Journal: BioMed Research International

Article Title: Papain Ameliorates the MPAs Formation-Mediated Activation of Monocytes by Inhibiting Cox-2 Expression via Regulating the MAPKs and PI3K/Akt Signal Pathway

doi: 10.1155/2018/3632084

Figure Lengend Snippet: Effect of papain on the formation of monocyte-platelet aggregates and monocytes activation. (a) Gating strategy for the quantification of MPA subsets in the coculture system of THP-1 cells and activated platelets by flow cytometry (FCM). MPAs was evaluated by expression of CD14 and CD41. CD14 + cells suggested being monocytes, and activated platelet was identified by CD41 expression. (b) Surface marker of monocyte activation was measured by FCM detection. Activated monocytes were stained with CD11b. (c) Results of quantitative analysis of CD14 and CD41 as well as CD11b positive cells in different concentrations of papain groups. K-S test showed that data were normal distribution in all groups (P>0.05). ∗ indicates thrombin-treated group versus control group. # indicates papain and thrombin-treated group versus single thrombin-treated group. ∗∗ p<0.01, ## p<0.01.

Article Snippet: For the measurements of monocyte-platelet aggregates (MPAs), the mixture in each group was incubated with CD14-PE antibodies (rabbit anti-human monoclonal antibody, Bioss), CD41-APC antibodies (mouse anti-human monoclonal antibody, eBioscience), and isotype matched controls for 20 mins at room temperature.

Techniques: Activation Assay, Flow Cytometry, Expressing, Marker, Staining

Journal: BioMed Research International

Article Title: Papain Ameliorates the MPAs Formation-Mediated Activation of Monocytes by Inhibiting Cox-2 Expression via Regulating the MAPKs and PI3K/Akt Signal Pathway

doi: 10.1155/2018/3632084

Figure Lengend Snippet: Impact of TNF- α on the protective effect of papain on MPAs formation and monocytes activation. (a) Gating strategy for the quantification of MPA subsets in the coculture system of THP-1 cells and activated platelets by flow cytometry (FCM). MPAs were evaluated by expression of CD14 and CD41. CD14 + cells suggested being monocytes, and activated platelet was identified by CD41 expression. (b) Surface marker of monocyte activation was measured by FCM detection. Activated monocytes were stained with CD11b. (c) Results of quantitative analysis of CD14 and CD41 as well as CD11b positive cells in control and thrombin, papain, NS-398 and TNF- α -treated groups. K-S test showed that data were in normal distribution in all groups (P>0.05). ∗ indicates thrombin-treated group versus control group. # indicates papain and thrombin-treated or NS-398-treated groups versus single thrombin-only-treated group. ∧ indicates papain+TNF- α -treated group versus papain or NS-398-treated groups. ∗∗ p<0.01, ## p<0.01, ∧∧p<0.01.

Article Snippet: For the measurements of monocyte-platelet aggregates (MPAs), the mixture in each group was incubated with CD14-PE antibodies (rabbit anti-human monoclonal antibody, Bioss), CD41-APC antibodies (mouse anti-human monoclonal antibody, eBioscience), and isotype matched controls for 20 mins at room temperature.

Techniques: Activation Assay, Flow Cytometry, Expressing, Marker, Staining

Journal: Frontiers in immunology

Article Title: The significance of CD16+ monocytes in the occurrence and development of chronic thromboembolic pulmonary hypertension: insights from single-cell RNA sequencing.

doi: 10.3389/fimmu.2024.1446710

Figure Lengend Snippet: FIGURE 2 Changes in peripheral blood monocyte subsets in CTEPH patients. (A) UMAP plot of Mononuclear Phagocyte System (MPS) was divided into three cell types: monocytes, macrophages, and conventional dendritic cells (cDCs). Subsequently, monocytes were further divided into CD14+ monocytes and CD16+ monocytes. (B) A dot plot was used to annotate marker genes for CD14+ monocytes and CD16+ monocytes, along with a heatmap displaying the top 10 differentially expressed genes between the two cell types. (C) Ro/e (ratio of observed cell number to expected cell number) revealed the proportion of CD16+ monocytes in the CTEPH-N, CTEPH-I and HC groups. (D) The heat map showed the degree of correlation, and the numbers in the graph represented the correlation coefficient. (6MWD: 6-minute walk distance; PVR: pulmonary vascular resistance) (E) Schematic diagram of the gating strategy for distinguishing CD14+ monocytes from CD16+ monocytes by flow cytometry. (F) Differences in the proportion of CD16+ monocyte subsets in total monocytes between CTEPD patients (n = 15) and matched healthy controls (n = 15) were assessed by flow cytometry. All data were presented as means ± SEM, ***P < 0.001.

Article Snippet: Then the cells were resuspend in the flow cytometer wash buffer (2% FBS in PBS) and stained with the following antibodies according to the standard protocol: CD14-PE (Elabscience, E-ABF1209D) and CD16-FITC (Elabscience, E-AB-F1236C) for monocyte labeling.

Techniques: Marker, Cytometry

Journal: Frontiers in immunology

Article Title: The significance of CD16+ monocytes in the occurrence and development of chronic thromboembolic pulmonary hypertension: insights from single-cell RNA sequencing.

doi: 10.3389/fimmu.2024.1446710

Figure Lengend Snippet: FIGURE 3 The functional characteristics of peripheral blood CD16+ monocytes in CTEPH patients. (A) GO analysis (biological process) of upregulated genes in CD16+ monocytes compared with CD14+ monocytes in CTEPH patients. (B) KEGG analysis of upregulated genes in CD16+ monocytes compared with CD14+ monocytes in CTEPH patients. (C) GSEA bar plot of CD16+ monocytes versus CD14+ monocytes in patients with CTEPH. (D) GO analysis (biological process) of upregulated genes in CD16+ monocytes between the CTEPH and healthy control samples. (E) KEGG analysis of upregulated genes in CD16+ monocytes between CTEPH patients and healthy controls. (F) GSVA heatmap of CD16+ monocytes between CTEPH patients and healthy controls. (G) Heat map of transcription factors upregulated in CD16+ monocytes between CTEPH patients and healthy controls.

Article Snippet: Then the cells were resuspend in the flow cytometer wash buffer (2% FBS in PBS) and stained with the following antibodies according to the standard protocol: CD14-PE (Elabscience, E-ABF1209D) and CD16-FITC (Elabscience, E-AB-F1236C) for monocyte labeling.

Techniques: Functional Assay, Control

Journal: Atherosclerosis Plus

Article Title: Coronary atherosclerosis severity is closely associated with decreased GLP-1R positivity among CD16 + pro-inflammatory and patrolling monocyte subsets

doi: 10.1016/j.athplu.2021.10.001

Figure Lengend Snippet: Severe atherosclerosis is associated with increased proportions of intermediate (CD14+/CD16+ IMM) and non-classical (CD14-/CD16+ NC) monocytes and decreased frequency of GLP-1R positivity in these subsets. (A) Proportions of classical (CD14+/CD16+ CM), intermediate (CD14+/CD16+ IMM) and non-classical (CD14-/CD16 + NCM) subsets in total circulating monocyte amount of healthy controls (grey bars, n = 13) and patients with severe atherosclerosis (black bars, n = 10). ( B ) Frequency of monocytes expressing GLP-1R among different monocyte subsets in healthy controls (grey bars, n = 13) and patients with severe atherosclerosis (black bars, n = 10). Data are means ± SE.

Article Snippet: Briefly, blood was treated with human Fc-Receptor blocking solution (Biolegend, San Diego, CA), and subsequently stained with detection antibodies: anti human CD14 (Biogems, Westlake Village, CA); anti human CD16 (Biolegend, San Diego, CA); and either anti GLP-1R or mouse IgG2b isotype control (R&D Systems Minneapolis, MN).

Techniques: Expressing

Journal: Pediatric Rheumatology Online Journal

Article Title: Surface expression and genotypes of Toll- like receptors 2 and 4 in patients with juvenile idiopathic arthritis and systemic lupus erythematosus

doi: 10.1186/1546-0096-11-9

Figure Lengend Snippet: Representative flow cytometry histograms showing (a) TLR2 and (b) TLR4 expression on CD14 + monocytes. Blue filled histograms: isotype controls and green or red filled histogram TLR expression. Mean channel fluorescence intensity (MFI) derived from fluorescence histogram was used to study the level of cell surface TLR expression. Delta MFI (dMFI) was calculated as a subtraction and recorded as the MFI of the TLR2 or TLR4 antibody minus the MFI of the isotype-matched control antibody.

Article Snippet: For surface staining 100 μl of whole blood samples were incubated with the following anti-human primary antibodies for 15 minutes in the dark at 4°C: anti-CD14 monoclonal antibody (mAb; 10 μl per 100 μl blood, FITC or PE labelled, clone 8 G3; Diaclone, Besançon, France), anti-TLR2 mAb (10 μl, FITC labelled, clone HTA 125; eBioscience, San Diego, CA) or TLR4 mAb (10 μl, PE labelled, clone HTA 125; eBioscience, San Diego, CA).

Techniques: Flow Cytometry, Expressing, Fluorescence, Derivative Assay, Control

Journal: Pediatric Rheumatology Online Journal

Article Title: Surface expression and genotypes of Toll- like receptors 2 and 4 in patients with juvenile idiopathic arthritis and systemic lupus erythematosus

doi: 10.1186/1546-0096-11-9

Figure Lengend Snippet: (a) Mean fluorescence intensity (dMFI) of TLR2-expression on CD14 + monocytes of patients with JIA, SLE and healthy controls. (b) Mean fluorescence intensity (dMFI) of TLR4-expression on CD14 + monocytes of patients with JIA, SLE and healthy controls.

Article Snippet: For surface staining 100 μl of whole blood samples were incubated with the following anti-human primary antibodies for 15 minutes in the dark at 4°C: anti-CD14 monoclonal antibody (mAb; 10 μl per 100 μl blood, FITC or PE labelled, clone 8 G3; Diaclone, Besançon, France), anti-TLR2 mAb (10 μl, FITC labelled, clone HTA 125; eBioscience, San Diego, CA) or TLR4 mAb (10 μl, PE labelled, clone HTA 125; eBioscience, San Diego, CA).

Techniques: Fluorescence, Expressing

Journal: Pediatric Rheumatology Online Journal

Article Title: Surface expression and genotypes of Toll- like receptors 2 and 4 in patients with juvenile idiopathic arthritis and systemic lupus erythematosus

doi: 10.1186/1546-0096-11-9

Figure Lengend Snippet: (a) Mean fluorescence intensity (dMFI) of TLR2-expression on CD14 + monocytes of patients in various disease phases compared to TLR2-expression on monocytes of healthy controls. (b) Mean fluorescence intensity (dMFI) of TLR4-expression on CD14 + monocytes of patients in various disease phases compared to TLR4-expression on monocytes of healthy controls.

Article Snippet: For surface staining 100 μl of whole blood samples were incubated with the following anti-human primary antibodies for 15 minutes in the dark at 4°C: anti-CD14 monoclonal antibody (mAb; 10 μl per 100 μl blood, FITC or PE labelled, clone 8 G3; Diaclone, Besançon, France), anti-TLR2 mAb (10 μl, FITC labelled, clone HTA 125; eBioscience, San Diego, CA) or TLR4 mAb (10 μl, PE labelled, clone HTA 125; eBioscience, San Diego, CA).

Techniques: Fluorescence, Expressing