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 comparative neurology

Article Title: The distribution, number and certain neurochemical identities of infracortical white matter neurons in a lar gibbon ( Hylobates lar ) brain.

doi: 10.1002/cne.24545

Figure Lengend Snippet: Sources and dilution of antibodies used in the current study.

Article Snippet: nNOS , rabbit , Recombinant human neuronal nitric oxide synthase , Merck-Millipore , AB5380 , Russo et al., 2013 , 1:6000 , AB_91824.

Techniques: Recombinant

Journal: The Journal of comparative neurology

Article Title: The distribution, number and certain neurochemical identities of infracortical white matter neurons in a lar gibbon ( Hylobates lar ) brain.

doi: 10.1002/cne.24545

Figure Lengend Snippet: Stereological parameters used for estimating neuronal numbers in the white matter of the lar gibbon. NeuN – neuronal nuclear marker; nNos – neuronal nitric oxide synthase; CR – calretinin.

Article Snippet: nNOS , rabbit , Recombinant human neuronal nitric oxide synthase , Merck-Millipore , AB5380 , Russo et al., 2013 , 1:6000 , AB_91824.

Techniques: Marker, Sampling

Journal: The Journal of comparative neurology

Article Title: The distribution, number and certain neurochemical identities of infracortical white matter neurons in a lar gibbon ( Hylobates lar ) brain.

doi: 10.1002/cne.24545

Figure Lengend Snippet: Frequency distribution bar plots of somal volumes of WMICs in the brain of the lar gibbon. (a) Volumes of the soma of WMICs immunoreactive to neuronal nuclear marker (NeuN). Note the median volume is 615.9 μm3, with a range from 63.6 to 3 716.6 μm3. (b) Volumes of the soma of WMICs immunoreactive to neuronal nitric oxide synthase (nNOS). Note the median volume is 817.8 μm3, with a range from 110.6 to 2 899.8 μm3. (c) Volumes of the soma of WMICs immunoreactive to calretinin. Note the median volume is 619.4 μm3, with a range from 56.2 to 1 952.7 μm3. In all plots the bin width = 92.9 μm3.

Article Snippet: nNOS , rabbit , Recombinant human neuronal nitric oxide synthase , Merck-Millipore , AB5380 , Russo et al., 2013 , 1:6000 , AB_91824.

Techniques: Marker

Journal: The Journal of comparative neurology

Article Title: The distribution, number and certain neurochemical identities of infracortical white matter neurons in a lar gibbon ( Hylobates lar ) brain.

doi: 10.1002/cne.24545

Figure Lengend Snippet: Photomicrographs of neuronal nitric oxide synthase (nNOS) immunostaining in the rostral portion of the frontal lobe of the lar gibbon showing the distribution of WMICs immunoreactive to nNOS. (a) Moderately magnified image of the superior frontal gyrus (from the region indicated by the b in Fig. 1a), showing the numerous nNOS-immunoreactive WMICs deep to the cerebral cortex (grey matter, GM, white matter, WM). The approximate boundary of the deep border of cortical layer VI and the WM is marked by a dashed line. (b) High magnification image of the cortical/white matter boundary (marked by a dashed line) of the superior frontal gyrus, showing the nNOS-immunoreactive WMICs deep to the cerebral cortex. Arrows in a and b indicate the same neuron for orientation of image location. (c) Moderately magnified image of the fundus of the inferior frontal gyrus (from the region indicated by the d in Fig. 1a), showing the nNOS-immunoreactive WMICs deep to the cerebral cortex (GM) within the WM. The approximate boundary of the deep border of cortical layer VI and the WM is marked by a dashed line. (d) High magnification image of the cortical/white matter boundary (marked by a dashed line) of the fundus of the inferior frontal gyrus, showing the nNOS-immunoreactive WMICs deep to the cerebral cortex within the WM. Arrows in c and d indicate the same neuron for orientation of image location. Scale bar in c = 500 μm and apply to a and c. Scale bar in d = 250 μm and applies to b and d. In all images dorsal is to the top of the image and medial to the left.

Article Snippet: nNOS , rabbit , Recombinant human neuronal nitric oxide synthase , Merck-Millipore , AB5380 , Russo et al., 2013 , 1:6000 , AB_91824.

Techniques: Immunostaining