anti human cd16 pe cy7  (Biogems International)

Journal: The Journal of Headache and Pain

Article Title: Expression of miR-155 in monocytes of people with migraine: association with phenotype, disease severity and inflammatory profile

doi: 10.1186/s10194-024-01842-y

Figure Lengend Snippet: Percentage of events recorded for the different monocytic sub-populations among the three study groups. Legend: Panel A : percentage of M1 monocytes (CD80 +) sub-populations. Panel B : percentage of M2 monocytes (CD163 +) sub-populations. CM-MO: chronic migraine with medication overuse, EM: episodic migraine, HCs: healthy controls, classical: “classical” monocytes (CD14 + /CD16– expression), non-classical: “non classical-intermediate” monocytes (CD14 + /CD16 + expression), M1: pro-inflammatory “M1” monocytes (CD80 + expression), M2 : anti-inflammatory “M2” monocytes (CD163 + expression). Box-plot: the range between the upper and lower border of the box indicates the interquartile range (IQR), spanning from the 25 th to the 75 th percentile. Within the box, the line indicates the median, and the cross denotes the mean. The upper and lower whiskers extend to the maximum and minimum values, excluding outliers. Symbols positioned above the upper whisker represent outliers, defined statistically as values beyond the 75 th percentile plus 1.5 times the IQR. Kruskal–Wallis Test was used for intergroup comparisons

Article Snippet: Subsequently, they were incubated for 30 min at 4 °C in the dark with all of the following monoclonal antibodies: Peridinin Chlorophyll Protein Complex (PerCP)-conjugated anti-human CD14 (BD Biosciences, 20µL per 1 × 10 6 cells), R-phyco-erythrin-cyanine7 (PE-Cy7)-conjugated anti-human CD16 (BD Biosciences, 5µL per 1 × 106 cells), and R-phycoerythrin (PE)-conjugated CD163 (BD Biosciences, 5µL per 1 × 106 cells) or R-phycoerythrin (PE)-conjugated CD80 (BD Biosciences, 5µL per 1 × 106 cells).

Techniques: Expressing, Whisker Assay

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

Journal: Cell Reports Methods

Article Title: An autologous ex vivo model for exploring patient-specific responses to viro-immunotherapy in glioblastoma

doi: 10.1016/j.crmeth.2024.100716

Figure Lengend Snippet: Immunophenotyping and cytokine production capture OV-dose effects in the co-culture model Histograms depicting expression patterns for (A) CD69 and CD38 on CD8 + T cells in mock co-cultures and Delta24-RGD MOI 0.3- and MOI 30-infected co-cultures; (B) CD69 and CD38 on CD4 + T cells in mock co-cultures and Delta24-RGD MOI 0.3- and MOI 30-infected co-cultures; (C) CD69, CD38, HLA-DR, and CD16 on NK cells in mock co-cultures and Delta24-RGD MOI 0.3- and MOI 30-infected co-cultures; and (D) CD192, CD64, CD163, CD206, and Tim-3 on monocyte-derived macrophages in mock co-cultures and Delta24-RGD MOI 0.3- and MOI 30-infected co-cultures. (E) Bar graphs depicting mean production of IFNγ, TNF-α, CXCL9, CXCL10, and IL-6 in mock co-cultures and Delta24-RGD MOI 0.3- and MOI 30-infected co-cultures. Statistical significance between groups was evaluated by two-way ANOVA with correction for multiple comparisons using Sidak method (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.005, and ∗∗∗∗p < 0.001).

Article Snippet: PE-Cy7 conjugated mouse anti-human CD16 , BD Biosciences , Cat#560918; clone 3G8; RRID: AB_10563252.

Techniques: Co-Culture Assay, Expressing, Infection, Derivative Assay

Journal: Cell Reports Methods

Article Title: An autologous ex vivo model for exploring patient-specific responses to viro-immunotherapy in glioblastoma

doi: 10.1016/j.crmeth.2024.100716

Figure Lengend Snippet:

Article Snippet: PE-Cy7 conjugated mouse anti-human CD16 , BD Biosciences , Cat#560918; clone 3G8; RRID: AB_10563252.

Techniques: Staining, Virus, Recombinant, Expressing, Enzyme-linked Immunosorbent Assay, Cell Viability Assay, Software

Journal: EMBO Molecular Medicine

Article Title: A novel phosphocholine‐mimetic inhibits a pro‐inflammatory conformational change in C‐reactive protein

doi: 10.15252/emmm.202216236

Figure Lengend Snippet: A Confocal fluorescence microscopy of ADP‐activated platelets bound to HUVEC mono cell layers. pCRP‐Atto 594 (depicted in red) was added and incubated with and without C10M. Anti‐CD62P‐FITC antibody was used to detect the platelets (green). HUVEC nuclei were counterstained with DAPI (blue). pCRP colocalizes with platelets on the endothelial cells. C10M inhibits CRP binding to activated platelets. Scale bar 50 μm. B, C Quantification of ICAM‐1 (B) and VCAM‐1 (C) expression on pCRP*/mCRP‐activated HUVECs. ICAM‐1 and VCAM‐1 expressions were measured by flow cytometry. ADP‐stimulated platelets were added to each sample (except for “Control” and “pCRP”) and served as activated cell membranes for pCRP dissociation to pCRP*/mCRP. C10M inhibits the generation of pCRP*/mCRP, thereby reducing the expression of ICAM‐1 and VCAM‐1. Mean fluorescence intensity (MFI) results in flow cytometry are shown with results normalized to control, mean ± SEM. P values were calculated with ANOVA and Tukey's post‐hoc test. Biological replicates, n = 4. D, E Expression of integrin subunit αM (CD11b) in neutrophils (D) and CD14 + monocytes (E) was accessed by flow cytometry as described previously (Kiefer et al , ). Human whole blood was incubated with 25 μg/ml pCRP, 20 μM ADP, and C10M (molar ratio 1:100, pCRP:C10M), respectively. CD11b expression was analyzed by flow cytometry in neutrophils (CD16 + , SSC high) and monocytes (CD14 + , SSC low). Shown are scatter plots of MFI results in flow cytometry with results normalized to control, mean ± SEM. P values were calculated with ANOVA and Tukey's post‐hoc test. Biological replicates, n = 4. F, G ROS generation in whole blood detected in CD14 + monocytes (F) and neutrophils (G) by redox‐indicator dihydroethidium (DHE; 10 μg/ml). Blood samples incubated for 3 h at 37°C, 5% CO 2 with 50 μg/ml pCRP and mCRP, 20 μM ADP and C10M (molar ratio 1:100, pCRP:C10M), respectively. Control was left unstimulated and mCRP served as a positive control (Thiele et al , ). Cells were washed after red blood cell lysis and analyzed by flow cytometry. Shown are MFI results with results normalized to control, mean ± SEM. P values were calculated with ANOVA and Tukey's post‐hoc test. Biological replicates, n = 6. H pCRP*/mCRP dependent NETosis in isolated human neutrophils detected by confocal immunofluorescence microscopy. Isolated neutrophils incubated for 3 h at 37°C, 5% CO 2 with 100 μg/ml pCRP with and without PC:LPC liposomes (LP) and C10M (molar ratio 1:100, pCRP:C10M), respectively. Control was left unstimulated and 100 nM phorbol 12‐myristate 13‐acetate (PMA) served as a positive control. Cells were washed, fixed, and stained, and analyzed by confocal microscopy. Results are given as a ratio of NETing cells/all cells per ROI, with mean ± SEM. P values were calculated with ANOVA and Tukey's post‐hoc test. Biological replicates, n = 3. Source data are available online for this figure.

Article Snippet: Mouse anti‐human CD14 Pacific Blue™ (clone M5E2), anti‐human CD16 phycoerythrin‐cyanine 7 (PE‐Cy7, clone 3G8) and anti‐human HLA‐DR allophycocyanin (APC, clone TU36) antibodies were purchased from BD Pharmingen™, BD Biosciences.

Techniques: Fluorescence, Microscopy, Incubation, Binding Assay, Expressing, Flow Cytometry, Positive Control, Lysis, Isolation, Immunofluorescence, Staining, Confocal Microscopy

Journal: EMBO Molecular Medicine

Article Title: A novel phosphocholine‐mimetic inhibits a pro‐inflammatory conformational change in C‐reactive protein

doi: 10.15252/emmm.202216236

Figure Lengend Snippet:

Article Snippet: Mouse anti‐human CD14 Pacific Blue™ (clone M5E2), anti‐human CD16 phycoerythrin‐cyanine 7 (PE‐Cy7, clone 3G8) and anti‐human HLA‐DR allophycocyanin (APC, clone TU36) antibodies were purchased from BD Pharmingen™, BD Biosciences.

Techniques: Recombinant, Crystallization Assay, Staining, Software, Immunofluorescence, Western Blot