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: Cell Death & Disease

Article Title: M2 macrophage-mediated interleukin-4 signalling induces myofibroblast phenotype during the progression of benign prostatic hyperplasia

doi: 10.1038/s41419-018-0744-1

Figure Lengend Snippet: a Representative CD68 immunohistochemical staining images in the early-progressed BPH, age-matched prostate and elderly BPH tissues. Scale bar, 200 μm. b Scatter plots showing macrophage clusters in the three analysed groups. The number of macrophage clusters in prostate tissues correlated positively with the stroma-to-epithelium ratio, and percent area densities of α-SMA, collagen I and fibres. c Serial histological sections showing the early-progressed BPH tissues stained for CD68, a macrophage-specific marker, and CD163, an M2 macrophage marker. The distribution of CD68 + and CD163 + cells was almost identical. Scale bar, 200 μm. d Representative immunofluorescence (IF) staining images, showing CD68 (green) and CD163 (red) expression in the infiltrating macrophages in early-progressed BPH tissues. Scale bar, 100 μm. e Quantitative RT-PCR results showing α-SMA, COL1A1 and COL3A1 expression in primary prostate fibroblast (PrPF)-early, PrPF-control and PrPF-old cells co-cultured with THP-1-derived M2 macrophages. Data are shown as relative gene expression compared to that in the respective untreated fibroblasts. f Western blots showing α-SMA and collagen I protein expression in PrPF-early, PrPF-control and PrPF-old cells co-cultured with THP-1-derived M2 macrophages. g Representative IF staining results, showing the co-expression of α-SMA (red) and collagen I (green) following a 48-h co-culture of PrPF-early, PrPF-control and PrPF-old cells with THP-1-derived M2 macrophages. The respective untreated fibroblasts were used as controls. Scale bar, 100 μm. h Solidified collagen-gel shrinkage after the seeding of PrPF-early, PrPF-control and PrPF-old cells, untreated or treated with the conditioned medium. The respective untreated fibroblast samples were used as controls, and all assays were performed in triplicate. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The primary antibodies included mouse monoclonal α-SMA (Abcam, ab7817, 1:100 dilution), CD163 (OriGene, TA506380, 1:200 dilution) and EpCAM (Cell Signaling Technology, #2929, 1:800 dilution) antibodies, and the rabbit primary antibodies included collagen I (Abcam, ab138492, 1:800 dilution), CD68 (Proteintech, 25747-1-AP, 1:400 dilution), IL4R (ImmunoWay, YT2337, 1:400 dilution), TGFβ RI (ImmunoWay, YT4627, 1:400 dilution) and vimentin (Cell Signaling Technology, #5741, 1:100 dilution) antibodies.

Techniques: Immunohistochemical staining, Staining, Marker, Immunofluorescence, Expressing, Quantitative RT-PCR, Control, Cell Culture, Derivative Assay, Gene Expression, Western Blot, Co-Culture Assay

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

Journal: Toxicologic Pathology

Article Title: Biocompatible Solutions: Evaluating the Safety of Repeated Intra-Articular Injections of pMPCylated Liposomes for Knee Osteoarthritis Therapy in Rat Models

doi: 10.1177/01926233241271400

Figure Lengend Snippet: Antibodies used for the immunofluorescence study.

Article Snippet: Novus (CD163) , NBP3-07981 (CD163).

Techniques: Immunofluorescence, Marker

Journal: Journal for immunotherapy of cancer

Article Title: IGF1 receptor inhibition amplifies the effects of cancer drugs by autophagy and immune-dependent mechanisms.

doi: 10.1136/jitc-2021-002722

Figure Lengend Snippet: Figure 4 IGF1R signaling correlates with immunosuppressive markers and decreased survival in breast cancer. (A–I) The expression of CD163, FOXP3 and CD8 as well as the phosphorylation of IGF1R and the dot formation of LC3 were quantified in paraffin-embedded biopsies obtained from 49 triple-negative breast cancer patients by ImageJ after staining with specific antibodies. Representative images of phosphorylated-IGF1R, dotted LC3B, and CD163, FOXP3 and CD8 expression are shown in A. The scale bar indicates 100 μm. Correlation analyses (determined by the Spearman method) of the analyzed parameters for each patient are depicted in B–E. (F–I) Kaplan-Meier survival analysis of patients with biomarker-positive and biomarker- negative immunohistochemistry staining. IGF1R, insulin-like growth factor-1 receptor.

Article Snippet: Sections were dehydrated and antigenic epitopes were retrieved using a 10 mM citrate buffer and microwaving for 10 min. Specimen were then incubated with for LC3B (#3868, 1:2000, Cell Signaling Technology), phospho- IGF1R (#orb503127, 1:100, Biorbyt, UK), CD163 (#93498, 1:500, Cell Signaling Technology), Foxp3 ((#98377, 1:100, Cell Signaling Technology), CD8 (#70306, 1:200, Cell Signaling Technology).

Techniques: Expressing, Phospho-proteomics, Staining, Biomarker Discovery, Immunohistochemistry