Download 87. Induction of type-1 interferon by bovine plamacytoid dentritic cells (pDCs) in response to FMD virus

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 +
3.010 5
2.010 5
1.010 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