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Appendix 87 FMDV capsid: T independent and T dependent epitopes Antigen capture and presentation of FMDV by dendritic cells Antibody from immune cattle can form complexes with FMDV which allow the virus to enter, replicate in, and kill moDC 400 400 C ount C ount 2C2 500 300 4.88% 200 100 FMDV immune complexes significantly reduce T cell proliferative responses 500 400 300 4.18% 200 10 1 2 10 10 FL2-H 3 10 4 10 200 0 0 0 49.40% 300 100 100 0 10 O1K-Cad2 with immune serum (1/100) 2C2 O1K-Cad2 with nonimmune serum (1/100) 2C2 O1K-Cad2 alone 500 Count Depletion of CD4+ T cells during FMDV vaccination 0 10 1 2 10 10 FL2-H 3 10 4 10 0 10 1 2 10 10 FL2-H 3 10 4 Using IC to target inactivated FMDV to moDC results in significantly increased T cell stimulation “New tools and challenges for progressive control” Open Session of the EuFMD Research Group, Vienna (Austria) 29 September ‐ 1 October 2010 Appendix 21 Antigen targeting Viability of dendritic cells infected by recombinant viruses 4 3 10 10 10 10 4 10 3 2 1 10 10 10 0 0 256 512 10 768 1024 4 4 1834 3 1376 8.2% 2 459 0 0 0 256 Vector efficiency at delivering antigen to DCs in vitro? 0 102 F o rw a rd S c a tte r 10 10 10 10 3 10 2 10 1 10 0 10 0 256 512 768 1024 4 huAdV5 delivers antigen to more cells Adenovirus-infected cells express antigen more efficiently 10 10 10 3 2 1 10 10 10 0 0 256 512 10 768 1024 4 1319 2 1 256 512 10 10 10 3 2 1 0 256 512 0 10 768 1024 4 FSC GFP + 10 3 0 0 256 512 768 102424 0 10 10 1 2 10 3 10 3 38.9% 1846 2 1231 1 615 0 0 256 512 768 102424 FSC Viable cells 0 0 10 1 10 2 10 10 3 Propidium iodide Apoptotic cells rGated FSC CD205 T cell proliferation ALDCs + CD4 + 3.010 5 2.010 5 1.010 5 g -G F -F P M rF D V PV rF G PV FP -F M DV dv rA K rA dV 1u g 0 M 4 2461 high ia 10 Propidium iodide 4 MVA and Fowlpox kill dendritic cells ed 4 10 28.3% 367 0 10 2 10 734 1 10 1 10 Propidium iodide 1101 2 10 880 1468 A 3 10 0 4 10 4.2% Count 10 768 102424 4 10 3 10 0 0 FSC 4 2 10 440 0 FSC 10 1 10 Propidium iodide 3 Count 10 4 CD205 10 10 1759 FSC CD205 0.01 37.52 31.60 25.51 10 CD205 No Virus Adenovirus MVA Fowlpox Mean fluorescence (GFP) 109 570 172 257 rMVA-GFP % cells CD205 Recombina nt virus rFPV-GFP GFP+ 0 768 102424 4 Count 10 4 CD205 CD205 10 FSC 1u 768 BH 512 44 256 PW 0 rAdV-GFP 1 A- 10 M 10 2 512 FSC FSC 10 917 1 CPM FL4-CD205 10 No virus 10 CD205 10 GFP – marker of tropism Count (20 hours incubation) CD205 Infection of DCs with recombinant viruses in-vitro Antigen “New tools and challenges for progressive control” Open Session of the EuFMD Research Group, Vienna (Austria) 29 September ‐ 1 October 2010 4 10 Appendix 87 AdV5-GFP + Adjuvant – subcutaneously % of DCs GFP+ post inoculation Nick Juleff Bartek Bankowski Debi Gibson Julian Seago Lucy Robinson Kerry Mclaughlin Liz Reid Miriam Windsor Terry Jackson Paul Monaghan Pippa Hawes Ryan Waters Mark Woolhouse Ivan Morrison Adrian Hill Sarah Gilbert Andy Pollard Dave Stuart Liz Fry Claudine Porta “New tools and challenges for progressive control” Open Session of the EuFMD Research Group, Vienna (Austria) 29 September ‐ 1 October 2010