Journal: Gene therapy

Article Title: Measles virus selectively blind to signaling lymphocyte activation molecule as a novel oncolytic virus for breast cancer treatment.

doi: 10.1038/gt.2012.44

Figure Lengend Snippet: Figure 2. PVRL4 and not CD46 is involved in the infection of breast cancer cell lines with rMV-SLAMblind. (a) Surface expression of CD46 and PVRL4 in breast cancer cell lines analyzed by flow cytometry. (Left) Cells incubated with anti-CD46 mouse MAb (gray histogram) or isotype control (white histogram) followed by incubation with Alexa-488-conjugated goat anti-mouse antibody. (Right) Cells incubated with anti- PVRL4 goat polyclonal antibody (gray histogram) or isotype control (white histogram) followed by incubation with Alexa-488-conjugated rabbit anti-goat antibody. (b) Cells were pretreated with anti-CD46 or anti-PVRL4 antibody and infected with rMV-EGFP, rMV-EGFP-SLAMblind or rMV-Edmonston at an MOI of 0.1. Cells were incubated in medium with or without antibody and were photographed at 2 (Vero) or 3 d.p.i. (the others). Cells infected with rMV-Edmonston were immunostained using anti-N rabbit polyclonal antibody. Magnification: 100. (c) CHO- K1 cells were transfected with pCAG-hCD46 or the empty vector (pCAGGS). After 2 days, they were infected with rMV, rMV-SLAMblind or rMV- Edmonston at an MOI of 1 and incubated in the presence of fusion inhibitory peptide. At 2 d.p.i., cells were fixed and immunostained using anti-N MAb and the number of infected cells was counted. Infectivity in cells transfected with pCAGGS was set as 100%. Error bars indicate s.e. *Po0.001 versus pCAGGS. (d) BHK cells were transfected with pCAG-hPVRL4 or pCAGGS. After two days, they were infected with rMV, rMV- SLAMblind or rMV-Edmonston at an MOI of 0.1 and incubated in the presence of fusion inhibitory peptide. At 2 d.p.i., infectivity was determined as described in (c). *Po0.001 versus pCAGGS.

Article Snippet: Infection inhibition assay using antibodies Monolayers of MCF7, MDA-MB-453, SKBR3 and Vero cells in 96-well plates were pretreated for 1 h at 37 1C with medium containing 10mg ml 1 of anti-human CD46 MAb (clone M177; HyCult Biotechnology, Uden, The Netherlands)50 or anti-human PVRL4 goat polyclonal antibody (R&D Systems, Minneapolis, MN, USA).

Techniques: Infection, Expressing, Cytometry, Incubation, Control, Transfection, Plasmid Preparation

Journal: Gene therapy

Article Title: Measles virus selectively blind to signaling lymphocyte activation molecule as a novel oncolytic virus for breast cancer treatment.

doi: 10.1038/gt.2012.44

Figure Lengend Snippet: Figure 3. Infection of CD46-positive normal human cells. (a) Surface expression of CD46, PVRL4 and SLAM in NHDFs analyzed by flow cytometry as described in Figures 1c and 2a. (b, c) NHDFs were infected with rMV-SLAMblind or rMV-Edmonston at an MOI of 1. (b) Cells fixed and immunostained with anti-N MAb at 3 d.p.i. Magnification: 100. (c) Cell viability measured at each time point by WST-1 assay.

Article Snippet: Infection inhibition assay using antibodies Monolayers of MCF7, MDA-MB-453, SKBR3 and Vero cells in 96-well plates were pretreated for 1 h at 37 1C with medium containing 10mg ml 1 of anti-human CD46 MAb (clone M177; HyCult Biotechnology, Uden, The Netherlands)50 or anti-human PVRL4 goat polyclonal antibody (R&D Systems, Minneapolis, MN, USA).

Techniques: Infection, Expressing, Cytometry, WST-1 Assay