Journal: Oncoimmunology

Article Title: Oncolytic measles virus induces tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity by human myeloid and plasmacytoid dendritic cells

doi: 10.1080/2162402X.2016.1261240

Figure Lengend Snippet: MV induces TRAIL expression and IFN-α secretion by pDCs and CD1c + DCs. pDCs were cultured with IL3, IL3+MV or the TLR7 agonist R837 (A). CD1c + DCs were cultured alone (−), with MV or R837 (B). The expression of surface markers by the indicated cells was determined by flow cytometry. IFN-α secretion was measured by ELISA. #, values are below the limit of detection of the kit (7 pg/mL). Results are expressed as the mean ± SEM of three independent experiments. * p < 0.05, one-way ANOVA (Kruskal–Wallis).

Article Snippet: DCs were also cultivated with the TLR7 agonist R837 (1 μg/mL, Invivogen), exogenous type I IFNs (rhIFN-α-2a and rhIFN-β-1a, 100 ng/mL, ImmunoTools) or the RIG-I agonist 5′-ppp-dsRNA LyoVec (10 μg/mL, Invivogen).

Techniques: Expressing, Cell Culture, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Oncoimmunology

Article Title: Oncolytic measles virus induces tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity by human myeloid and plasmacytoid dendritic cells

doi: 10.1080/2162402X.2016.1261240

Figure Lengend Snippet: TRAIL expression depends on IFN-α secreted by DCs exposed to MV. pDCs (A) and CD1c + DCs (B) were pretreated or not with Ruxolitinib (Rux) before exposure to IL3+MV or MV respectively, or before exposure to R837. The secretion of IFN-α was measured by ELISA. #, values are below the limit of detection of the kit (7 pg/mL). The expression of TRAIL by the indicated cells was determined by flow cytometry. (C) pDCs and CD1c + DCs were pretreated or not with Rux before exposure to type I IFNs (rhIFN-α-2a and rhIFN-β-1a). TRAIL positive cells were quantified by flow cytometry. Results are expressed as the mean ± SEM of three independent experiments. * p < 0.05, Mann–Whitney test.

Article Snippet: DCs were also cultivated with the TLR7 agonist R837 (1 μg/mL, Invivogen), exogenous type I IFNs (rhIFN-α-2a and rhIFN-β-1a, 100 ng/mL, ImmunoTools) or the RIG-I agonist 5′-ppp-dsRNA LyoVec (10 μg/mL, Invivogen).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, MANN-WHITNEY

Journal: Oncoimmunology

Article Title: Oncolytic measles virus induces tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity by human myeloid and plasmacytoid dendritic cells

doi: 10.1080/2162402X.2016.1261240

Figure Lengend Snippet: MV induces IFN-α and TRAIL expression following RLR activation in CD1c + DCs and RLR/TLR7 activation in pDCs. (A) pDCs were pretreated or not with IRS661 (TLR7 inhibitor) or with MRT67307 (TBK1 and IKK-ε inhibitor) and then cultured with IL3, IL3+MV or R837. (B) CD1c + DCs were pretreated or not with IRS661 or MRT67307 and then cultured alone (−) or with MV. (C) pDCs were cultured with IL3+MV or with IL3+UV-inactivated MV (MV UV*) and CD1c + DCs were cultured alone (−), with MV or MV UV*. (D) pDCs and CD1c + DCs were exposed to 5′-ppp-dsRNA LyoVec, a RIG-I agonist. (A–D) IFN-α secretion was measured by ELISA. #, values are below the limit of detection of the kit (7 pg/mL). The expression of TRAIL by the indicated cells was determined by flow cytometry. Results are expressed as the mean ± SEM of three independent experiments. (A, B) * p < 0.05, one-way ANOVA (Kruskal–Wallis). (C) * p < 0.05, Mann–Whitney test.

Article Snippet: DCs were also cultivated with the TLR7 agonist R837 (1 μg/mL, Invivogen), exogenous type I IFNs (rhIFN-α-2a and rhIFN-β-1a, 100 ng/mL, ImmunoTools) or the RIG-I agonist 5′-ppp-dsRNA LyoVec (10 μg/mL, Invivogen).

Techniques: Expressing, Activation Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, MANN-WHITNEY

Journal: Oncoimmunology

Article Title: Oncolytic measles virus induces tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity by human myeloid and plasmacytoid dendritic cells

doi: 10.1080/2162402X.2016.1261240

Figure Lengend Snippet: MV induces IFN-α and TRAIL expression after RIG-I activation in Gen2.2 cells. Gen2.2 cells established by transduction with lentiviruses expressing scrambled shRNA, shRNA against TLR7 (shTLR7) or RIG-I (shRIG-I) were cultured with IL3, IL3+MV or IL3+UV-inactivated MV (MV UV*). IFN-α secretion was measured by ELISA. #, values are below the limit of detection of the kit (7 pg/mL).The percentage of TRAIL positive Gen2.2 cells was determined by flow cytometry. Results are expressed as the mean ± SEM of three independent experiments. * p < 0.05, one-way ANOVA (Kruskal–Wallis).

Article Snippet: DCs were also cultivated with the TLR7 agonist R837 (1 μg/mL, Invivogen), exogenous type I IFNs (rhIFN-α-2a and rhIFN-β-1a, 100 ng/mL, ImmunoTools) or the RIG-I agonist 5′-ppp-dsRNA LyoVec (10 μg/mL, Invivogen).

Techniques: Expressing, Activation Assay, Transduction, shRNA, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Oncoimmunology

Article Title: Oncolytic measles virus induces tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cytotoxicity by human myeloid and plasmacytoid dendritic cells

doi: 10.1080/2162402X.2016.1261240

Figure Lengend Snippet: DCs activated with MV acquire a functional cytotoxic activity. (A) pDCs were pretreated or not with IRS661 or Rux and then cultured with IL3+MV, R837, type I IFNs (rhIFN-α-2a and rhIFN-β-1a) or 5′-ppp-dsRNA LyoVec. (B) CD1c + DCs were pretreated or not with IRS661 or Rux and then cultured with MV, UV-inactivated MV, R837, type I IFNs (rhIFN-α-2a and rhIFN-β-1a) or 5′-ppp-dsRNA LyoVec. (A, B) DCs were then added to the target Jurkat cells at an effector:target ratio of 20:1 and the percentage of specific lysis of Jurkat cells was determined by the measure of 51 Cr release in the supernatants. Results are expressed as mean ± SEM of at least three independent experiments and each condition was assessed in triplicates for each experiment. For the left panel: * p < 0.05, one-way ANOVA (Kruskal–Wallis); for the right panel: * p < 0.05, Mann–Whitney test.

Article Snippet: DCs were also cultivated with the TLR7 agonist R837 (1 μg/mL, Invivogen), exogenous type I IFNs (rhIFN-α-2a and rhIFN-β-1a, 100 ng/mL, ImmunoTools) or the RIG-I agonist 5′-ppp-dsRNA LyoVec (10 μg/mL, Invivogen).

Techniques: Functional Assay, Activity Assay, Cell Culture, Lysis, MANN-WHITNEY

Journal: The Journal of Experimental Medicine

Article Title: Myeloid heme oxygenase–1 regulates innate immunity and autoimmunity by modulating IFN-β production

doi: 10.1084/jem.20081582

Figure Lengend Snippet: Clinical and histological effects of polyI:C administration in HO-1 M-KO mice with EAE. (A) Control ( n = 9) and HO-1 M-KO ( n = 10) mice were treated with intraperitoneal injections of polyI:C or PBS on days 5 and 8 after infection. Data are representative of three independent experiments (means ± SEM of at least nine mice per group). (B) Spinal cord sections from immunized, polyI:C-treated HO-1 M-KO (a and c) and control (b and d) mice 28 d after infection were stained with H&E and LFB. Representative sections from two independent experiments with five mice per group are shown. Bars, 100 µM. (C) Immunized HO-1 M-KO mice were treated with intraperitoneal injections of polyI:C on days 5 and 8 after infection, and were either treated with PBS or with rhIFN-β starting from day 9 after infection and continuing over a period of 14 d. Data are representative of two independent experiments (means ± SEM of 10 mice per group).

Article Snippet: Induction of EAE was performed as previously described ( ) with 50 µg of rat MOG peptide in CFA containing a total of 1 mg H37Ra per mouse. rhIFN-β (Betaferon; Schering-Plough) was administered for 14 d using mini-osmotic pumps (model 2002; Alzet).

Techniques: Infection, Staining