Journal: Journal of translational medicine

Article Title: The clinical importance of tumour-infiltrating macrophages and dendritic cells in periampullary adenocarcinoma differs by morphological subtype.

doi: 10.1186/s12967-017-1256-y

Figure Lengend Snippet: Fig. 1 Sample immunohistochemical images. Sample images of immunohistochemical staining of CD1a+ TIDC in a PB-type, b I-type tumour, CD68+ TAM in c PB-type, d I-type tumour, CD163+ TAM in e PB-type, f I-type tumour, and MARCO+ TAM in g PB-type, h I-type. Scale bar represents 20 μm

Article Snippet: For immunohistochemical (IHC) analysis of CD1a, CD68, CD163 and MARCO, 4 μm TMA-sections were pretreated using ULTRA Cell Conditioning Solution 1, pH 8.5 (Ventana Medical Systems Inc., Tucson, AZ, USA) for heat induced epitope retrieval, and stained in a Ventana BenchMark stainer (Ventana Medical Systems Inc.) with the following antibodies: CD1a: clone NCL-CD1a-220, diluted 1:25, LEICA Biosystems, Newcastle, UK, CD68: clone KP1, diluted 1:1000, Dako, Glostrup, Denmark, CD163: clone 10D6 diluted 1:200 Novus Biologicals, Abingdon, United Kingdom, MARCO clone HPA063793, diluted 1:250, Atlas Antibodies, Bromma, Sweden.

Techniques: Immunohistochemical staining, Staining

Journal: Journal of translational medicine

Article Title: The clinical importance of tumour-infiltrating macrophages and dendritic cells in periampullary adenocarcinoma differs by morphological subtype.

doi: 10.1186/s12967-017-1256-y

Figure Lengend Snippet: Fig. 2 Kaplan–Meier estimates of survival according to CD1a+ TIDC density. Kaplan–Meier estimates of 5-year overall survival according to high and low TIDCs density in a the entire cohort, b in I-type tumours and c in PB-type tumours

Article Snippet: For immunohistochemical (IHC) analysis of CD1a, CD68, CD163 and MARCO, 4 μm TMA-sections were pretreated using ULTRA Cell Conditioning Solution 1, pH 8.5 (Ventana Medical Systems Inc., Tucson, AZ, USA) for heat induced epitope retrieval, and stained in a Ventana BenchMark stainer (Ventana Medical Systems Inc.) with the following antibodies: CD1a: clone NCL-CD1a-220, diluted 1:25, LEICA Biosystems, Newcastle, UK, CD68: clone KP1, diluted 1:1000, Dako, Glostrup, Denmark, CD163: clone 10D6 diluted 1:200 Novus Biologicals, Abingdon, United Kingdom, MARCO clone HPA063793, diluted 1:250, Atlas Antibodies, Bromma, Sweden.

Techniques:

Journal: Inflammatory Bowel Diseases

Article Title: CD1a-Expressing Monocytes as Mediators of Inflammation in Ulcerative Colitis

doi: 10.1093/ibd/izy073

Figure Lengend Snippet: Incubation of PBMCs with PC affects frequencies of monocytes, macrophages, and CD4 T cells. A, Flow cytometric analysis of PBMCs from UC patients (n = 13) and non-UC donors (n = 28) that were incubated in the absence (control n = 39) or presence of PC (n = 41) and PHA (n = 40) for 48 hours depicted as boxplot diagrams. B, Cell trace CFSE dilution of CD14+ CD1a+ monocytes. C, Correlation analysis of frequencies of CD14+ CD1a+ monocytes and Th1 of Th2 cells depicted as scatter plots. The method was Pearson; numbers display correlation coefficients (cor-values) and P values. D, Flow cytometric analysis of CD4+ T cells for intracellular expression of IFNγ, IL-4, or IL-17 depicted as boxplots. PBMCs (n = 4) were incubated in the absence or presence of PC and anti-CD1a antibody. For comparison of groups, ANOVA followed by Tukey’s HSD was conducted. Boxes represent upper and lower quartiles, whiskers represent variability, and outliers are plotted as individual points ( *** 0.001; ** 0.01; *0.05).

Article Snippet: To analyze the effect of an anti-CD1a antibody on cells stimulated with PC, cells were incubated for 1 hour with 10 μg/mL of anti-human CD1a (Bio X Cell, West Lebanon, USA) and 10 μg/mL of a corresponding isotype antibody (Biolegend, San Diego, CA, USA) before 200 μg/mL of PC (Sigma-Aldrich, St. Louis, USA) was added.

Techniques: Incubation, Control, Expressing, Comparison

Journal: Inflammatory Bowel Diseases

Article Title: CD1a-Expressing Monocytes as Mediators of Inflammation in Ulcerative Colitis

doi: 10.1093/ibd/izy073

Figure Lengend Snippet: Anti-CD1a antibodies affect frequencies of CD4+ T cells. Flow cytometric analysis of PBMCs from 2 different donors that were incubated in triplicate for 5 days with PC in the presence (n = 6) or absence of anti-CD1a antibodies (n = 6) or PHA (n = 6) depicted as boxplot diagrams. For comparison of groups, ANOVA followed by Tukey’s HSD was conducted. Boxes represent upper and lower quartiles, whiskers represent variability, and outliers are plotted as individual points ( *** 0.001; ** 0.01; *0.05).

Article Snippet: To analyze the effect of an anti-CD1a antibody on cells stimulated with PC, cells were incubated for 1 hour with 10 μg/mL of anti-human CD1a (Bio X Cell, West Lebanon, USA) and 10 μg/mL of a corresponding isotype antibody (Biolegend, San Diego, CA, USA) before 200 μg/mL of PC (Sigma-Aldrich, St. Louis, USA) was added.

Techniques: Incubation, Comparison

Journal: Inflammatory Bowel Diseases

Article Title: CD1a-Expressing Monocytes as Mediators of Inflammation in Ulcerative Colitis

doi: 10.1093/ibd/izy073

Figure Lengend Snippet: Ex vivo analysis of CD14+ CD1a+ monocytes for co-expression of CCR2, CD86, and TSLPR. Flow cytometric analysis of PBMCs from UC patients (n = 21) and non-UC (n = 9) donors depicted as boxplot diagrams. Boxes represent upper and lower quartiles, whiskers represent variability, and outliers are plotted as individual points. For comparison of groups, a Student t test was performed ( *** 0.001; ** 0.01; *0.05).

Article Snippet: To analyze the effect of an anti-CD1a antibody on cells stimulated with PC, cells were incubated for 1 hour with 10 μg/mL of anti-human CD1a (Bio X Cell, West Lebanon, USA) and 10 μg/mL of a corresponding isotype antibody (Biolegend, San Diego, CA, USA) before 200 μg/mL of PC (Sigma-Aldrich, St. Louis, USA) was added.

Techniques: Ex Vivo, Expressing, Comparison

Journal: Inflammatory Bowel Diseases

Article Title: CD1a-Expressing Monocytes as Mediators of Inflammation in Ulcerative Colitis

doi: 10.1093/ibd/izy073

Figure Lengend Snippet: TAG and cholesterol levels increase in the NSG-UC mouse model and correlate with CD1a-expressing monocytes. NSG-UC mice were engrafted with PBMCs derived from UC patients (n = 4), challenged with 10% ethanol at day 8 and 50% ethanol at days 15 and 18. A, TAG and cholesterol levels depicted as boxplot diagrams; nchallenged group (control, n = 12), ethanol challenged group (ethanol, n = 21). Whiskers represent variability, and outliers are plotted as individual points. B, Correlation analysis of CD14+ CD1a+ monocytes with TAG and cholesterol levels depicted as scatter plots. (TAG n = 24, cholesterol n = 25). The method was Spearman; numbers display correlation coefficients ( rho values) and P values.

Article Snippet: To analyze the effect of an anti-CD1a antibody on cells stimulated with PC, cells were incubated for 1 hour with 10 μg/mL of anti-human CD1a (Bio X Cell, West Lebanon, USA) and 10 μg/mL of a corresponding isotype antibody (Biolegend, San Diego, CA, USA) before 200 μg/mL of PC (Sigma-Aldrich, St. Louis, USA) was added.

Techniques: Expressing, Derivative Assay, Control

Journal: Inflammatory Bowel Diseases

Article Title: CD1a-Expressing Monocytes as Mediators of Inflammation in Ulcerative Colitis

doi: 10.1093/ibd/izy073

Figure Lengend Snippet: NSG-UC mice benefit from treatment with anti-CD1a antibodies. NSG mice were engrafted with PBMCs derived from UC patients (NSG-UC, n = 3) and 1 non-UC donor (NSG-non-UC). Mice were challenged with 10% ethanol at day 8 and 50% ethanol at days 15 and 18 and treated with 30 µg of anti-CD1a antibody or isotype control at days 7, 14, and 17. Control: NSG-non-UC, n = 4; NSG-UC, n = 8. Challenged control (ethanol + isotype): NSG-non-UC, n = 4; NSG-UC, n = 16. Challenged and treated with anti-CD1a antibody (ethanol + anti-CD1a): NSG-non-UC, n = 4; NSG-UC, n = 16. A, Clinical colon and histological scores of NSG-UC mice depicted as boxplot diagrams. B, Macrophotographs of colons at autopsy. a, NSG-UC unchallenged control. b, NSG-non-UC challenged control. c, NSG-UC challenged control, NSG-UC. d, Challenged and treated with anti-CD1a antibody. C, Photomicrographs of H&E-stained sections of distal parts of the colon from mice. Arrows indicate edema and influx of inflammatory cells. D, Frequencies of human leucocytes isolated from colons of mice. Samples from each group were pooled. Mean values are given; error bars are SD. Quantification was performed using flow cytometric analysis. Sample sizes: control n = 3, ethanol + isotype n = 3, ethanol + anti-CD1a n = 3. E, mRNA expression levels of mTARC, mTGFß1, HGF, and hIFNγ depicted as boxplots. Lg: delta CT, logarithmic delta cycle threshold. Boxes represent upper and lower quartiles. Whiskers represent variability, and outliers are plotted as individual points. Labels given on x-axes on the bottom row apply to all charts. For comparison of groups, ANOVA followed by Tukey’s HSD was conducted.

Article Snippet: To analyze the effect of an anti-CD1a antibody on cells stimulated with PC, cells were incubated for 1 hour with 10 μg/mL of anti-human CD1a (Bio X Cell, West Lebanon, USA) and 10 μg/mL of a corresponding isotype antibody (Biolegend, San Diego, CA, USA) before 200 μg/mL of PC (Sigma-Aldrich, St. Louis, USA) was added.

Techniques: Derivative Assay, Control, Staining, Isolation, Expressing, Comparison

Journal: Inflammatory Bowel Diseases

Article Title: CD1a-Expressing Monocytes as Mediators of Inflammation in Ulcerative Colitis

doi: 10.1093/ibd/izy073

Figure Lengend Snippet: Treatment of NSG-UC mice with anti-CD1a antibodies affects CD14+ CD1a+ monocytes. NSG mice were engrafted and treated as in . Human leucocytes were isolated from mouse spleen and subjected to flow cytometric analysis. A, Frequencies of CD14+ CD1a+ and CD11b+ CD1a+ cells depicted as boxplot diagrams. Boxes represent upper and lower quartiles. Whiskers represent variability, and outliers are plotted as individual points. B, Correlation analysis of frequencies of CD14+ CD1a+ monocytes with subsets of B and T cells cells depicted as scatter plots. The method was Spearman; numbers display correlation coefficients ( rho values) and P values.

Article Snippet: To analyze the effect of an anti-CD1a antibody on cells stimulated with PC, cells were incubated for 1 hour with 10 μg/mL of anti-human CD1a (Bio X Cell, West Lebanon, USA) and 10 μg/mL of a corresponding isotype antibody (Biolegend, San Diego, CA, USA) before 200 μg/mL of PC (Sigma-Aldrich, St. Louis, USA) was added.

Techniques: Isolation