Journal: International Journal of Molecular Sciences

Article Title: Short-Chain Fatty Acids Augment Differentiation and Function of Human Induced Regulatory T Cells

doi: 10.3390/ijms23105740

Figure Lengend Snippet: Flow cytometric gating strategy for the phenotypic markers of human in vitro induced regulatory T cells (iTregs) generated from adult naïve CD4 + cells. ( A ) Flow cytometric gating strategy for lymphocytes, single cells, viable cells, CD4 + cells, and non-activated and activated iTregs generated from adult naïve CD4 + cells. ( B ) Representative dot plots showing the percentages of Foxp3 and CD39 gated on either non-activated (left panels) or activated (right panels) iTreg cells. ( C ) Representative dot plots showing the GITR, ICOS, PD-1, PD-L1, and CTLA-4 expression patterns on either non-activated or activated iTreg cells generated from adult naïve CD4 + cells with propionate. Data summarize thirteen independent experiments.

Article Snippet: The antibodies used included CD4 (PerCP-Vio700, 130-093-504, Miltenyi Biotec), CD39 (APC, 130-109-455, Miltenyi Biotec), Foxp3 (Alexa Fluor 488, 320212, BioLegend), GITR (PE, 130-092-895, Miltenyi Biotec), ICOS (VioBlue, 130-100-737, Miltenyi Biotec), PD-1 (APC/Cy7, 329922, BioLegend), PD-L1 (PE/Cy7, 329718, BioLegend), and CTLA-4 (APC, 349908, BioLegend).

Techniques: In Vitro, Generated, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Short-Chain Fatty Acids Augment Differentiation and Function of Human Induced Regulatory T Cells

doi: 10.3390/ijms23105740

Figure Lengend Snippet: Enhanced expression of phenotypic markers in activated cells generated from adult naïve CD4 + cells. Scattered dot plot with mean + SEM showing the percentage of Foxp3 + and CD39 + cells, and mean fluorescence intensity (MFI) of GITR, ICOS, PD-1, PD-L1, and CTLA-4 gated on non-activated (Non-A) and activated (A) cells generated from adult naïve CD4 + cells under various conditions. All cells were cultured in the presence of transforming growth factor β and interleukin-2, and additionally with control, acetate, butyrate, or propionate. Data summarize thirteen independent experiments. Wilcoxon matched-pairs rank test was used for statistical analysis. * p < 0.05, *** p < 0.001.

Article Snippet: The antibodies used included CD4 (PerCP-Vio700, 130-093-504, Miltenyi Biotec), CD39 (APC, 130-109-455, Miltenyi Biotec), Foxp3 (Alexa Fluor 488, 320212, BioLegend), GITR (PE, 130-092-895, Miltenyi Biotec), ICOS (VioBlue, 130-100-737, Miltenyi Biotec), PD-1 (APC/Cy7, 329922, BioLegend), PD-L1 (PE/Cy7, 329718, BioLegend), and CTLA-4 (APC, 349908, BioLegend).

Techniques: Expressing, Generated, Fluorescence, Cell Culture, Control

Journal: International Journal of Molecular Sciences

Article Title: Short-Chain Fatty Acids Augment Differentiation and Function of Human Induced Regulatory T Cells

doi: 10.3390/ijms23105740

Figure Lengend Snippet: Short-chain fatty acids (SCFAs) potentiate the generation of human transforming growth factor β (TGF-β)-induced Tregs in vitro. ( A ) Representative dot plots gated on live CD4 + CD3 + T cells showing the percentage of Foxp3 + (upper panel) and CD39 + (lower panel) cells induced after culturing under various conditions: control (interleukin-2 (IL-2) + TGF-β), acetate (IL-2 + TGF-β + acetate), butyrate (IL-2 + TGF-β + butyrate), and propionate (IL-2 + TGF-β + propionate). ( B ) GITR, ICOS, PD-1, PD-L1, and CTLA-4 expression of live CD4 + CD3 + T cells cultured under various conditions (see the legend to Panel A), as shown by representative histograms. ( C ) Scattered dot plots showing the mean and standard error of the mean (SEM) summarizing thirteen independent experiments for adults and twelve independent experiments for cord blood. Friedman test with Dunn’s multiple comparisons (post hoc) was used for statistical analysis. * p < 0.05, ** p < 0.01.

Article Snippet: The antibodies used included CD4 (PerCP-Vio700, 130-093-504, Miltenyi Biotec), CD39 (APC, 130-109-455, Miltenyi Biotec), Foxp3 (Alexa Fluor 488, 320212, BioLegend), GITR (PE, 130-092-895, Miltenyi Biotec), ICOS (VioBlue, 130-100-737, Miltenyi Biotec), PD-1 (APC/Cy7, 329922, BioLegend), PD-L1 (PE/Cy7, 329718, BioLegend), and CTLA-4 (APC, 349908, BioLegend).

Techniques: In Vitro, Control, Expressing, Cell Culture

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Successful ex vivo expansion of tumor infiltrating lymphocytes with systemic chemotherapy prior to surgical resection

doi: 10.1007/s00262-023-03500-9

Figure Lengend Snippet: Characteristics of TILs generated with or without chemotherapy. A Generated TILs were dominantly CD3+ T cells, and CD4+ T and CD8+ T cells ratio were shown (B and C, respectively), D double-positive (DP) and double-negative (DN) CD39/CD69, E expression of 4-1BB, F PD-1, and G TIGIT (n = 5: with chemotherapy, n = 15: without chemotherapy). Cells were stained with CD45RA and CCR7 to determine the percentage of memory phenotypes, H TEMRA (CD45RA+ CCR7−), I Naïve-like-phenotype T cells (CD45RA+, CCR7 +), J central memory (Tcm, CD45RA-, CCR7 +) and K effector memory (Tem, CD45RA-, CCR7-), in CD4 and CD8 populations (n = 5: with chemotherapy, n = 11: without chemotherapy). The Wilcoxon rank sum test was performed. n. s.: not significant

Article Snippet: The following Abs were used, anti-CD3-FITC (Biolegend, Cat# 300415), anti-CD4-PerCP (Biolegend, Cat# 344608), anti-CD8-V500 (BD Biosciences, Cat# 560774), anti-PD-1-BV421 (BD Biosciences, Cat#564323), anti-4-1BB-PE (Biolegend, Cat# 309804), anti-TIGIT-PE-Cy7 (Biolegend, Cat# 372714), anti-CD39-APC (BD Biosciences, Cat# 560239), anti-CD69-APC.Cy7 (BD Biosciences, Cat# 560737).

Techniques: Generated, Expressing, Staining

Journal: Stem Cells International

Article Title: Chondrogenic Potency Analyses of Donor-Matched Chondrocytes and Mesenchymal Stem Cells Derived from Bone Marrow, Infrapatellar Fat Pad, and Subcutaneous Fat

doi: 10.1155/2016/6969726

Figure Lengend Snippet: Immunoprofiles for MSC markers and putative chondrogenic potency markers of culture expanded cells prior to chondrogenesis. (a) ISCT MSC immunoprofiles. Immunoprofiles for the putative chondrogenic markers CD49c (b), CD166 (c), and CD39 (d). Flow cytometry was used to detect the percentage of positive cells for each marker on monolayer cell populations of chondrocytes (Ch), bone marrow MSC (BM), fat pad MSC (FP), and subcutaneous fat MSC (SCF) prior to chondrogenesis. Data shown are the means ± the standard deviation of 5 donors for each cell population. One-way ANOVA and post hoc Bonferroni tests were used to test for significant differences in the positivity of cell surface markers between cell types.

Article Snippet: CD markers which have been reported as putative chondrogenic potency markers [ – ] were also probed for; these included CD49c-PE (clone C3 II.1), CD166-BV421 (clone 3A6), CD39-APC (clone TU66), CD44-peridinin chlorophyll protein-cyanine 5.5 (PerCp-Cy5.5) (clone G44-26), and CD271-BV421 (clone C40-1457) (BD Biosciences).

Techniques: Flow Cytometry, Marker, Standard Deviation

Journal: Stem Cells International

Article Title: Chondrogenic Potency Analyses of Donor-Matched Chondrocytes and Mesenchymal Stem Cells Derived from Bone Marrow, Infrapatellar Fat Pad, and Subcutaneous Fat

doi: 10.1155/2016/6969726

Figure Lengend Snippet: Multilevel modelling.

Article Snippet: CD markers which have been reported as putative chondrogenic potency markers [ – ] were also probed for; these included CD49c-PE (clone C3 II.1), CD166-BV421 (clone 3A6), CD39-APC (clone TU66), CD44-peridinin chlorophyll protein-cyanine 5.5 (PerCp-Cy5.5) (clone G44-26), and CD271-BV421 (clone C40-1457) (BD Biosciences).

Techniques:

Journal: Cell Journal (Yakhteh)

Article Title: Helios, CD73 and CD39 Induction in Regulatory T Cells Exposed to Adipose Derived Mesenchymal Stem Cells

doi: 10.22074/cellj.2020.6313

Figure Lengend Snippet: Percentage of the CD39 + CD73 + in CD25 + FOXP3 + and CD25 - FOXP3 + T cells population subsequent to co-culturing with adipose-derived mesenchymal stem cells (ASCs). After five days culturing of naïve CD4 + T cells with ASCs, percentage of CD39 + CD73 + T cells in A. CD25 + FOXP3 + population and B. CD25 - FOXP3 + population was evaluated by flow cytometry method. The results illustrate mean ± SEM. **; P<0.01 compared to the control group.

Article Snippet: Flow cytometry analysis was performed with a FACSCalibur (BD Biosciences) using directly labeled monoclonal Abs (mAbs), PerCP mouse anti-human CD4, FITC mouse anti-human CD25, APC mouse anti-human CD45RA, PerCP-CYTM5.5 mouse anti-human CD73, APC mouse anti-human CD39, PE mouse anti-human FOXP3 (BD Biosciences), APC antimouse/ human Helios (Biolegend, Germany) and isotype control antibodies, all according to the manufacturers’ protocol.

Techniques: Derivative Assay, Flow Cytometry

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Naive CD8 + T Cells Expressing CD95 Increase Human Cardiovascular Disease Severity

doi: 10.1161/ATVBAHA.120.315106

Figure Lengend Snippet: CyTOF Antibody List.

Article Snippet: CD39 , A1 , 160Gd , Fluidigm , Fluidigm.

Techniques: Conjugation Assay