T and B cell responses following immunization.

<p>B10.BR mice were vaccinated I.M. with Ad-CMVZGP (1×10⁵, 1×10⁶ and 1×10⁷ IFU/mouse) or Ad-CAGoptZGP (1×10⁴, 1×10⁵ and 1×10⁶ IFU/mouse) and splenocytes were harvested 8 days later for A. IFN-γ CD8+ T cells frequency analysis or B. Neutralizing antibody (NAB) titers. Four to five mice were analyzed per group and the experiment was repeated twice. Levels of NAB to ZEBOV-EGFP were evaluated 25 days post-vaccination. Error bars represent the standard deviation of the data. n.d. refers to assays that were not done for Ad-CAGoptZGP at that IFU/mouse. * p = 0.0454; ** p = 0.1.</p

T cell frequency analysis at day 6 post-immunization.

<p>Groups of 4 B10.BR mice were vaccinated I.M. with 1×10⁸ IFU/mouse of Ad-CMVZGP or Ad-CAGoptZGP and splenocytes were harvested 6 days later, re-stimulated with the peptide TELRTFSI, and production of IFN-γ, TNF-α, and Il-2 from CD8+ T cells was monitored by FACS. Error bars represent the standard deviation of the data. The experiment was repeated once and showed similar results. * p<0.001; ** p<0.001; *** p<0.01; **** p<0.01.</p

Survival and weight loss of mice challenged with ZEBOV 28 days post-vaccination.

<p>Eight to nine B10.BR mice were used in each group.</p>1<p>not applicable.</p

Western blot expression analysis of Ad-CMVZGP or Ad-CAGoptZGP.

<p>Proteins isolated from infected HEK 293 cells were separated by a 10% SDS PAGE then transferred to PVDF membrane. A mouse monoclonal anti-ZGP was used as the primary antibody and a goat anti-mouse horseradish peroxidase (HRP) conjugated antibody as the secondary antibody. 24, 48 and 72 hours indicate the time of total protein harvest post-infection. Band density corresponding to each lane is shown as determined by densitometry of the bands. The control represents untreated HEK 293 cells. An M.O.I. of 5 was used to infect the cells with either Ad-CMVZGP or Ad-CAGoptZGP for each time point. The preparation of Ad-CAGoptZGP or Ad-CMVZGP used had a non-infectious to infectious ratio of 73∶1or 19∶1 respectively.</p