Immunogenic Profiling in Mice of a HIV/AIDS Vaccine Candidate (MVA-B) Expressing Four HIV-1 Antigens and Potentiation by Specific Gene Deletions
Figure 1
Characterization of MVA-B ΔA41L/ΔB16R recombinant virus.
(A) Scheme of MVA-B and MVA-B ΔA41L/ΔB16R genome maps, adapted from [13] and [57]. The different regions are indicated by capital letters. The right and left terminal regions are shown. Below each map, the deleted or fragmented genes are depicted as black boxes. In MVA-B ΔA41L/ΔB16R the deleted A41L and B16R genes are indicated. The HIV-1 Gag-Pol-Nef (from isolate IIIB) and gp120 (from isolate BX08) clade B sequences driven by the synthetic early/late (sE/L) virus promoter inserted within the TK viral locus (J2R) are indicated, adapted from [7]. (B) PCR analysis of A41L and B16R locus. 100ng of viral DNA extracted from DF-1 cells infected at 2 PFU/cell with MVA-WT, MVA-B or MVA-B ΔA41L/ΔB16R was used for PCR analysis. The DNA products corresponding to the parental virus or to the deletion are indicated by an arrow on the right, with the expected size in base pairs. Molecular size marker (1Kb ladder) with the corresponding sizes (base pairs) is indicated on the left. Lane Mock, cells not infected. (C) Expression of HIV-1 BX08gp120 and IIIBGPN proteins in DF-1 cells infected (2 PFU/cell) with MVA-B and MVA-B ΔA41L/ΔB16R, at 24h post-infection. Arrows on the right indicate the position of HIV-1 BX08gp120 and IIIBGPN proteins. (D) Virus growth of MVA-B and MVA-B ΔA41L/ΔB16R in infected (0.01 PFU/cell) DF-1 cells at different times and titrated by plaque immunostaining assay with anti-WR antibodies. The mean of three independent experiments are shown.