Journal: PLoS ONE

Article Title: CD200R/CD200 Inhibits Osteoclastogenesis: New Mechanism of Osteoclast Control by Mesenchymal Stem Cells in Human

doi: 10.1371/journal.pone.0072831

Figure Lengend Snippet: PBMC adherent cells were co-cultured with MSCs in presence of M-CSF for 48 hr. RANKL was added after 48 hr and half of the medium was changed every 3 days. (A) After 21 days of culture, actin rings were stained with phalloidin-Alexa 540 and nuclei with DAPI. (B) Osteoclast precursors were cultivated with MSCs, CD200 + MSCs or CD200 – MSCs for 21 days. Tartrate-resistant acid phosphatase (TRAP) assay was performed and area of TRAP-positive cells was measured. Data are mean±SD. *, P<0.05. (C) Effect of MSCs on bone resorption activity of osteoclasts. White arrows represent bone resorption pits. The results of 1 experiment out of 3 are presented.

Article Snippet: Recombinant human CD200 Fc chimera protein (rCD200) was from R&D Systems (Minneapolis, MN, USA).

Techniques: Cell Culture, Staining, TRAP Assay, Activity Assay

Journal: Stem cells (Dayton, Ohio)

Article Title: CD200 Expression Marks a Population of Quiescent Limbal Epithelial Stem Cells with Holoclone Forming Ability.

doi: 10.1002/stem.2903

Figure Lengend Snippet: Figure 2. CD200 expression in human and mouse cornea in vivo and during ex vivo expansion of human limbal epithelial cells. (A): Quantification of CD200 expression through different passages of limbal epithelial cells by flow cytometry. Values represent mean SEM, n = 3–10 (n, number of biological replicates), *, p < .05. (B): Quantification of CD200 expression during calcium induced differentiation of limbal epithelial cells by flow cytometry. Values represent mean SEM, n = 3, *, p < .05. (C): Immunohistochemical staining of human corneal tissue paraffin sections for ΔNp63 and CD200 within the central cornea and limbus. Nuclei are shown by Hoechst counter stain- ing. Scale bars 20 μm. (D): Immunohistochemical staining of murine corneal tissue cryosections for CK15, ΔNp63, and CD200 within the central cornea and limbus. Nuclei are shown by DAPI counter staining. The dashed line indicates the stromal-epithelial junction. Red arrows point at limbal region. Scale bars 20 μm. (E): Immunohistochemical staining of limbal epithelial cell colonies in vitro for CD200 and ΔNp63. Blue arrow points CD200+ cells. Nuclei are shown by Hoechst counter staining. Scale bar 50 μm. (F): Immunohistochemical stain- ing of limbal epithelial cell colonies in vitro for CD200 and Ki67. Red arrows point to CD200+ Ki67+ cells; orange arrows point to CD200+Ki67−cells. Nuclei are shown by Hoechst counter staining. Scale bar 50 μm. Abbreviations: ep, epithelium; st, stroma.

Article Snippet: The following primary antibodies were used at the indicated dilutions: anti CD109 (sc-271085, Santa Cruz, USA, 1:200), anti-human CD200 (329201, BioLegend, USA, 1:200), anti-mouse CD200 (AF3355, Novus Biologicals, USA, 1:100), anti p63 delta (NBP2-29467, Novus Biologicals, USA, 1:200), anti-cytokeratin 15 (ab52816, Abcam, Cambridge, UK, 1:100), and anti Ki67 antibody (ab15580, Abcam, UK, 1:100).

Techniques: Expressing, In Vivo, Ex Vivo, Cytometry, Immunohistochemical staining, Staining, In Vitro

Journal: Stem cells (Dayton, Ohio)

Article Title: CD200 Expression Marks a Population of Quiescent Limbal Epithelial Stem Cells with Holoclone Forming Ability.

doi: 10.1002/stem.2903

Figure Lengend Snippet: Figure 3. Colony forming efficiency and proliferative potential of sorted CD200 positive and negative population. (A): Pie chart showing the distribution of formed and aborted colonies in CD200+ population. (B): Comparison of colony forming efficiencies of CD200+ and CD200−cells. Values represent mean SEM, n = 3 (n, number of biological replicates). (C): Pie chart showing the distribution of formed and aborted colonies in CD200−population. Values represent mean SEM, n = 3. (D): Microscopic and macroscopic appearances of colo- nies formed by CD200+ cells. Scale bars 100 μm. (E): Microscopic and macroscopic appearances of colonies formed by CD200−cells. Scale bars 100 μm. (F): BrdU cell proliferation assay of CD200 negative and positive limbal epithelial cell population after 1- and 8-hours incuba- tion with BrdU. Values represent mean SEM, n = 3.(G): Quantification of cells in the S phase of the cell cycle in CD200+ and CD200−

Article Snippet: The following primary antibodies were used at the indicated dilutions: anti CD109 (sc-271085, Santa Cruz, USA, 1:200), anti-human CD200 (329201, BioLegend, USA, 1:200), anti-mouse CD200 (AF3355, Novus Biologicals, USA, 1:100), anti p63 delta (NBP2-29467, Novus Biologicals, USA, 1:200), anti-cytokeratin 15 (ab52816, Abcam, Cambridge, UK, 1:100), and anti Ki67 antibody (ab15580, Abcam, UK, 1:100).

Techniques: Comparison, BrdU Cell Proliferation Assay

Journal: Stem cells (Dayton, Ohio)

Article Title: CD200 Expression Marks a Population of Quiescent Limbal Epithelial Stem Cells with Holoclone Forming Ability.

doi: 10.1002/stem.2903

Figure Lengend Snippet: Figure 5. CD200 knockdown and its effect on clonal ability of limbal epithelial cells. (A): Quantitative reverse transcriptase poly- merase chain reaction expression data for control siRNA versus CD200 siRNA treated limbal epithelial cells. Values represent mean SEM, n = 3 (n, number of biological replicates), *, p < .05. (B): Pie chart showing distribution of paraclones, meroclones, and holoclones formed by control siRNA treated cells and (C) CD200 siRNA treated cells. (D): Representative images of colonies formed in control and CD200 siRNA group, with 500 or 1,000 cells seeded per well. Abbreviation: siRNA, small interfering RNA.

Article Snippet: The following primary antibodies were used at the indicated dilutions: anti CD109 (sc-271085, Santa Cruz, USA, 1:200), anti-human CD200 (329201, BioLegend, USA, 1:200), anti-mouse CD200 (AF3355, Novus Biologicals, USA, 1:100), anti p63 delta (NBP2-29467, Novus Biologicals, USA, 1:200), anti-cytokeratin 15 (ab52816, Abcam, Cambridge, UK, 1:100), and anti Ki67 antibody (ab15580, Abcam, UK, 1:100).

Techniques: Knockdown, Reverse Transcription, Expressing, Control, Small Interfering RNA

Journal: Stem cells (Dayton, Ohio)

Article Title: CD200 Expression Marks a Population of Quiescent Limbal Epithelial Stem Cells with Holoclone Forming Ability.

doi: 10.1002/stem.2903

Figure Lengend Snippet: Figure 4. Expression of putative limbal stem cell and corneal epi- thelial cell markers in the sorted CD109 and CD200 positive and negative cell populations. (A): Quantitative reverse transcriptase polymerase chain reaction expression data for CD109+ limbal epi- thelial cell population versus CD109−limbal epithelial cell popula- tion represented by the red line (value 1). Values represent mean SEM, n = 3 (n, number of biological replicates), *, p < .05; **, p < .01; ***, p < .001. (B): Quantitative reverse transcriptase polymerase chain reaction expression data for CD200+ limbal epi- thelial cell population versus CD200−limbal epithelial cell popula- tion represented by the red line (value 1). Values represent mean SEM, n = 3, *, p < .05; **, p < .01; ***, p < .001.

Article Snippet: The following primary antibodies were used at the indicated dilutions: anti CD109 (sc-271085, Santa Cruz, USA, 1:200), anti-human CD200 (329201, BioLegend, USA, 1:200), anti-mouse CD200 (AF3355, Novus Biologicals, USA, 1:100), anti p63 delta (NBP2-29467, Novus Biologicals, USA, 1:200), anti-cytokeratin 15 (ab52816, Abcam, Cambridge, UK, 1:100), and anti Ki67 antibody (ab15580, Abcam, UK, 1:100).

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction

Journal: ERJ Open Research

Article Title: Distinct immune regulatory receptor profiles linked to altered monocyte subsets in sarcoidosis

doi: 10.1183/23120541.00804-2020

Figure Lengend Snippet: Distribution of the regulatory receptor CD200R and its ligand CD200L in sarcoidosis granulomas: transbronchial lung biopsy samples from two patients with sarcoidosis stained for a) CD200L and b) CD200R, with respective isotype control antibody staining in c) and d). f: fibroblasts; h: histiocytes (macrophages). Original magnification ×100.

Article Snippet: Antibodies were a polyclonal goat CD200L antibody (AF2724; R&D Systems, Abingdon, UK), normal goat IgG control (AB-108-C; R&D Systems), mouse monoclonal IgG1 anti-human CD200R (OX108, MCA 2282T; AbD Serotec/Bio-Rad, Kidlington, UK) and mouse monoclonal control IgG1 antibody (MOPC-21, 400101; Biolegend, London, UK).

Techniques: Staining, Control

Journal: bioRxiv

Article Title: Reduced cutaneous CD200:CD200R1 signalling in psoriasis enhances neutrophil recruitment to skin

doi: 10.1101/2022.04.01.486720

Figure Lengend Snippet: A . CD200 QPCR on healthy (NN), non-lesional (PN) and lesional (PP) psoriasis skin relative to the mean NN value. B . Flow cytometry showing CD200 (black line) and isotype control (grey filled histograms) on NN and PN skin. C-E . Immunohistochemistry showing: C . CD200 in NN and PN skin with a secondary only control, and blockade of the anti-CD200 signal by prior incubation with soluble CD200, D . pDok1 co-staining with CD200 or CD200R1 in NN skin, E . pDok1 in NN and PN skin. Bar charts show all data (n = 4-6), Mean and SD shown. A was analysed by Ordinary ANOVA and Dunnett’s test. B, C and E were analysed by Mann Whitney test. * indicates p < .05.

Article Snippet: On occasion, anti-human CD200 was pre-incubated with a 1.5-fold molar amount of human CD200Fc (R&D Systems) prior to staining, to check specificity.

Techniques: Flow Cytometry, Control, Immunohistochemistry, Incubation, Staining, MANN-WHITNEY

Journal: Scientific Reports

Article Title: CD200-positive cancer associated fibroblasts augment the sensitivity of Epidermal Growth Factor Receptor mutation-positive lung adenocarcinomas to EGFR Tyrosine kinase inhibitors

doi: 10.1038/srep46662

Figure Lengend Snippet: ( A ) Heat map plot of CAF608, CAF621, CAF608-clone 4 and CAF608-clone 6. ( B ) Venn diagram showing the number of common genes between the sets of genes whose expression in either the (heterogeneous) CAF608 population or the CAF608-clone 4 was at least four times higher than the expression in CAF621 cells or the CAF608-clone 6, respectively. ( C ) Venn diagram showing the 112 genes after enrichment for two gene ontology terms, “signal” and “cell membrane”. ( D ) Relative NRCAM and CD200 mRNA levels of CAF608, CAF621, CAF608-clone 4 and CAF608-clone 6 cells as determined by RT-PCR assays.

Article Snippet: Tissue sections were stained with antibody recognizing human CD200 (R&D systems, Minneapolis, MN, USA) or a monoclonal mouse antibody recognizing human smooth muscle actin (DAKO, Carpenteria, CA, USA).

Techniques: Expressing, Membrane, Reverse Transcription Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: CD200-positive cancer associated fibroblasts augment the sensitivity of Epidermal Growth Factor Receptor mutation-positive lung adenocarcinomas to EGFR Tyrosine kinase inhibitors

doi: 10.1038/srep46662

Figure Lengend Snippet: ( A ) Relative CD200 mRNA levels of CAF608 cultures transfected with either control vector (sh Luc), sh CD200-1 or -2. ( B ) CD200 protein levels of CAF608, CAF608 sh Luc, sh CD200-1 and -2 cells. ( C ) The percent of control of viable PC9-mRFP cells after gefitinib treatment of cocultures consisting of PC9-mRFP and CAF608 sh Luc, CAF608 sh CD200 -1 or CAF608 sh CD200-2 cells. ( D ) Numbers of CAF608 cells transfected with sh Luc, sh CD200-1 or -2 after gefitinib administration. ( E ) Infection efficacy of lentiviruses as determined by FACS analysis (X axis: FL-4H, Y axis: FSC-H). ( F ) The percent of control of viable PC9-mRFP cells observed after gefitinib treatment of PC9-mRFP cultured alone or cocultured with either CAF621-Control or CAF621-CD200 + (overexpressing of CD200) cells.

Article Snippet: Tissue sections were stained with antibody recognizing human CD200 (R&D systems, Minneapolis, MN, USA) or a monoclonal mouse antibody recognizing human smooth muscle actin (DAKO, Carpenteria, CA, USA).

Techniques: Transfection, Plasmid Preparation, Infection, Cell Culture

Journal: Scientific Reports

Article Title: CD200-positive cancer associated fibroblasts augment the sensitivity of Epidermal Growth Factor Receptor mutation-positive lung adenocarcinomas to EGFR Tyrosine kinase inhibitors

doi: 10.1038/srep46662

Figure Lengend Snippet: ( A ) Immunohistochemical findings of α-SMA-positive/CD200-positive and α-SMA-positive/CD200-negative CAFs. ( B ) Recurrence free survival of EGFR mutation-positive lung adenocarcinoma patients with CD200-positive or CD200-negative CAFs. ( C ) Progression free survival of gefitinib-treated EGFR mutation-positive lung adenocarcinoma patients with CD200-positive or CD200-negative CAFs.

Article Snippet: Tissue sections were stained with antibody recognizing human CD200 (R&D systems, Minneapolis, MN, USA) or a monoclonal mouse antibody recognizing human smooth muscle actin (DAKO, Carpenteria, CA, USA).

Techniques: Immunohistochemical staining, Mutagenesis

Journal: Stroke

Article Title: Neuronal CD200 Signaling Is Protective in the Acute Phase of Ischemic Stroke.

doi: 10.1161/STROKEAHA.120.032374

Figure Lengend Snippet: Figure 1. Effects of neuronal CD200 signaling on stroke outcomes and neuronal apoptosis at 3 days after stroke. (A) Representative images of brain slices stained with cresyl violet (CV) and quantification of brain infarct volumes, NDS, and corner test scores in TMX vs. VEH mice. (B) TUNEL labelled cell apoptosis in peri-infarct area of TMX and VEH treated mice and the quantification of percent of TUNEL positive cells in ipsilateral hemispheres Each dot represents the average of % apoptotic cells from eight 20× fields/animal in the peri-infarct area. N=3 for sham and 6–7 for stroke per group; *P < 0.05.

Article Snippet: Determination of CD200 and cytokine Levels The levels of CD200 in mouse brain, plasma and in the supernatants of primary neuronal culture were measured by a sandwich ELISA kit according to manufacture instruction ((# EK1185, Mouse CD200 PicoKine ELISA Kit, Boster Biological, Pleasanton, CA, USA.).

Techniques: Staining, TUNEL Assay

Journal: Stroke

Article Title: Neuronal CD200 Signaling Is Protective in the Acute Phase of Ischemic Stroke.

doi: 10.1161/STROKEAHA.120.032374

Figure Lengend Snippet: Figure 4. Plasma cytokine levels after ischemic injury. Pro-inflammatory (TNFα, IL-1β and IL-6) and anti-inflammatory (IL-10 and IL-4) cytokine levels (pg/mL) were measured in the plasma of VEH and TMX treated sham/stroke mice at 3d (A) and 7d (B). N=5–6 sham and 6–7 stroke animals per group; *P < 0.05. (C) CD200 levels in naïve brains (left), in the plasma (middle) of TMX and VEH treated mice, and in neuronal culture medium after OGD (right). N=5–6 sham and 6–8 stroke animals per group; *P < 0.05. For neuronal culture medium, N=4 independent OGD experiments; *P < 0.05.

Article Snippet: Determination of CD200 and cytokine Levels The levels of CD200 in mouse brain, plasma and in the supernatants of primary neuronal culture were measured by a sandwich ELISA kit according to manufacture instruction ((# EK1185, Mouse CD200 PicoKine ELISA Kit, Boster Biological, Pleasanton, CA, USA.).

Techniques: Clinical Proteomics

Journal: Stroke

Article Title: Neuronal CD200 Signaling Is Protective in the Acute Phase of Ischemic Stroke.

doi: 10.1161/STROKEAHA.120.032374

Figure Lengend Snippet: Figure 5. Stroke outcomes and flow cytometry in CD200 Fc treated mice. (A) Representative images of brain slices stained with cresyl violet (CV) and quantification of brain infarct volumes, NDS, and corner test scores in CD200Fc vs. IgG treated mice. N=3 for sham and 6–7 for stroke per group; *P < 0.05. (B) Microglial expression of cell membrane markers CD68/CD206 measured as mean fluorescence intensity (MFI). (C) Microglial expression of intracellular markers IL-1β/TNFα. For (B) and (C), n=4/sham group and n=6/stroke group; *P < 0.05.

Article Snippet: Determination of CD200 and cytokine Levels The levels of CD200 in mouse brain, plasma and in the supernatants of primary neuronal culture were measured by a sandwich ELISA kit according to manufacture instruction ((# EK1185, Mouse CD200 PicoKine ELISA Kit, Boster Biological, Pleasanton, CA, USA.).

Techniques: Flow Cytometry, Staining, Expressing, Membrane, Fluorescence

Journal: Stroke

Article Title: Neuronal CD200 Signaling Is Protective in the Acute Phase of Ischemic Stroke.

doi: 10.1161/STROKEAHA.120.032374

Figure Lengend Snippet: Figure 6. Interaction of neuronal CD200 with lymphocytic CD200R1. (A) The purity of CD45+CD3+

Article Snippet: Determination of CD200 and cytokine Levels The levels of CD200 in mouse brain, plasma and in the supernatants of primary neuronal culture were measured by a sandwich ELISA kit according to manufacture instruction ((# EK1185, Mouse CD200 PicoKine ELISA Kit, Boster Biological, Pleasanton, CA, USA.).

Techniques:

Journal: Life

Article Title: Prenatal Immune Challenge Differentiates the Effect of Aripiprazole and Risperidone on CD200–CD200R and CX3CL1–CX3CR1 Dyads and Microglial Polarization: A Study in Organotypic Cortical Cultures

doi: 10.3390/life14060721

Figure Lengend Snippet: Genes (with corresponding catalogue numbers of TaqMan probes) that were determined in control and MIA OCCs under basal conditions and after exposure to aripiprazole (50 and 100 µM) or risperidone (5 µM) and/or lipopolysaccharide (LPS) using qRT-PCR. Gapdh served as the reference gene.

Article Snippet: Cd200 , Rn01646320_m1.

Techniques: Control

Journal: Life

Article Title: Prenatal Immune Challenge Differentiates the Effect of Aripiprazole and Risperidone on CD200–CD200R and CX3CL1–CX3CR1 Dyads and Microglial Polarization: A Study in Organotypic Cortical Cultures

doi: 10.3390/life14060721

Figure Lengend Snippet: The mRNA expression of Cd200r , Cd200 , Cx3cr1 and Cx3cl1 in control and MIA OCCs after treatment with aripiprazole (50 and 100 µM) or risperidone (5 µM) under basal and LPS-stimulated conditions. n = 3–7 in control OCCs and n = 2–6 in MIA OCCs ( Cd200r , Cd200 , Cx3cl1 ), n = 3–8 in control OCCs and n = 2–6 in MIA OCCs ( Cx3cr1 ). The results were calculated as the average fold change and are displayed as the means ± standard errors of the means (SEM). Statistical analysis was performed using factorial ANOVA with Duncan’s post hoc test or planned comparisons via one-way ANOVA (contrast analysis). * p < 0.05 vs. control OCCs + vehicle, # p < 0.05 vs. control OCCs + LPS, ^ p < 0.05 vs. MIA OCCs + vehicle [ 58 ].

Article Snippet: Cd200 , Rn01646320_m1.

Techniques: Expressing, Control