Journal: Virology

Article Title: Induction of interferon-alpha and tumor necrosis factor-related apoptosis-inducing ligand in human blood mononuclear cells by hemagglutinin-neuraminidase but not F protein of Newcastle disease virus.

doi: 10.1006/viro.2002.1413

Figure Lengend Snippet: FIG. 2. Induction of TRAIL on human PBMC (A) and on gated CD14 monocytes (B, C) by NDV live or NDV-UV and effect of antiviral mAbs. (A) PBMCs were either untreated or treated with NDV or NDV-UV (20 HU/107 PMBC) in 2 ml culture medium for 24 h. UV treatment of the virus was for either 5 or 30 min. The cells were then stained with biotinylated anti-TRAIL polyclonal antibody followed by PE-labeled streptavidin. Bold lines represent staining with TRAIL antibody; dotted lines show background staining with naı¨ve control serum. (B) Histogram of the gated CD14 monocytes in PBMC. For this purpose, the cells were stained with FITC-labeled anti-CD14 mAb. Bold line represents staining with this mAb and the dotted line shows background staining with irrelevant isotype control FITC-labeled. (C) Effects of monoclonal antiviral antibodies on TRAIL expression by NDV-treated CD14 cells. Anti-HN (HN.B) and anti-F (Icii) mAb were incubated with NDV (20 HU) for 1 h at room temperature before the mixture was added to PBMCs. After 24 h incubation, similar stainings were performed as described for (A) and (B) and analyzed for gated monocytes. The percentage of TRAIL-positive cells in gate M1 is indicated.

Article Snippet: TRAIL molecules were detected by flow cytometry using biotinylated purified immunoglobulins from a polyclonal goat anti-human TRAIL serum (RD Systems, Wiesbaden, Germany) and streptavidin-PE (Pharmingen Becton Dickinson).

Techniques: Virus, Staining, Labeling, Control, Expressing, Incubation

Journal: Virology

Article Title: Induction of interferon-alpha and tumor necrosis factor-related apoptosis-inducing ligand in human blood mononuclear cells by hemagglutinin-neuraminidase but not F protein of Newcastle disease virus.

doi: 10.1006/viro.2002.1413

Figure Lengend Snippet: FIG. 3. IFN--inducing activity of fixed NDV-infected BHK cells in human PBMC. BHK cells were trypsinized and treated with NDV-Ulster (10 HU/106) for 1 h (for virus binding) or they were treated for 24 h at 37°C (for infection during 24 h). After incubation, the cells were washed twice in PBS and fixed on ice with 1% paraformaldehyde for 15 min and washed again with PBS to remove the paraformaldehyde. (A) The viral proteins expressed at the BHK cell surface were determined after virus binding (1 h) or after virus replication (24 h) by flow cytometry using mouse polyclonal anti-NDV serum or the mAbs anti-HN (HN.B) and anti-F (Icii) and the PE-conjugated anti-mouse IgG (see Materials and Methods). Dotted lines correspond to uninfected BHK cells treated in the same way. (B) Isolated PBMC (1 106) were incubated at 37°C for 1 or 24 h in 0.4 ml of RPMI–10% FCS with the NDV modified fixed BHK cells in increasing concentrations and the release of IFN- into the supernatant was determined by ELISA.

Article Snippet: TRAIL molecules were detected by flow cytometry using biotinylated purified immunoglobulins from a polyclonal goat anti-human TRAIL serum (RD Systems, Wiesbaden, Germany) and streptavidin-PE (Pharmingen Becton Dickinson).

Techniques: Activity Assay, Infection, Virus, Binding Assay, Incubation, Flow Cytometry, Isolation, Modification, Enzyme-linked Immunosorbent Assay