Journal: Journal of Immunotoxicology

Article Title: Is TGFβ as an anti-inflammatory cytokine required for differentiation of inflammatory TH17 cells?

doi: 10.1080/1547691x.2016.1193574

Figure Lengend Snippet: Figure 1. Colonies of proliferating CD4þ T-cells during in vitro human TH17 cell differentiation. For stimulation and expansion of the T-cells, polyclonal stimulators (includ- ing anti-CD3 and anti-CD28 antibodies) were used (see ‘‘Methods’’ section). Representative micro-images are shown with different magnifications.

Article Snippet: From each sample, CD4þ T-cells were isolated by magnetic activated cell sorting (MACS) using human CD4 MicroBeads (Miltenyi Biotec, London, UK), according to the manufacturer instructions.

Techniques: In Vitro, Cell Differentiation

Journal: Journal of Immunotoxicology

Article Title: Is TGFβ as an anti-inflammatory cytokine required for differentiation of inflammatory TH17 cells?

doi: 10.1080/1547691x.2016.1193574

Figure Lengend Snippet: Figure 2. IL-1b, IL-6, and IL-23 – but not TGFb – are required for optimal in vitro differentiation of human CD4þ TH17 cells. (A) Flow cytometric dot-plot analysis of intra- cellular IL-17 expression by CD4þ T-cells. Representative data of five independent experiments (i.e. PBMC from five different donors) are shown. (B) Columns represent mean percentage (±SEM) of T-cells expressing intracellular IL-17 in each culture condition. ns: Non-significant (*p < 0.05).

Article Snippet: From each sample, CD4þ T-cells were isolated by magnetic activated cell sorting (MACS) using human CD4 MicroBeads (Miltenyi Biotec, London, UK), according to the manufacturer instructions.

Techniques: In Vitro, Expressing

Journal: Journal of Immunotoxicology

Article Title: Is TGFβ as an anti-inflammatory cytokine required for differentiation of inflammatory TH17 cells?

doi: 10.1080/1547691x.2016.1193574

Figure Lengend Snippet: Figure 3. IFNc production by human T-cells differentiated in presence of IL-1b, IL-6, and IL-23 – with or without TGFb as well. (A) Flow cytometric analysis of intracellu- lar expression of IFNc by TH17 cells derived from CD4þ T-cells. (B) Mean expression levels (±SEM) from five independent experiments (i.e. PBMC from five different donors) using various culture conditions (i.e. different TGFb concentration and inhibitors of TGFb signaling pathways) are shown. ns: Non-significant (*p < 0.05).

Article Snippet: From each sample, CD4þ T-cells were isolated by magnetic activated cell sorting (MACS) using human CD4 MicroBeads (Miltenyi Biotec, London, UK), according to the manufacturer instructions.

Techniques: Expressing, Derivative Assay, Concentration Assay, Protein-Protein interactions

Journal: Journal of Immunotoxicology

Article Title: Is TGFβ as an anti-inflammatory cytokine required for differentiation of inflammatory TH17 cells?

doi: 10.1080/1547691x.2016.1193574

Figure Lengend Snippet: Figure 4. TGFb effects on Foxp3 expression by CD4þ T-cells differentiated under TH17 cell-promoting conditions. (A) Intracellular staining of Foxp3. (B) Columns are mean (±SEM) Foxp3 expression levels in five independent experiments (i.e. PBMC from five different donors) (*p < 0.05).

Article Snippet: From each sample, CD4þ T-cells were isolated by magnetic activated cell sorting (MACS) using human CD4 MicroBeads (Miltenyi Biotec, London, UK), according to the manufacturer instructions.

Techniques: Expressing, Staining

Journal: Journal of Immunotoxicology

Article Title: Is TGFβ as an anti-inflammatory cytokine required for differentiation of inflammatory TH17 cells?

doi: 10.1080/1547691x.2016.1193574

Figure Lengend Snippet: Figure 6. TH17 cell-derived cytokines; modulation by TGFb and blocking of TGFb signaling. Cytokine levels in supernatants of CD4þ T-cells differentiated toward TH17 states in the presence or absence of TGFb and/or inhibitors of signaling pathways. Data showed means± SEM from five independent experiments (i.e. PBMC from five different donors). ND: not detected (*p < 0.05).

Article Snippet: From each sample, CD4þ T-cells were isolated by magnetic activated cell sorting (MACS) using human CD4 MicroBeads (Miltenyi Biotec, London, UK), according to the manufacturer instructions.

Techniques: Derivative Assay, Blocking Assay, Protein-Protein interactions

Journal: Cell reports

Article Title: Bone Morphogenic Proteins Are Immunoregulatory Cytokines Controlling FOXP3 + T reg Cells

doi: 10.1016/j.celrep.2020.108219

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse CD4-cre , Taconic , Model#: 4196.

Techniques: Recombinant, Adjuvant, Magnetic Beads, Staining, Chromatin Immunoprecipitation, SYBR Green Assay, Knock-Out, Software

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Automated generation of gene-edited CAR T cells at clinical scale

doi: 10.1016/j.omtm.2020.12.008

Figure Lengend Snippet: Automated generation of TCRα/β-free CAR T cells (A) Schematic overview of the automated process on the CliniMACS Prodigy platform. Day 0, T cell selection and activation. Day 1, T cell transduction with CD19-CAR encoding lentiviral particles. Day 3, electroporation of transduced T cells with TRAC -targeting TALEN. Days 9–13, expansion of engineered T cells followed by TCRα/β depletion, harvesting, and cryopreservation. (B and C) Cell expansion and cell viability. Cell numbers (B) and cell viability (C) were determined using an automated cell counter (NucleoCounter). (D and E) Cellular composition. Representative samples were taken from the culture at indicated cell processing steps, and cellular composition was determined by flow cytometry (see ). Shown are the single data points and the averages of three runs. T, T cells; B, B cells; M, monocytes; G, granulocytes; NK, natural killer cells; NKT, natural killer T cells; Tn/Tscm, T cell naive or T stem cell memory; Tcm, T cell central memory; Tem, T cell effector memory; Teff, T cell effector; UT, untreated T cells. ∗, the depletion process started with 50% of T cells from the pre-depletion step.

Article Snippet: Then, buffy coats were connected to the tubing set, T cells labeled and magnetically isolated using CliniMACS CD4 reagent and CliniMACS CD8 reagent (Miltenyi Biotec).

Techniques: Selection, Activation Assay, Transduction, Electroporation, Flow Cytometry