Journal: Mediators of Inflammation

Article Title: Cytomegalovirus Infection Impairs Immunosuppressive and Antimicrobial Effector Functions of Human Multipotent Mesenchymal Stromal Cells

doi: 10.1155/2014/898630

Figure Lengend Snippet: CMV blocks IDO activity and IDO-mediated antimicrobial effects in human MSC. (a) PBL, stimulated with OKT3, were cocultured with MSC in the presence or absence of CMV. As controls UV-inactivated CMV (uvCMV), the IDO-specific inhibitor 1-L-methyl-tryptophan (1-MT; 1.5 mM), or a neutralising anti-IFN- γ antibody ( α IFN- γ ; 10 ng/mL) was used. After three days IDO activity was determined and is presented as mean kynurenine content ± SEM of three independent experiments, each done in triplicate. (b) PBL (1 × 10 5 /well), stimulated with CD3-directed mAB OKT3, were cocultured with MSC (3 × 10 4 /well) in the absence or presence of CMV (MOI 5). After three days cultures were infected with S. aureus (10–100 cfu/well) and bacterial growth was determined photometrically. As a control, cultures were supplemented with L-tryptophan (Trp; 0.6 mM) at the time point of bacterial infection. Data are given as mean OD (620 nm) ± SEM of three experiments, each done in triplicate. Significant differences ( P < 0.05) as compared to the positive control are marked by asterisks.

Article Snippet: Recombinant human IFN- γ was purchased from R&D Systems (Wiesbaden, Germany).

Techniques: Activity Assay, Infection, Control, Positive Control

Journal: Mediators of Inflammation

Article Title: Cytomegalovirus Infection Impairs Immunosuppressive and Antimicrobial Effector Functions of Human Multipotent Mesenchymal Stromal Cells

doi: 10.1155/2014/898630

Figure Lengend Snippet: CMV inhibits IDO induction by recombinant IFN- γ . (a) MSC (2 × 10 4 /well) which were infected with various amounts of CMV (MOI 0.1–10) were stimulated with IFN- γ (300 U/mL). After three days IDO activity was determined and is presented as mean kynurenine content ± SEM of three independent experiments, each done in triplicate. (b) MSC (1.5 × 10 6 /flask) were stimulated with IFN- γ (600 U/mL) in the absence or presence of CMV (MOI 5). Cells were harvested after 24 h and IDO protein was detected in Western blot analysis. β -Actin was utilized as a protein loading control, while the viral pp72 protein served as an infection control.

Article Snippet: Recombinant human IFN- γ was purchased from R&D Systems (Wiesbaden, Germany).

Techniques: Recombinant, Infection, Activity Assay, Western Blot, Control

Journal: Stem Cells Translational Medicine

Article Title: Safety Profile of Good Manufacturing Practice Manufactured Interferon γ‐Primed Mesenchymal Stem/Stromal Cells for Clinical Trials

doi: 10.1002/sctm.16-0485

Figure Lengend Snippet: Residual interferon γ in cell washings and Infusion medium a

Article Snippet: Two days before the expansion was complete, cells were fed with fresh D4 culture media containing 500 U/ml GMP‐grade recombinant human IFNγ (R&D Systems, Minneapolis, MN, http://www.rndsystems.com ).

Techniques: Control

Journal: Stem Cells Translational Medicine

Article Title: Safety Profile of Good Manufacturing Practice Manufactured Interferon γ‐Primed Mesenchymal Stem/Stromal Cells for Clinical Trials

doi: 10.1002/sctm.16-0485

Figure Lengend Snippet: Clustering of global transcriptional responses of MSCs following priming with IFNγ. (A): Principal Component Analysis (PCA) of differential gene expression of MSCs (open shapes) and γMSC (filled shapes) from each donor (same color). (B): Euclidean distance between samples to assess overall similarity among the samples. The darker blue indicates a shorter distance, more similar; lighter blue indicates greater distance, less similar. (C): Heat map analysis of RNA‐Seq gene expression data from MSCs and γMSC. The heatmap was generated using the normalized expression values for the 20 most upregulated and downregulated genes following interferon γ priming. Color code indicates relative expression; red highest, blue lowest. Abbreviation: MSCs, mesenchymal stem/stromal cells.

Article Snippet: Two days before the expansion was complete, cells were fed with fresh D4 culture media containing 500 U/ml GMP‐grade recombinant human IFNγ (R&D Systems, Minneapolis, MN, http://www.rndsystems.com ).

Techniques: Gene Expression, RNA Sequencing, Generated, Expressing

Journal: Stem Cells Translational Medicine

Article Title: Safety Profile of Good Manufacturing Practice Manufactured Interferon γ‐Primed Mesenchymal Stem/Stromal Cells for Clinical Trials

doi: 10.1002/sctm.16-0485

Figure Lengend Snippet: Soft agar colony forming assay to detect malignant transformation. Representative photographs of triplicate agar plates for each condition are shown. The positive control was a human ES‐2 ovarian clear cell carcinoma cell line. The negative control was saline. Original magnification was ×46. Abbreviations: IFNγ, interferon γ; MSCs, mesenchymal stem/stromal cells.

Article Snippet: Two days before the expansion was complete, cells were fed with fresh D4 culture media containing 500 U/ml GMP‐grade recombinant human IFNγ (R&D Systems, Minneapolis, MN, http://www.rndsystems.com ).

Techniques: Transformation Assay, Positive Control, Negative Control, Saline

Journal: Frontiers in Pharmacology

Article Title: Biotechnologically Produced Lavandula angustifolia Mill. Extract Rich in Rosmarinic Acid Resolves Psoriasis-Related Inflammation Through Janus Kinase/Signal Transducer and Activator of Transcription Signaling

doi: 10.3389/fphar.2021.680168

Figure Lengend Snippet: Schematic representation of the proposed mechanisms of action of Lavandula angustifolia extract (LV) and rosmarinic acid (RA) in psoriasis-like inflammation model in human keratinocytes. HaCaT cells exposed to IFN-γ/IL-17A/IL-22 respond with activation of JAK2/STAT1 and PI3K/AKT signaling pathways. Activation of JAK2 upon cytokine stimulation leads to STAT1 activation and its subsequent phosphorylation. Following nuclear translocation the phosphorylated STAT1 induces transcriptional activation of psoriasis-related inflammatory genes ( e.g., IL6 , CCL20 , CCL2 ). Activated PI3K/AKT axis in psoriatic keratinocytes correlates with induction of hyperproliferation and aggravation of the inflammatory milieu. In the present investigation, both RA and LV inhibited JAK2 and diminished STAT1 phosphorylation, hence, preventing inflammatory activation. Additionally, the LV extract disrupted PI3K/AKT signaling which could contribute to decrease in proliferation rate in activated keratinocytes. Taken together, the obtained data suggest that LV and RA possess inhibitory effect on psoriasis-related inflammation in human keratinocytes.

Article Snippet: The recombinant human IL-17A (#ENZ-PRT188) and IL-22 (#ENZ-PRT250) were purchased from Enzo Life Sciences AG (Lausen, Switzerland) and IFN-γ (#10067-IF) was obtained from R&D Systems (Minneapolis, MN, United States).

Techniques: Activation Assay, Translocation Assay