Journal: Neuroendocrinology

Article Title: Variation in the Dopamine-4-Receptor Gene in Patients with Type 1 Diabetes

doi: 10.1159/000530765

Figure Lengend Snippet: Frequency of CD14 ++ CD16 − and CD14 + CD16 + cells in patients with DM1 differing in the DRD4 −521C> T polymorphism

Article Snippet: 50 μL aliquots of fresh venous blood were stained with anti-CD14 (IgG2b mouse PerCP, clone MφP9; BD Biosciences, USA) and anti-CD16 (IgG2 mouse APC-Cy7, clone 3G8, BD Biosciences, USA) monoclonal antibodies.

Techniques:

Journal: PLoS ONE

Article Title: Differential Expression of CD163 on Monocyte Subsets in Healthy and HIV-1 Infected Individuals

doi: 10.1371/journal.pone.0019968

Figure Lengend Snippet: (A) Monocytes from whole blood were identified by forward and side scatter properties and expression of CD14 vs. CD16 with subsets defined as CD14++CD16− (P2, lower right), CD14++CD16+ (P3, upper right) and CD14+CD16++ (P4, upper left). (B) Expression of CD163 protein (dark grey) in each monocyte subset is shown compared to matched isotype control (light grey) in the histograms. Data illustrate results from a representative experiment. (C) CD163 mRNA was determined by Q-PCR in isolated monocyte subsets from 8 HIV-1 negative donors. Bars indicate median values. P values were calculated using the non-parametric, Mann-Whitney U test adjusted for multiple comparisons.

Article Snippet: An antibody cocktail composed of 8 µl anti-CD14 FITC or 4 µl anti-CD14 APC-Cy-7, 8 µl anti-CD16 PC5 and 8 µl CD163-PE (clone GHI/61; directed against domain 7 of CD163) or IgG-1 PE was placed in 5 ml polypropylene FACS tubes (BD Biosciences) and 80 µl of whole blood was added.

Techniques: Expressing, Isolation, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Differential Expression of CD163 on Monocyte Subsets in Healthy and HIV-1 Infected Individuals

doi: 10.1371/journal.pone.0019968

Figure Lengend Snippet: (A) Comparison of CD163 protein expression on the classical CD14++CD16− subset was carried out on 13 samples from HIV-1 negative donors and on all monocyte subsets from a further eight donors before and after Ficoll density gradient purification of PBMC from whole blood. Bars represent median values. Differences were analysed using the Wilcoxon matched pairs test. WB: whole blood, PBMC: peripheral blood mononuclear cells. (B) Whole blood from HIV-uninfected donors was incubated with 1 µg/ml LPS for 2 h and CD163 surface expression measured by flow cytometry. CD163 expression on each subset (mean fluorescence intensity) is plotted as the ratio of expression in treated compared to untreated monocytes. Data represent mean ± SEM for 4 independent donors. ∧ data from 1 donor was excluded from the CD14++CD16+ data set due to insufficient cell number.

Article Snippet: An antibody cocktail composed of 8 µl anti-CD14 FITC or 4 µl anti-CD14 APC-Cy-7, 8 µl anti-CD16 PC5 and 8 µl CD163-PE (clone GHI/61; directed against domain 7 of CD163) or IgG-1 PE was placed in 5 ml polypropylene FACS tubes (BD Biosciences) and 80 µl of whole blood was added.

Techniques: Expressing, Purification, Incubation, Flow Cytometry, Fluorescence

Journal: PLoS ONE

Article Title: Differential Expression of CD163 on Monocyte Subsets in Healthy and HIV-1 Infected Individuals

doi: 10.1371/journal.pone.0019968

Figure Lengend Snippet: Frequency of CD163 positive cells in CD14++CD16− monocytes (top row) and CD14++CD16+ monocytes (bottom row) was correlated with CD4 T cell count for donors below 500 cells/µl (left column) and above 500 cells/µl (right column). Donors not currently receiving antiretroviral therapy are represented by open circles and donors receiving protease inhibitors are represented by dotted squares. r s and P values after analysis excluding patients receiving PI are presented in parentheses after pooled data. r s value was determined by Spearman correlation coefficient.

Article Snippet: An antibody cocktail composed of 8 µl anti-CD14 FITC or 4 µl anti-CD14 APC-Cy-7, 8 µl anti-CD16 PC5 and 8 µl CD163-PE (clone GHI/61; directed against domain 7 of CD163) or IgG-1 PE was placed in 5 ml polypropylene FACS tubes (BD Biosciences) and 80 µl of whole blood was added.

Techniques: Cell Counting

Journal: Digestive Diseases and Sciences

Article Title: Regulation of the PD-1/PD-L1 Axis and NK Cell Dysfunction by Exosomal miR-552-5p in Gastric Cancer

doi: 10.1007/s10620-024-08536-0

Figure Lengend Snippet: Downregulation of peripheral blood NK cell-activating receptors NKG2D and NKp30 and cytokine factors IFN-γ and TNF-α in gastric cancer. A – C NK cell gating strategy. D and E Flow cytometry analysis comparing the expression of NK cell subsets CD3−/CD56+ and CD3−/CD16+ in GC and HD peripheral blood. F – H Flow cytometry analysis comparing the expression of NK cell receptors NKG2D, NKp30, and NKp46 in GC and HD peripheral blood. I and J ELISA detection of cytokine IFN-γ and TNF-α expression in GC (n = 16) and HD (n = 16) plasma. *P < 0.05; **P < 0.01; ***P < 0.001; ns not significant

Article Snippet: Single-cell suspensions were stained using the following antibodies: anti-human CD3 PE (BioGems, USA), anti-human CD16 PE-Cy7 (BioGems), anti-human CD56 APC (BioGems), anti-human NKG2D (CD314) PE (BioGems), anti-human NKp30(CD337) PE-Cy7 (BD, USA), anti-human NKp46(CD335) APC (BD), and PD-L1 PE (CST, USA).

Techniques: Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: Digestive Diseases and Sciences

Article Title: Regulation of the PD-1/PD-L1 Axis and NK Cell Dysfunction by Exosomal miR-552-5p in Gastric Cancer

doi: 10.1007/s10620-024-08536-0

Figure Lengend Snippet: Correlation of NK cell phenotype and activation receptor gene expression with prognosis in gastric cancer. Kaplan–Meier survival analysis revealed the following associations with poor prognosis in gastric cancer patients: low CD56 expression ( A ), low CD16 expression ( B ), low NKG2D expression ( C ), and low NKp46 expression ( E ). However, no correlation between NKp30 expression and prognosis was observed ( D )

Article Snippet: Single-cell suspensions were stained using the following antibodies: anti-human CD3 PE (BioGems, USA), anti-human CD16 PE-Cy7 (BioGems), anti-human CD56 APC (BioGems), anti-human NKG2D (CD314) PE (BioGems), anti-human NKp30(CD337) PE-Cy7 (BD, USA), anti-human NKp46(CD335) APC (BD), and PD-L1 PE (CST, USA).

Techniques: Activation Assay, Gene Expression, Expressing

Journal: Digestive Diseases and Sciences

Article Title: Regulation of the PD-1/PD-L1 Axis and NK Cell Dysfunction by Exosomal miR-552-5p in Gastric Cancer

doi: 10.1007/s10620-024-08536-0

Figure Lengend Snippet: Correlation of plasma exosomal miR-552-5p with NK cell phenotypes and activated receptors in gastric cancer. A and B Negative correlation between plasma exosomal miR-552-5p expression and the distribution of CD3−/CD56+ and CD3−/CD16+ subpopulations in peripheral blood NK cells of gastric cancer patients. C – E Negative correlation between plasma exosomal miR-552-5p expression and the expression of peripheral blood NK cell-activating receptors NKG2D, NKp30, and NKp46 in gastric cancer

Article Snippet: Single-cell suspensions were stained using the following antibodies: anti-human CD3 PE (BioGems, USA), anti-human CD16 PE-Cy7 (BioGems), anti-human CD56 APC (BioGems), anti-human NKG2D (CD314) PE (BioGems), anti-human NKp30(CD337) PE-Cy7 (BD, USA), anti-human NKp46(CD335) APC (BD), and PD-L1 PE (CST, USA).

Techniques: Clinical Proteomics, Expressing