Journal: PLoS ONE

Article Title: Comparative Immunogenicity of HIV-1 gp160, gp140 and gp120 Expressed by Live Attenuated Newcastle Disease Virus Vector

doi: 10.1371/journal.pone.0078521

Figure Lengend Snippet: Twenty-four h post-infection, the infected cells were fixed with paraformadehyde and permeabilized with Triton X-100 for detection of total antigen inside the cell. The cells were probed with a pool of gp120-specific monoclonal antibodies followed by incubation with Alexa Fluor 488-conjugated goat anti-mouse IgG antibodies, and analyzed by immunofluorescence. The cells were visualized under Nikon Eclipse TE fluorescent microscope. Arrows indicate areas of positive immunofluorescence.

Article Snippet: The plates were washed three times with plate washing solution (Synbiotics Carporation) and incubated for 1 h with a 1:1,000 dilution of an isotype-specific secondary antibody; namely, horseradish peroxidase (HRP)-conjugated goat anti-guinea pig IgG (KPL, Gaithersburg, MD), goat anti-guinea pig IgG1, goat anti-guinea pig IgG2a (Novus Biologicals, Littleton, CO), or sheep anti-guinea pig IgA (Immunology Consultants Laboratory, Newberg, OR).

Techniques: Infection, Bioprocessing, Incubation, Immunofluorescence, Microscopy

Journal: PLoS ONE

Article Title: Comparative Immunogenicity of HIV-1 gp160, gp140 and gp120 Expressed by Live Attenuated Newcastle Disease Virus Vector

doi: 10.1371/journal.pone.0078521

Figure Lengend Snippet: The guinea pigs were immunized with the indicated rNDVs by the i.n. route. (A) The guinea pig sera were analyzed for NDV-specific antibodies by commercial NDV ELISA kits (Synbiotics Corporation). Mean ELISA end-point titers of NDV-specific serum antibodies on days 28 and 56 are shown. (B-D) The guine pig sera were analyzed by HIV-1 gp120 specific total IgG, IgG1 and IgG2 antibodies by isotype-specific ELISA with purified gp120. Mean ELISA end-point titers of gp120-binding serum antibodies of the indicated isotype on days 0, 7, 14, 21, 28, 42, 56, 70 and 90 are shown. Antibodies specific to gp120 were not detected in any animal on any day in the control rLaSota group. The graph shows the geometric mean value ± SEM for 3 animals in rLaSota, rLaSota/gp160 and rLaSota/gp140L groups, 6 animals in rLaSota/gp140S group and 5 animals in rLaSota/gp120 group. Arrows indicate time of rNDV immunizations on days 0 and 14. Statistical differences between the groups were calculated by unpaired t test (two-tailed). 1* indicates statistically significant differences ( P <0.05) of rLaSota/gp140S vs. rLaSota/gp160, rLaSota/gp140L and rLaSota/gp120 groups. 2* indicates statistically significant differences ( P <0.05) of rLaSota/gp140S vs. rLaSota/gp160 and rLaSota/gp140L groups.

Article Snippet: The plates were washed three times with plate washing solution (Synbiotics Carporation) and incubated for 1 h with a 1:1,000 dilution of an isotype-specific secondary antibody; namely, horseradish peroxidase (HRP)-conjugated goat anti-guinea pig IgG (KPL, Gaithersburg, MD), goat anti-guinea pig IgG1, goat anti-guinea pig IgG2a (Novus Biologicals, Littleton, CO), or sheep anti-guinea pig IgA (Immunology Consultants Laboratory, Newberg, OR).

Techniques: Enzyme-linked Immunosorbent Assay, Purification, Binding Assay, Control, Two Tailed Test