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: 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: Heliyon

Article Title: CD200-CD200R affects cisplatin and paclitaxel sensitivity by regulating cathepsin K-mediated p65 NF-κB signaling in cervical cancer

doi: 10.1016/j.heliyon.2023.e19220

Figure Lengend Snippet: CD200 and CTSK expression in cervical cancer tissues. (A) The GEPIA online website was used to analyze CD200 and CTSK expression in cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) tissues and normal tissues. (B) The correlation of CD200 and CTSK in the CESC. CD200 (C) and CTSK (D) expression in tumor tissues and para-cancerous tissues was measured using immunohistochemistry. *P < 0.05, **P < 0.01. Representative images of CD200 and CTSK expression in tissues are provided in Supplemental and .

Article Snippet: Diluted primary CD200 rabbit antibody (#23451S, 1:200, Cell Signaling Technology, Shanghai, China), CD200R rabbit-antibody (1:400, #DF4715, Affinity, China), Ki-67 rabbit-antibody (1:400, #9027, Cell Signaling Technology) and CTSK rabbit antibody (1:400, #DF6614, Affinity, China) were incubated with the section at 4 °C for overnight.

Techniques: Expressing, Immunohistochemistry

Journal: Heliyon

Article Title: CD200-CD200R affects cisplatin and paclitaxel sensitivity by regulating cathepsin K-mediated p65 NF-κB signaling in cervical cancer

doi: 10.1016/j.heliyon.2023.e19220

Figure Lengend Snippet: Effects of CD200 on the viability and invasion of HeLa and Siha cells. HeLa and Siha cells were stably transfected with 2 μg of control plasmid or PUNO1-hCD200. (A) Cell viabilities were analyzed by CCK-8. (B) Cell invasion was measured by Transwell assay. Scale = 50 μm. The results of CD200 transfection in HeLa and SiHa cells are provided in Supplemental and . NS: no significance.

Article Snippet: Diluted primary CD200 rabbit antibody (#23451S, 1:200, Cell Signaling Technology, Shanghai, China), CD200R rabbit-antibody (1:400, #DF4715, Affinity, China), Ki-67 rabbit-antibody (1:400, #9027, Cell Signaling Technology) and CTSK rabbit antibody (1:400, #DF6614, Affinity, China) were incubated with the section at 4 °C for overnight.

Techniques: Stable Transfection, Transfection, Control, Plasmid Preparation, CCK-8 Assay, Transwell Assay

Journal: Heliyon

Article Title: CD200-CD200R affects cisplatin and paclitaxel sensitivity by regulating cathepsin K-mediated p65 NF-κB signaling in cervical cancer

doi: 10.1016/j.heliyon.2023.e19220

Figure Lengend Snippet: CD200-CD200R regulated the CTSK-mediated p65 NF-κB pathway in cervical cancer cells. Cancer cells were cocultured without or with M2 macrophage-like THP-1 cells. Expression levels of CTSK and p-p65 NF-κB/p65 NF-κB protein in HeLa (A) and Siha (B) were measured by Western blotting. The full length of blots was provided in Supplemental materials. *P < 0.05, **P < 0.01.

Article Snippet: Diluted primary CD200 rabbit antibody (#23451S, 1:200, Cell Signaling Technology, Shanghai, China), CD200R rabbit-antibody (1:400, #DF4715, Affinity, China), Ki-67 rabbit-antibody (1:400, #9027, Cell Signaling Technology) and CTSK rabbit antibody (1:400, #DF6614, Affinity, China) were incubated with the section at 4 °C for overnight.

Techniques: Expressing, Western Blot

Journal: Heliyon

Article Title: CD200-CD200R affects cisplatin and paclitaxel sensitivity by regulating cathepsin K-mediated p65 NF-κB signaling in cervical cancer

doi: 10.1016/j.heliyon.2023.e19220

Figure Lengend Snippet: CD200-CD200R is involved in the response to cisplatin or paclitaxel in cervical cancer. (A) Schematic diagram shows a coculture system of M2 macrophage-like THP-1 cells and cancer cells. Cell viabilities of HeLa (B) and Siha (C) cells were analyzed by CCK-8. Cell invasion of HeLa (D) and Siha (E) cells was analyzed by Transwell assay. The images of Transwell assay are provided in Supplemental . *P < 0.05, **P < 0.01.

Article Snippet: Diluted primary CD200 rabbit antibody (#23451S, 1:200, Cell Signaling Technology, Shanghai, China), CD200R rabbit-antibody (1:400, #DF4715, Affinity, China), Ki-67 rabbit-antibody (1:400, #9027, Cell Signaling Technology) and CTSK rabbit antibody (1:400, #DF6614, Affinity, China) were incubated with the section at 4 °C for overnight.

Techniques: CCK-8 Assay, Transwell Assay

Journal: Heliyon

Article Title: CD200-CD200R affects cisplatin and paclitaxel sensitivity by regulating cathepsin K-mediated p65 NF-κB signaling in cervical cancer

doi: 10.1016/j.heliyon.2023.e19220

Figure Lengend Snippet: CD200-CD200R affects the chemosensitivity of cervical cancer cells through regulating the CTSK-mediated p65 NF-κB pathway. Cancer cells were treated with 0.1 μg/mL rCTSK as a control to compare the effects of C200-CD200R on resistance to cisplatin or paclitaxel. Cell viabilities of HeLa (A) and Siha (B) were analyzed by CCK-8. Expression of p-p65 NF-κB/p65 NF-κB protein in HeLa (C) and Siha (D) was analyzed by Western blotting. The full length of blots was provided in Supplemental materials. **P < 0.01.

Article Snippet: Diluted primary CD200 rabbit antibody (#23451S, 1:200, Cell Signaling Technology, Shanghai, China), CD200R rabbit-antibody (1:400, #DF4715, Affinity, China), Ki-67 rabbit-antibody (1:400, #9027, Cell Signaling Technology) and CTSK rabbit antibody (1:400, #DF6614, Affinity, China) were incubated with the section at 4 °C for overnight.

Techniques: Control, CCK-8 Assay, Expressing, Western Blot

Journal: Heliyon

Article Title: CD200-CD200R affects cisplatin and paclitaxel sensitivity by regulating cathepsin K-mediated p65 NF-κB signaling in cervical cancer

doi: 10.1016/j.heliyon.2023.e19220

Figure Lengend Snippet: CD200-C200R affects the paclitaxel sensitivity in mouse xenografts through regulating CTSK. Stably transfected plasmid or CD200 HeLa cells were used to establish xenografts. After injection for 7 days, 3.6 mg/kg MK-0822 (CTSK inhibitor) was orally treated every week, and 5 mg/kg paclitaxel was administrated by tail vein twice a week. (A) Tumor size was measured every week. (B) Images of xenografts and tumor weight. (C) Positive expression of Ki-67 in tumor tissues of each group. Scale = 50 μm. NS: no significance. **P < 0.01.

Article Snippet: Diluted primary CD200 rabbit antibody (#23451S, 1:200, Cell Signaling Technology, Shanghai, China), CD200R rabbit-antibody (1:400, #DF4715, Affinity, China), Ki-67 rabbit-antibody (1:400, #9027, Cell Signaling Technology) and CTSK rabbit antibody (1:400, #DF6614, Affinity, China) were incubated with the section at 4 °C for overnight.

Techniques: Stable Transfection, Transfection, Plasmid Preparation, Injection, Expressing

Journal: Heliyon

Article Title: CD200-CD200R affects cisplatin and paclitaxel sensitivity by regulating cathepsin K-mediated p65 NF-κB signaling in cervical cancer

doi: 10.1016/j.heliyon.2023.e19220

Figure Lengend Snippet: A CD200-CD200R mediated feedback loop between cancer cells and M2-tumor-associated macrophages (TAMs) in cervical cancer.

Article Snippet: Diluted primary CD200 rabbit antibody (#23451S, 1:200, Cell Signaling Technology, Shanghai, China), CD200R rabbit-antibody (1:400, #DF4715, Affinity, China), Ki-67 rabbit-antibody (1:400, #9027, Cell Signaling Technology) and CTSK rabbit antibody (1:400, #DF6614, Affinity, China) were incubated with the section at 4 °C for overnight.

Techniques:

Journal: Cells

Article Title: Maternal Immune Activation Sensitizes Male Offspring Rats to Lipopolysaccharide-Induced Microglial Deficits Involving the Dysfunction of CD200–CD200R and CX3CL1–CX3CR1 Systems

doi: 10.3390/cells9071676

Figure Lengend Snippet: The effect of MIA and the additional acute challenge with LPS on the gene expression of Cx3cl1 , Cx3cr1 , Cd200 and Cd200r in the frontal cortices ( A ) and hippocampi ( B ) of male Sprague-Dawley offspring at PND120. The mRNA levels were measured using qRT-PCR with n = 3–10 in each group. The results are presented as the average fold change ± SEM. * p < 0.05 vs. control + vehicle, # p < 0.05 vs. MIA responsive + vehicle, ^ p < 0.05 vs. MIA non-responsive + vehicle, & p < 0.05 vs. control + LPS.

Article Snippet: The protein levels of CX3CL1 (Cloud-Clone Corp., Katy, TX, USA), CX3CR1, CD200, CD200R, IL-6, and IL-4 (all from Cusabio, Houston, TX, USA) in the frontal cortices and hippocampi of the male rats were measured using commercially available ELISA kits.

Techniques: Gene Expression, Quantitative RT-PCR, Control

Journal: Cells

Article Title: Maternal Immune Activation Sensitizes Male Offspring Rats to Lipopolysaccharide-Induced Microglial Deficits Involving the Dysfunction of CD200–CD200R and CX3CL1–CX3CR1 Systems

doi: 10.3390/cells9071676

Figure Lengend Snippet: The effect of MIA and the additional acute challenge with LPS on the protein levels of CX3CL1, CX3CR1, CD200 and CD200R in the frontal cortices ( A ) and hippocampi ( B ) of male Sprague-Dawley offspring at PND120. n = 4–10 in each group. The results are presented as the means ± SEM. * p < 0.05 vs. control + vehicle, # p < 0.05 vs. MIA responsive + vehicle, ^ p < 0.05 vs. MIA non-responsive + vehicle, & p < 0.05 vs. control + LPS.

Article Snippet: The protein levels of CX3CL1 (Cloud-Clone Corp., Katy, TX, USA), CX3CR1, CD200, CD200R, IL-6, and IL-4 (all from Cusabio, Houston, TX, USA) in the frontal cortices and hippocampi of the male rats were measured using commercially available ELISA kits.

Techniques: Control

Journal: American journal of clinical pathology

Article Title: Diagnostic Utility of CD200 Immunohistochemistry in Distinguishing EBV-Positive Large B-Cell Lymphoma From Classic Hodgkin Lymphoma.

doi: 10.1093/ajcp/aqad053

Figure Lengend Snippet: FIGURE 2 Representative CD200 staining in Hodgkin and large B-cell lymphoma (LBCL). Representative images of H&E-stained sections and immunohistochemical stains for cases in each diagnostic category. All images ×400. CHL, classic Hodgkin lymphoma; DLBCL, diffuse large B-cell lymphoma; EBV, Epstein-Barr virus; NOS, not otherwise specified.

Article Snippet: Staining was performed on a Leica Bond-Max (Leica Biosystems) following antigen retrieval (EDTA pH 9.0, 20 minutes) with a goat anti-human CD200 polyclonal antibody (AF2724; R&D Systems) at a 1:150 dilution.

Techniques: Staining, Immunohistochemical staining, Diagnostic Assay, Virus

Journal: American journal of clinical pathology

Article Title: Diagnostic Utility of CD200 Immunohistochemistry in Distinguishing EBV-Positive Large B-Cell Lymphoma From Classic Hodgkin Lymphoma.

doi: 10.1093/ajcp/aqad053

Figure Lengend Snippet: FIGURE 3 CD200 can help differentiate Epstein-Barr virus−positive (EBV+) classic Hodgkin lymphoma (CHL) with frequent Reed-Sternberg (RS) cells and EBV+ large B-cell lymphoma (LBCL). Representative images of H&E-stained sections and immunohistochemical stains for a case of EBV+ (CHL) with frequent RS cells and a case of EBV+ LBCL. In both cases, lesional cells were positive for CD30 and EBER and showed variable positivity for CD20. However, CD200 was positive in the case of EBV+ CHL and negative in the case of EBV+ LBCL. All images ×400.

Article Snippet: Staining was performed on a Leica Bond-Max (Leica Biosystems) following antigen retrieval (EDTA pH 9.0, 20 minutes) with a goat anti-human CD200 polyclonal antibody (AF2724; R&D Systems) at a 1:150 dilution.

Techniques: Virus, Staining, Immunohistochemical staining

Journal: American journal of clinical pathology

Article Title: Diagnostic Utility of CD200 Immunohistochemistry in Distinguishing EBV-Positive Large B-Cell Lymphoma From Classic Hodgkin Lymphoma.

doi: 10.1093/ajcp/aqad053

Figure Lengend Snippet: FIGURE 4 CD200 can help differentiate Epstein-Barr virus−positive (EBV+) large B-cell lymphoma (LBCL) with rare neoplastic cells from EBV+ classic Hodgkin lymphoma (CHL). Representative images of H&E-stained sections and immunohistochemical stains for a case of EBV+ CHL and a case of EBV+ LBCL with rare neoplastic cells and CHL-like morphology. In both cases, lesional cells were weakly positive for CD20 and CD30. However, CD200 was positive in the case of EBV+ CHL and negative in the case of EBV+ LBCL (arrows). All images ×600.

Article Snippet: Staining was performed on a Leica Bond-Max (Leica Biosystems) following antigen retrieval (EDTA pH 9.0, 20 minutes) with a goat anti-human CD200 polyclonal antibody (AF2724; R&D Systems) at a 1:150 dilution.

Techniques: Virus, Staining, Immunohistochemical staining

Journal: American journal of clinical pathology

Article Title: Diagnostic Utility of CD200 Immunohistochemistry in Distinguishing EBV-Positive Large B-Cell Lymphoma From Classic Hodgkin Lymphoma.

doi: 10.1093/ajcp/aqad053

Figure Lengend Snippet: FIGURE 5 A single case of CD200-negative classic Hodgkin lymphoma (CHL). The single case of CHL in which Reed-Sternberg cells were negative for CD200 was Epstein-Barr virus positive (EBV+), HIV associated, and refractory to CHL-directed therapy, raising the possibility that this may in fact have represented EBV+ large B-cell lymphoma. Arrows highlight large abnormal cells that show weak positivity for PAX5 (B) and variable positivity for CD20 (C) and are negative for CD200 (F). A, H&E. D, CD30. E, EBER. All images ×400.

Article Snippet: Staining was performed on a Leica Bond-Max (Leica Biosystems) following antigen retrieval (EDTA pH 9.0, 20 minutes) with a goat anti-human CD200 polyclonal antibody (AF2724; R&D Systems) at a 1:150 dilution.

Techniques: Virus

Journal: Aging cell

Article Title: Hedgehog signaling maintains hair follicle stem cell phenotype in young and aged human skin.

doi: 10.1111/j.1474-9726.2009.00526.x

Figure Lengend Snippet: Fig. 1 Characterization of human bulge cell markers. (A) CD200+ cells were stained in red by immunohistochemistry (magnification = 100·). Black rectangle identifies boundaries of 400· magnification shown in left inset. Positive cells are located in the outer root sheath of the hair follicle (HF), between insertions of sebaceous glands (SG) and arrector pili muscle (AP). Asterisk shows lumen of the HF (hair shaft was lost during staining). (B) Consecutive sections of human scalp skin biopsies from young individuals were stained for KRT15, CD200, and KRT19 by immunohistochemistry. Upper and lower limits of CD200 positive staining superimpose with those of KRT19 and KRT15 respectively (highlighted by black lines). Daggers indicate hair shafts remnants. (C) Schematic representation of localization of protein used as human bulge cell markers. Bulge cells are CD200+ ⁄ KRT15+ ⁄ KRT19+.

Article Snippet: Total HF cell suspensions were incubated with human IgGs (Sigma, St Louis, MO, USA) for 15 min at 4 C, and labeled with a mouse anti-CD200 antibody (AbD Serotec, Raleigh, NC, USA) at 4 C for 15 min. After three washes with MACS buffer, cells were incubated with microbeads coupled with rat antimouse IgG1 antibodies (Miltenyi Biotec) at 4 C for 15 min. After three washes with MACS buffer, cell suspension was loaded onto a column containing a ferromagnetic matrix exposed to the magnetic field of a permanent magnet.

Techniques: Staining, Immunohistochemistry

Journal: Aging cell

Article Title: Hedgehog signaling maintains hair follicle stem cell phenotype in young and aged human skin.

doi: 10.1111/j.1474-9726.2009.00526.x

Figure Lengend Snippet: Fig. 2 Characterization of isolated human bulge cells from young human skin. (A) CD200, KRT15 and KRT19 mRNA levels were measured by Q-PCR on CD200+ (bulge) and CD200) (differentiated) cells of the hair follicle (HF) in young (< 40 years) individuals. N = 6; *P < 0.05. (B) Transglutaminase 1 (TGM1) mRNA levels in same cell populations. N = 3; P = 0.022.

Article Snippet: Total HF cell suspensions were incubated with human IgGs (Sigma, St Louis, MO, USA) for 15 min at 4 C, and labeled with a mouse anti-CD200 antibody (AbD Serotec, Raleigh, NC, USA) at 4 C for 15 min. After three washes with MACS buffer, cells were incubated with microbeads coupled with rat antimouse IgG1 antibodies (Miltenyi Biotec) at 4 C for 15 min. After three washes with MACS buffer, cell suspension was loaded onto a column containing a ferromagnetic matrix exposed to the magnetic field of a permanent magnet.

Techniques: Isolation

Journal: Aging cell

Article Title: Hedgehog signaling maintains hair follicle stem cell phenotype in young and aged human skin.

doi: 10.1111/j.1474-9726.2009.00526.x

Figure Lengend Snippet: Fig. 3 Additional markers of young human bulge keratinocytes. (A, B) Frozen scalp skin sections from young individuals were immunostained for CD34 (A) and follistatin (B). Original magnification = 100·. Black rectangle indicates location of left inset (magnification 400·). Black arrows represent upper and lower limits of bulge area. Representative of at least four individuals. (C) Quantitative expression of gene transcripts for various potential hair follicle (HF) stem cell markers in bulge (CD200+) and differentiated (CD200)) cells of the HF in young human skin. N = 6; *P < 0.05; NS = not significant vs. bulge cells. Gas1, EGFR, CCN2 and S100A4 are represented in separate bar graphs to account for higher basal expression levels.

Article Snippet: Total HF cell suspensions were incubated with human IgGs (Sigma, St Louis, MO, USA) for 15 min at 4 C, and labeled with a mouse anti-CD200 antibody (AbD Serotec, Raleigh, NC, USA) at 4 C for 15 min. After three washes with MACS buffer, cells were incubated with microbeads coupled with rat antimouse IgG1 antibodies (Miltenyi Biotec) at 4 C for 15 min. After three washes with MACS buffer, cell suspension was loaded onto a column containing a ferromagnetic matrix exposed to the magnetic field of a permanent magnet.

Techniques: Expressing

Journal: Aging cell

Article Title: Hedgehog signaling maintains hair follicle stem cell phenotype in young and aged human skin.

doi: 10.1111/j.1474-9726.2009.00526.x

Figure Lengend Snippet: Fig. 5 Aging does not alter bulge marker expression in human skin. Total RNA from bulge (CD200+) and differentiated (CD200)) cells of the hair follicle (HF) were isolated from scalp skin biopsies of young (< 40 years) and aged (> 70 years) individuals, and analyzed by Q-PCR for human HF stem cell markers. (A) Quantitative expression of CD200, KRT15 and KRT19 human bulge cell markers. (B) Direct comparison of gene transcript levels of various potential additional bulge cell markers between young and aged individuals. Gas1, EGFR and CCN2 are represented in separate bar graphs to account for higher expression levels. N = 6; *P<0.05; NS = not significant vs. young counterpart.

Article Snippet: Total HF cell suspensions were incubated with human IgGs (Sigma, St Louis, MO, USA) for 15 min at 4 C, and labeled with a mouse anti-CD200 antibody (AbD Serotec, Raleigh, NC, USA) at 4 C for 15 min. After three washes with MACS buffer, cells were incubated with microbeads coupled with rat antimouse IgG1 antibodies (Miltenyi Biotec) at 4 C for 15 min. After three washes with MACS buffer, cell suspension was loaded onto a column containing a ferromagnetic matrix exposed to the magnetic field of a permanent magnet.

Techniques: Marker, Expressing, Isolation, Comparison

Journal: Aging cell

Article Title: Hedgehog signaling maintains hair follicle stem cell phenotype in young and aged human skin.

doi: 10.1111/j.1474-9726.2009.00526.x

Figure Lengend Snippet: Fig. 6 Hedgehog (Hh) pathway components are selectively expressed in human bulge cells vs. differentiated hair follicle (HF) keratinocytes (KCs). Transcript levels of Hh pathway components and target genes were quantified by Q-PCR in bulge (CD200+) and differentiated (CD200)) cells of human HFs. N = 6; (A) young and (B) aged individuals. *P < 0.05; NS = not significant vs. differentiated HF KCs. (C) Quantitative expression of levels of Hh pathway components and target genes transcripts in bulge cells of young and aged individuals. NS = not significant vs. young counterpart.

Article Snippet: Total HF cell suspensions were incubated with human IgGs (Sigma, St Louis, MO, USA) for 15 min at 4 C, and labeled with a mouse anti-CD200 antibody (AbD Serotec, Raleigh, NC, USA) at 4 C for 15 min. After three washes with MACS buffer, cells were incubated with microbeads coupled with rat antimouse IgG1 antibodies (Miltenyi Biotec) at 4 C for 15 min. After three washes with MACS buffer, cell suspension was loaded onto a column containing a ferromagnetic matrix exposed to the magnetic field of a permanent magnet.

Techniques: Expressing

Journal: Aging cell

Article Title: Hedgehog signaling maintains hair follicle stem cell phenotype in young and aged human skin.

doi: 10.1111/j.1474-9726.2009.00526.x

Figure Lengend Snippet: Fig. 7 The hedgehog transmembrane receptor and target gene product Ptch is expressed in CD200+ cells of the bulge area of human hair follicles. (A, B) Ptch protein expression was stained by immunohistochemistry 100· magnification (A). Black rectangle in A identifies boundaries of 200· magnification shown in B. (C) Double immunofluorescence staining of human bulge area. Human scalp sections were stained with CD200 (green) and Ptch (red). Overlay shows that Ptch is expressed by CD200+ cells of the human bulge area. Magnification = 400·. Representative of at least three individuals.

Article Snippet: Total HF cell suspensions were incubated with human IgGs (Sigma, St Louis, MO, USA) for 15 min at 4 C, and labeled with a mouse anti-CD200 antibody (AbD Serotec, Raleigh, NC, USA) at 4 C for 15 min. After three washes with MACS buffer, cells were incubated with microbeads coupled with rat antimouse IgG1 antibodies (Miltenyi Biotec) at 4 C for 15 min. After three washes with MACS buffer, cell suspension was loaded onto a column containing a ferromagnetic matrix exposed to the magnetic field of a permanent magnet.

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Pharmaceutics

Article Title: Minoxidil Nanoparticles Targeting Hair Follicles Enhance Hair Growth in C57BL/6 Mice

doi: 10.3390/pharmaceutics14050947

Figure Lengend Snippet: Microscopic effects of MXD formulations on skin tissue (hair bulge and hair bulb) of C57BL/6 mice at 8 d after the start of repetitive applications (once a day). ( A ) Images of H&E-stained skin tissue specimens from C57BL/6 mice (bars indicate 100 µm). ( B ) High magnification microscope images in the areas designated by the squares in Figure A (bars indicate 50 µm). ( C ) Optical images of immunostaining for CD200 in the serial segments shown in Figure A (bars indicate 100 µm). ( D ) High magnification microscope images of the areas designated by the squares in Figure C (bars indicate 50 µm). Vehicle, only the vehicle used to prepare the MXD-MPs and MXD-NPs formulations was applied. MXD-MPs, MXD-MPs-applied mice. MXD-NPs, MXD-NPs-applied mice. CA-MXD, CA-MXD-applied mice. The arrow shows the expression of CD200. CD200 is stained brown; the expression of CD200 was enhanced in the MXD-NPs group, compared with the CA-MXD group.

Article Snippet: Paraffin sections were obtained from the fixed mouse back skin tissues in the usual manner and the product certificate and were incubated with anti-CD200 rabbit polyclonal antibody (1:100; Cat No. 14057-1-AP; Proteintech, Rosemont, IL, USA) for 1 h at 37 °C.

Techniques: Staining, Microscopy, Immunostaining, Expressing

Journal: International Journal of Nanomedicine

Article Title: Preparation, characterization, and transfection efficiency of low molecular weight polyethylenimine-based nanoparticles for delivery of the plasmid encoding CD200 gene

doi: 10.2147/ijn.s140734

Figure Lengend Snippet: Figure 6 Cellular toxicity and transfection efficiency. Notes: Viability and gene transfer ability of unmodified and modified PEI derivatives complexed with pDNA encoding CD200 gene at polymer:plasmid (weight:weight) ratios of 0.25, 4, and 8 determined in triplicate in SH-SY5Y cell cultures in 96-well plates. (A) Cell survival was assayed by the MTT method, and expressed as the percentages of cell viability. (B) CD200 expression level measured by flow cytometry. *P,0.05, PEI derivative compared to unmodified parent polymer at the same C/P ratio. (n=3; error bars represent mean ± standard deviation). Abbreviations: C/P, carrier to plasmid ratio; pDNA, plasmid DNA; PEI, polyethylenimine; PEI-SUC, PEI-succinate conjugate; PEI-SUC-PEI, PEI-succinate-PEI conjugate.

Article Snippet: Human CD200 plasmid (pCMV-XL5-hCD200) was obtained from OriGene (Rockville, MD, USA).

Techniques: Transfection, Modification, Polymer, Plasmid Preparation, Expressing, Flow Cytometry, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Preparation, characterization, and transfection efficiency of low molecular weight polyethylenimine-based nanoparticles for delivery of the plasmid encoding CD200 gene

doi: 10.2147/ijn.s140734

Figure Lengend Snippet: Figure 7 In vivo imaging of MS induced mice using fluorescently labeled plasmid encoding CD200 at C/P ratio 8. Notes: Mice were injected with the final volume of 100 μL of the polyplex formulation. (A) The control mice received 100 μL of HBG buffer. (B–I) Mice were injected with the polyplexes and imaging was performed at the time points of 0, 15, 30, 60, 120, 180 min and 24 h post injection. Magnification ×10. Abbreviations: C/P, carrier to plasmid ratio; MS, multiple sclerosis; HBG, HEPES buffered glucose.

Article Snippet: Human CD200 plasmid (pCMV-XL5-hCD200) was obtained from OriGene (Rockville, MD, USA).

Techniques: In Vivo Imaging, Labeling, Plasmid Preparation, Injection, Formulation, Control, Imaging

Journal: Neoplasia (New York, N.Y.)

Article Title: A Truncated form of CD200 (CD200S) Expressed on Glioma Cells Prolonged Survival in a Rat Glioma Model by Induction of a Dendritic Cell-Like Phenotype in Tumor-Associated Macrophages.

doi: 10.1016/j.neo.2016.02.006

Figure Lengend Snippet: Figure 3. Characterization of established cell lines expressing CD200L and CD200S. (A–C) Immunocytochemical confirmation of CD200 expression with antibodies recognizing N- (OX2) or C-termini of CD200. The C6 cell line transfected with an empty vector (A, C6-e) did not display positive immunostaining against either antibody. (B) The C6-L cell line was positive for both antibodies. (C) The C6-S line was only reactive to the C-terminal antibody. (D) qPCR confirmed the specific expression of CD200L or CD200S. A primer set amplifying cDNA derived from both CD200L- and CD200S-mRNA was called common ‘Com’ (see Supplementary Table 1). The other primer set ‘L’ amplified cDNA only derived from CD200L-mRNA. C6-e cells expressed none of the cDNA, C6-L cells expressed both, and C6-S cells expressed cDNA amplified only by the Com primer set. (E) Growth curves of each cell line. Data from four independent cultures are expressed as means ± SD except for the data of Empty at 5 days, which is expressed as mean −SD.

Article Snippet: RNA splicing generates CD200S with a shorter C–C′ loop or CDR2, all of which are responsible for the binding to a cells to generate C6-L or C6-S cell lines. fragments of CD200L or CD200S were amplified by PCR using CD200 cDNA purchased from Origene (Rockville, MD) with the following cloning primers containing BamHI and NotI respectively (see Figure 1): CD200L; forward 5′-GGA TCC ATG GGC AGT CCG GTA TTC AG-3′ and reverse 5′-ATG CGG CCG CTT ATT TCA TTC TTT GCA TCC CCT G-3′, CD200S; forward 5′-ACG GAT CCA TGA TCA CTT ACA GCA AAG CC-3′ and the same reverse primer as for CD200L.

Techniques: Expressing, Transfection, Plasmid Preparation, Immunostaining, Derivative Assay, Amplification

Journal: Neoplasia (New York, N.Y.)

Article Title: A Truncated form of CD200 (CD200S) Expressed on Glioma Cells Prolonged Survival in a Rat Glioma Model by Induction of a Dendritic Cell-Like Phenotype in Tumor-Associated Macrophages.

doi: 10.1016/j.neo.2016.02.006

Figure Lengend Snippet: Figure 7. Expression levels in tumors and isolated TAMs of mRNA encoding molecules related to CD200, DCs, lymphocytes and T cells, as revealed by qPCR. Data are shown in an alphabetical order. (A) cDNA was prepared from tumor masses dissected out 21 days after transplantation. CD200-mRNA amplified with ‘CD200 com’ primers was highly expressed by C6-L and C6-S tumors, whereas the CD200L-mRNA level was high only in the C6-L-tumors. The C6-S-tumors expressed DC markers, MHC class II α, CD11c, and CD80 at significantly higher levels than other tumors. Expression of mRNA encoding lymphocyte markers CD8α and β, CTL-related apoptosis-inducing molecules, granzyme, and perforin was significantly higher in the C6-S tumors. Despite not showing statistical significance, CD4 and IFNγ-mRNA were also highly expressed in the C6-S tumors. CLEC9A, which is expressed by myeloid DCs and responsible for cross-presentation, and chemokines (CCL12, CXCL10 and CXCL16), which are invovlved in recruitment of monocytes and lymphocytes, are also highly expressed in the C6-S tumor. (B) Shortly after isolation from the tumor mass, TAMs were subjected to qPCR analysis. TAMs from the C6-S-tumors expressed MHC class II α-, and CD11c-mRNA was expressed at significantly higher levels than in TAMs from other tumors. CD86-mRNA expression in TAMs from the C6-S tumors was also high. Data from 3 tumors of each cell lines are expressed as means ± SEM. *P b .05, **P b .01, ***P b .001 versus CD200S; #P b .05, ##P b .01 versus CD200L.

Article Snippet: RNA splicing generates CD200S with a shorter C–C′ loop or CDR2, all of which are responsible for the binding to a cells to generate C6-L or C6-S cell lines. fragments of CD200L or CD200S were amplified by PCR using CD200 cDNA purchased from Origene (Rockville, MD) with the following cloning primers containing BamHI and NotI respectively (see Figure 1): CD200L; forward 5′-GGA TCC ATG GGC AGT CCG GTA TTC AG-3′ and reverse 5′-ATG CGG CCG CTT ATT TCA TTC TTT GCA TCC CCT G-3′, CD200S; forward 5′-ACG GAT CCA TGA TCA CTT ACA GCA AAG CC-3′ and the same reverse primer as for CD200L.

Techniques: Expressing, Isolation, Transplantation Assay, Amplification