼ 0.05, ##P ¼ 0.01 compared with 1 by Student’s t-tests (H0:m ¼ 1). ANOVA analysis showed a high significance, P ≤0.01. **P ¼ 0.01 of pair-wise Tukey honest significant difference post hoc tests (s ¼ 1). Open circle is an outlier. (d) Distribution of dMph in first trimester decidua parietalis determined by using cryostat sections from tissue fragments stained with an anti-CD163 antibody. Box plots of normalized areal cell densities s. P . 0.05 by ANOVA test. Filled circle is an extreme value.

Article Snippet: The following antibodies were used: † monoclonal mouse anti-human CD56 (clone 123C3; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), dilution: 1:400, final concentration: 0.5 mg/ml; † monoclonal mouse anti-human CD163 (clone 5C6-FAT; Acris Antibodies GmbH, Hiddenhausen, Germany), dilution: 1:500, final concentration: 0.4 mg/ml; † monoclonal mouse anti-human HLA-G (clone 4H84; Exbio, Praha, Czech Republic; supplied by Eubio, Vienna, Austria), dilution: 1:500, final concentration: 2 mg/ml.

Techniques: Staining