Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Fenofibrate Increases the Population of Non-Classical Monocytes in Asymptomatic Chagas Disease Patients and Modulates Inflammatory Cytokines in PBMC

doi: 10.3389/fcimb.2021.785166

Figure Lengend Snippet: HLA-DR expression in monocyte. The mean fluorescence intensity (MFI) of HLA-DR was determined in basal CD14pos cells after 20 h of T. cruzi lysate (Tc) stimulation or fenofibrate (Tc + Fen) treatment. Monocytes were selected based on FSC and SSC. After excluding doublets and debris, live cells were selected, monocytes were classified by CD14 positive staining. The mean fluorescence intensity (MFI) of HLA-DR was calculated both in total monocytes (A) . It shows the mean fluorescence intensity (MFI) of CD14pos/HLA-DRpos monocytes in healthy (HI) (B) , asymptomatic (Asy) (C) and chronic Chagas disease (CHD) patients (D) , where each patient is represented by a circle. The results are shown as the mean of the experiments ± SEM. These data were analyzed by fitting a mixed effect model with a Tukey post-hoc test.

Article Snippet: PBMC were stained with LIVE/DEADTM fixable dye (Invitrogen) at room temperature for 15 min and labeled with the following antibodies at 4°C for 30 min: CD14 (#E-AB-F1209C, Elabscience), CD16 (#E-AB-F1005M, Elabscience), HLA-DR (#E-AB-F1111H, Elabscience), and CCR2 (#357209, Elabscience).

Techniques: Expressing, Fluorescence, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Fenofibrate Increases the Population of Non-Classical Monocytes in Asymptomatic Chagas Disease Patients and Modulates Inflammatory Cytokines in PBMC

doi: 10.3389/fcimb.2021.785166

Figure Lengend Snippet: HLA-DR expression in T. cruzi stimulated and fenofibrate treated monocyte subpopulations. The mean fluorescence intensity percentage of HLA-DR + cells was determined in PBMC stimulated or not with T. cruzi lysate (Tc) and treated or not with fenofibrate (Tc + Fen) after 20 h, according to CD14 and CD16 expression. It shows the mean fluorescence intensity (MFI) of classical (CD14high/CD16neg) (A) , intermediate (CD14high/CD16pos) (B) and non-classical (CD14low/CD16pos) (C) monocytes with HLA-DR + expression. The results are shown as the mean of the experiments ± SEM. These data were analyzed by fitting a mixed effect model with a Tukey post-hoc test.

Article Snippet: PBMC were stained with LIVE/DEADTM fixable dye (Invitrogen) at room temperature for 15 min and labeled with the following antibodies at 4°C for 30 min: CD14 (#E-AB-F1209C, Elabscience), CD16 (#E-AB-F1005M, Elabscience), HLA-DR (#E-AB-F1111H, Elabscience), and CCR2 (#357209, Elabscience).

Techniques: Expressing, Fluorescence

Journal: Frontiers in Immunology

Article Title: Shuttling Tolerogenic Dendritic Cells across the Blood–Brain Barrier In Vitro via the Introduction of De Novo C–C Chemokine Receptor 5 Expression Using Messenger RNA Electroporation

doi: 10.3389/fimmu.2017.01964

Figure Lengend Snippet: The semi-mature phenotype and tolerogenic messenger RNA (mRNA) expression profile of tolerance-inducing dendritic cells (TolDCs) was not affected by mRNA electroporation.

Article Snippet: The phenotype of DCs was characterized using the following fluorochrome-labeled mouse antihuman monoclonal antibodies: anti-CD83-fluorescein isothiocyanate (Life Technologies), anti-CD80-phycoerythrin (BD Pharmingen), antihuman leukocyte antigen (HLA)-DR-peridinin chlorophyll (BD Biosciences), anti-CD86-fluorescein isothiocyanate (BD Pharmingen), and anti-CCR5-phycoerythrin-cyanin 7.

Techniques: Expressing, Electroporation

Journal: Critical Care

Article Title: TCR activation mimics CD127 low PD-1 high phenotype and functional alterations of T lymphocytes from septic shock patients

doi: 10.1186/s13054-018-2305-5

Figure Lengend Snippet: SPADE (Spanning-tree Progression Analysis of Density-normalized Events) analysis. T-cell phenotype was evaluated by using a SPADE algorithm based on the expression of different markers measured by flow cytometry on whole blood samples from patients with septic shock at day 3 after the onset of shock (D3, n = 17) and in healthy volunteers (HVs) (n = 14). CD4 + and CD8 + T cells from patients and donors were clustered in nodes on the basis of their similarities of expressions of CD127, CD38, HLA-DR, PD-1, FoxP3, and CD25. One tree containing four nodes was built for CD4 + T lymphocytes and one tree containing three nodes for CD8 + T cells. The medians of fluorescence intensity (MFIs) of the different markers for each node are represented on CD4 + and CD8 + T cells in patients and in HVs. Data are presented as Tukey boxplots. Node 4 of the CD4 + T cells, with a CD127 low CD25 high FoxP3 high phenotype, corresponds to regulatory T cells. Node 3 corresponding to CD127 low PD-1 high cells has been highlighted in both CD4 + and CD8 + T cells

Article Snippet: KRO anti-HLA-DR Ab was purchased from Becton Dickinson (San Jose, CA, USA) and FITC anti-CCR7 Ab from R&D Systems.

Techniques: Expressing, Flow Cytometry, Fluorescence

Journal: Critical Care

Article Title: TCR activation mimics CD127 low PD-1 high phenotype and functional alterations of T lymphocytes from septic shock patients

doi: 10.1186/s13054-018-2305-5

Figure Lengend Snippet: Occurrence of CD127 low PD-1 high T cells in an ex vivo model of T-cell receptor activation of purified T cells from healthy volunteers ( n = 9). Purified T cells were activated with anti-CD3/28 antibody-coated beads (αCD3/28, 1:1 bead-to-cell ratio) or anti-CD3 antibody-coated beads (αCD3, 1:1 bead-to-cell ratio) or not stimulated (NS) during 5 days. The percentage of CD127 low PD-1 high ( a ) as well as the percentage of HLA-DR ( b ) positive cells were measured by flow cytometry among CD4 + (left panel) and CD8 + (right panel) T cells. Data are presented as Tukey boxplots. Mann–Whitney paired tests were used to compare values between non-stimulated and activated conditions, * P <0.05, ** P <0.01, *** P <0.001

Article Snippet: KRO anti-HLA-DR Ab was purchased from Becton Dickinson (San Jose, CA, USA) and FITC anti-CCR7 Ab from R&D Systems.

Techniques: Ex Vivo, Activation Assay, Purification, Flow Cytometry, MANN-WHITNEY