Fig. 8.

Oil-in-water adjuvants improve AdV5 uptake by ALDC, increase and prolong recombinant antigen expression and enhance antigen presentation. AF568-labelled AdV5 was mixed with PBS or an oil-in-water adjuvant, added to ALDC, cultured overnight and washed twice prior to analysis. (a) Confocal microscopy was used to count the number of intracellular virions per cell 4 h post-infection. (b and c) Three-dimensional reconstructions of representative samples of ALDC infected with AdV5-568 in PBS or adjuvant respectively 4 h post-infection. (d) GFP MFI of ALDC infected with AdV5-GFP in PBS or adjuvant 24 h post-infection. (e) Frequency of GFP-expressing ALDC over time following infection with AdV5-GFP in PBS or adjuvant. (f) Ag85A-specific IFN-γ ELISpot using AdV5-Ag85A or AdV5-GFP in PBS or adjuvant. ALDC were infected with AdV5 recombinants as described in Methods and cultured with MHC-matched CD4⁺ T cells from BCG-vaccinated animals. In all cases, the bars indicate means of 20 imaged cells from three different animals analysed in duplicate and error bars indicate standard error of the means.