Journal: Veterinary Research

Article Title: Interactions between natural killer cells and dendritic cells favour T helper1-type responses to BCG in calves

doi: 10.1186/s13567-016-0367-4

Figure Lengend Snippet: BCG is taken up by bovine monocyte-derived DCs resulting in increased the expression of MHC class II, CD40 and CD80 and production of the Th1 polarising cytokine IL-12. Monocyte-derived DCs were cultured for 3 days and infected with FITC-labelled BCG (MOI 5) for 42 h. FACS plots from one representative animal ( A ) illustrate the percentage uptake of BCG-FITC by uninfected DCs (blue histogram) and BCG-infected DCs (pink histogram). Gates were set using uninfected DCs. Pooled data from four calves ( A ) illustrate the average percentage uptake of BCG-FITC ± SD by uninfected DCs (lighter bar) and BCG-infected DCs (darker bar). Uninfected and BCG-infected DCs were labelled with mAbs for MHC class II, CD40 and CD80 and analysed by flow cytometry. FACS plots from one representative animal show the expression of MHC class II ( B ), CD40 ( C ) and CD80 ( D ) by uninfected (blue histograms) and BCG-infected DCs (pink histograms). Positive cells were identified based on FMO controls (green histograms). Pooled data from four calves indicates the average MFI ± SD of MHC class II ( B ), CD40 ( C ) and CD80 ( D ) expression by uninfected (lighter bars) and BCG-infected DCs (darker bars). Supernatants were retrieved from uninfected and BCG-infected DCs and assayed for IL-12 production by ELISA. Pooled data from five calves represent the average levels ± SD of IL-12 BU/mL produced by uninfected (lighter bars) and BCG-infected DCs (darker bars) ( E ). Data were normally distributed ( p > 0.05) and significance between uninfected and BCG-infected DCs was assessed using a 2-sample t test. p < 0.05*, p < 0.001***.

Article Snippet: Mouse anti-bovine MHC class II-PE (CC158, IgG2a, Bio-Rad AbD Serotec), mouse anti-bovine CD40-FITC (IL-A156, IgG1, Bio-Rad AbD Serotec) and mouse anti-bovine CD80 (IL-A159, Bio-Rad AbD Serotec) indirectly conjugated to goat anti-mouse IgG1-AF647 (Life Technologies, UK) were used to determine DC expression of MHC class II, CD40 and CD80 respectively.

Techniques: Derivative Assay, Expressing, Cell Culture, Infection, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Produced

Journal: Frontiers in Immunology

Article Title: Regulatory T Cells Diminish HIV Infection in Dendritic Cells – Conventional CD4 + T Cell Clusters

doi: 10.3389/fimmu.2014.00199

Figure Lengend Snippet: DC and Tcon phenotype .

Article Snippet: Cells in co-cultures were harvested, treated with 20 μg/ml of human IgG to block Fc receptors, and stained with anti-HLA-DR eFluor450 (Clone L243; eBiosciences, San Diego, CA, USA), anti-CD80 AF-647 (Clone MEM-233, AbD Serotec, Raleigh, NC, USA), anti-CD83 PE-Cy7 (Clone HB15e, eBiosciences), anti-CD86 AF-647 [Clone 2331 (FUN-1), BD; San Diego, CA, USA], anti CD40-APC-H7 (Clone 5C3, BD), anti-CD25 AF700 (Clone M-A251, BD), anti-CD69 PE-Cy7 (Clone FN50, Biolegend, San Diego, CA, USA), anti-CD134 PerCP-Cy5.5 (Clone ACT35, Biolegend), and Live/Dead ® Fixable Aqua Dead Cell Stain Kit (Invitrogen) for 30 min at room temperature in PBS containing 2% fetal calf-serum and 0.1% sodium azide.

Techniques:

Journal: Frontiers in Immunology

Article Title: Regulatory T Cells Diminish HIV Infection in Dendritic Cells – Conventional CD4 + T Cell Clusters

doi: 10.3389/fimmu.2014.00199

Figure Lengend Snippet: Decreased actin polymerization and DC maturation by Treg is the effect most associated with their control of HIV replication . Comparison of Treg effect and combined CTLA-4/cAMP blockage on HIV replication, actin polymerization, DC maturation, and Tcon activation. (A) Blockade of Treg function on HIV infection or actin polymerization. (B) Blockade of Treg function on HIV infection or CD83 and CD40 expression. (C) Blockade of Treg function on HIV infection or CD25 and CD134 expression. Median of 6 (for actin polymerization) and 7 (for CD83, CD40, CD25, and CD134) experiment is shown.

Article Snippet: Cells in co-cultures were harvested, treated with 20 μg/ml of human IgG to block Fc receptors, and stained with anti-HLA-DR eFluor450 (Clone L243; eBiosciences, San Diego, CA, USA), anti-CD80 AF-647 (Clone MEM-233, AbD Serotec, Raleigh, NC, USA), anti-CD83 PE-Cy7 (Clone HB15e, eBiosciences), anti-CD86 AF-647 [Clone 2331 (FUN-1), BD; San Diego, CA, USA], anti CD40-APC-H7 (Clone 5C3, BD), anti-CD25 AF700 (Clone M-A251, BD), anti-CD69 PE-Cy7 (Clone FN50, Biolegend, San Diego, CA, USA), anti-CD134 PerCP-Cy5.5 (Clone ACT35, Biolegend), and Live/Dead ® Fixable Aqua Dead Cell Stain Kit (Invitrogen) for 30 min at room temperature in PBS containing 2% fetal calf-serum and 0.1% sodium azide.

Techniques: Activation Assay, Infection, Expressing