Journal: Clinical and Experimental Immunology

Article Title: The influence of bone marrow-and synovium-derived mesenchymal stromal cells from osteoarthritis patients on regulatory T cells in co-culture

doi: 10.1111/cei.12122

Figure Lengend Snippet: (a) CD4+CD25+CD127− Treg percentages of total CD4+ T cells at d0. Magnetic separation of CD4+CD25+CD127− cells was performed with each circle representing a single culture. Percentages varied from less than 20% to over 70%. (b) Gating strategy for regulatory T-cells (CD127−/FoxP3+).

Article Snippet: PBMC were then separated into a mixture of CD4 + CD25 – and CD4 + CD25 + CD127 – cells using magnetic separation (CD4 + CD25 + CD127 dim/– regulatory T cell Isolation Kit II, LS and LD columns, MidiMACSTM separator, all from Miltenyi Biotec, Bergisch Gladbach, Germany).

Techniques:

Journal: BMC Bioinformatics

Article Title: The development of a comparison approach for Illumina bead chips unravels unexpected challenges applying newest generation microarrays

doi: 10.1186/1471-2105-10-186

Figure Lengend Snippet: Technical replication on subsequent array versions . (A) Experimental analysis for the T reg data set: FACS analysis and sorting windows of CD4 + CD127 low CD25 + T reg cells and CD4 + CD127 + CD25 - T conv cells ( left ). Expression of FOXP3 in the respective T cell subsets was assessed by flow cytometry ( middle ) and quantitative RT-PCR ( right ). Boxplots were used to compare the dynamic range of signal intensities on the arrays for (B) the T reg data set and (C) the whole blood data set. Only signals for the 8299 identical oligonucleotides were used. Technical replicates were checked both by principle component analysis based on the 100 most variable genes for (D) the T reg data set and (E) the whole blood data set as well as hierarchical cluster analysis (see Additional file ).

Article Snippet: CD4 + CD127 low CD25 + (T reg ) and CD4 + CD127 + CD25 - (T conv ) T cells were stained with CD4, CD25 and CD127 mAb (all from BD Pharmingen) and sorted on a FACSDiva cell sorter.

Techniques: Expressing, Flow Cytometry, Quantitative RT-PCR

Journal: Oncoimmunology

Article Title: Zoledronic acid inhibits NFAT and IL-2 signaling pathways in regulatory T cells and diminishes their suppressive function in patients with metastatic cancer

doi: 10.1080/2162402X.2017.1338238

Figure Lengend Snippet: PBMC from cancer patients (n = 10) were stained and analyzed by flow cytometry for; A) Treg (CD3+CD4+CD127−CD25+FOXP3+ of lymphocytes), B) NK cell (CD3−CD56+ of lymphocytes), γδ T cell (CD3+γδ+), and Conventional T cell (CD3+FOXP3−) frequencies before and after ZA treatment. Statistical analyses were performed on pooled data using Student's t test.

Article Snippet: Alternatively, Treg were sorted on a BD FACSJazz CD4 + CD127 − CD25 high (>95%) after CD4 enrichment (Miltenyi Biotech).

Techniques: Staining, Flow Cytometry

Journal: Oncoimmunology

Article Title: Zoledronic acid inhibits NFAT and IL-2 signaling pathways in regulatory T cells and diminishes their suppressive function in patients with metastatic cancer

doi: 10.1080/2162402X.2017.1338238

Figure Lengend Snippet: A) Enriched Treg from healthy blood donors (n = 6) were stimulated with IL-2 in the presence or absence of ZA for 3 days and analyzed for the expression of CD25 by flow cytometry. A representative histogram and accumulated data mean ± SEM are shown and statistical analysis was performed by Wilcoxon test. Each symbol represents a distinct healthy individual. B) Enriched Treg from healthy blood donors (n = 3) were stimulated with and without IL-2 in the presence or absence of ZA for 15, 30, and 90 minutes and analyzed for the expression of phosphorylated STAT5 by flow cytometry. Representative histograms and accumulated data mean ± SEM are shown and statistical analysis was performed by Two-way ANOVA. **p = 0.004, *p = 0.01. C) Enriched or sorted Treg (n = 6) were overnight stimulated with IL-2 and OKT3 in the presence or absence of ZA and stained for total TGFβ. A representative histogram and accumulated data are shown and statistical analyses were performed by Wilcoxon test.

Article Snippet: Alternatively, Treg were sorted on a BD FACSJazz CD4 + CD127 − CD25 high (>95%) after CD4 enrichment (Miltenyi Biotech).

Techniques: Expressing, Flow Cytometry, Staining

Journal: Oncoimmunology

Article Title: Zoledronic acid inhibits NFAT and IL-2 signaling pathways in regulatory T cells and diminishes their suppressive function in patients with metastatic cancer

doi: 10.1080/2162402X.2017.1338238

Figure Lengend Snippet: A) PBMC from cancer patients (n = 6) were stained and analyzed by flow cytometry for conventional (CD3+FOXP3−) T cell and NK cell proliferation (Ki67-positive cells) pre and one week post ZA administration. B) Correlation between percent circulating Treg and NK and T cell proliferation (Ki67). Statistical analyses were performed on pooled data using A) Wilcoxon test, and B) Pearson's correlation. PBMC from cancer patients (n = 10) were stained and analyzed by flow cytometry for the expression of C) FOXP3 and D) CD25. Representative histograms and pooled data are shown and Statistical analyses were performed using Student's t test.

Article Snippet: Alternatively, Treg were sorted on a BD FACSJazz CD4 + CD127 − CD25 high (>95%) after CD4 enrichment (Miltenyi Biotech).

Techniques: Staining, Flow Cytometry, Expressing