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. masque. Genetic differences between VZV wild-type and Oka strains Judith Breuer Professor of Virology St Barts and The London Hospital Medical School UK. 1. Rome, Italy - March 18, 2005. Oka vaccine Live attentuated virus. Bang BAN7. B. A. C. Bang +. J. 95. - PowerPoint PPT Presentation
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Genetic differences between VZV wild-
type and Oka strains
Judith BreuerProfessor of Virology
St Barts and The London Hospital Medical School
UK
1Rome, Italy - March 18, 2005
•masque
. . . .
Oka vaccine Live attentuated virus
A
B
CBang +
Bang BAN7
Reproduced from Barrett-Muir W, et al. J Virol 2002;76:1971-9with permission from the American Society for Microbiology
J95
Bang +
Bang BAN7
Loparev VN, et al. J Virol 2004;78: 8349-58.
E
M
J95
Bang +
Bang BAN7
Grose, J clin Virol 2005; in press
E
A
95
pOka vs Dumas
1/700
MSP vs Dumas
1/2000
BC vs Dumas
1/2000
BC vs
MSP
1/2700
Glycoproteins
5,14, 31, 37, 50, 60, 67 68
6 (8) 7 4 5(1)
transcriptional
4,10 21 61,62,63,64
6 0 1 1 (42)
replication
6, 8, 16, 18, 19, 28,29, 48,51,52,55,59
14 (18) 1 2 (2) 1
Kinases proteases
33,36, 47, 66
3 (2) 1 0 0
Tegument
11, 12,, 22, 54
9 (19) 3 4 4(1)
pOka vs Dumas
1/700
MSP vs Dumas
1/2000
BC vs Dumas
1/2000
BC vs
MSP
1/2700
pOka vs vOka-
1 in 2-3000
Glycoproteins
5,14, 31, 37, 50, 60, 67 68
6 (8) 9 6 5(1) 3(2)
transcriptional
4,10 21 61,62,63,64
6 0 1 1 (42) 10 (8)
replication
6, 8, 16, 18, 19, 28,29, 48,51,52,55,59
14 (18) 1 1 (2) 1 6 (1)
Kinases proteases
33,36, 47, 66
3 (2) 0 0 0 0 (1)
Tegument
11, 12,, 22, 54
9 (19) 3 4 4(1) 2 (1)
Which are the genetic correlates of vaccine attenuation?
Reduced ability of vOka to spread from T-cells to melanoma cell lines (Soong et al, 2000)
Similar to ORF 47 mutant
Evidence that ORF 47 p may be a virulence factor
Soong W, et al. J Virol. 2000;74:1864-70.
Vaccine preparations in comparison to pOka
6 9A 21 31 39 50 51 52 54 55 59 6 2
Nuc
leo
tide
po
siti
on
5745
1090
0
1277
9
3173
2
5859
5
7125
2
8730
6
8973
4
9053
5
9416
7
9774
8
1010
89
1053
10
1055
44
1057
05
1062
62
1067
10
1071
36
1072
52
1081
11
1088
38
O ka Parent A T C C A T T A A T G A A A T T A T T T A
O ka Biken G Y Y Y R Y Y G R C R R R G C C R C C C R
Nuc
leot
ide
posi
tion
5745
1090
0
1277
9
3173
2
5859
5
7125
2
8730
6
8973
4
9053
5
9416
7
9774
8
1010
89
1053
10
1055
44
1057
05
1062
62
1067
10
1071
36
1072
52
1081
11
1088
38
O ka Parent A T C C A T T A A T G A A A T T A T T T AO ka Biken G Y Y Y R Y Y G R C R R R G C C R C C C RO ka GSK MQO ka GSK MQO ka GSK AS
Vaccine preparations in comparison to pOka
Vaccine preparations in comparison to pOka
6 9A 21 31 39 50 51 52 54 55 59 6 2
Nuc
leot
ide
posi
tion
5745
1090
0
1277
9
3173
2
5859
5
7125
2
8730
6
8973
4
9053
5
9416
7
9774
8
1010
89
1053
10
1055
44
1057
05
1062
62
1067
10
1071
36
1072
52
1081
11
1088
38
O ka Parent A T C C A T T A A T G A A A T T A T T T AO ka Biken G Y Y Y R Y Y G R C R R R G C C R C C C RO ka GSK MQO ka GSK MQO ka GSK ASO ka GSK AS
Vaccine preparations in comparison to pOka
6 9A 21 31 39 50 51 52 54 55 59 6 2
Nuc
leot
ide
posi
tion
5745
1090
0
1277
9
3173
2
5859
5
7125
2
8730
6
8973
4
9053
5
9416
7
9774
8
1010
89
1053
10
1055
44
1057
05
1062
62
1067
10
1071
36
1072
52
1081
11
1088
38
O ka Parent A T C C A T T A A T G A A A T T A T T T AO ka Biken G Y Y Y R Y Y G R C R R R G C C R C C C RO ka GSK MQO ka GSK MQO ka GSK ASO ka GSK ASO ka Merk concensus
Fig. 1. Gel electrophoresis of polymerase chain reaction (PCR) products. A: Amplification across BssHII site in ORF 62 and restriction enzyme digest. Lane 1: Parental Oka virus; lane 2, UK wild type J virus; lane 3, Oka Merck vaccine; lanes 4–6, vaccine zoster viruses;lanes 7–18, vaccine rash viruses; lane 19, molecular weight marker (band sizes 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 kb).B: Amplification across Alu1 site in ORF 6 and restriction digest. Lane 1: 100 bp molecular weight marker; lane 2, parental Oka virus; lane 3, UK wild type J virus; lane 4, Oka Merck vaccine; lanes 5–7, vaccine zoster viruses; lanes 8–19, vaccine rash viruses; lane 20, 100 bp molecular weight marker.
Which mutations are important for attenuation?
Vaccine preparations in comparison to pOka
6 9A 21 31 39 50 51 52 54 55 59 6 2
Nuc
leot
ide
posi
tion
5745
1090
0
1277
9
3173
2
5859
5
7125
2
8730
6
8973
4
9053
5
9416
7
9774
8
1010
89
1053
10
1055
44
1057
05
1062
62
1067
10
1071
36
1072
52
1081
11
1088
38
O ka Parent A T C C A T T A A T G A A A T T A T T T AO ka Biken G Y Y Y R Y Y G R C R R R G C C R C C C RO ka GSK MQO ka GSK MQO ka GSK ASO ka GSK ASO ka Merk concensus
Is IE62 necessary for attenuation ?
Does the vaccine protect against wild type viruses?
IE 62 . Overlapping peptide pools
CD8 + cells assayed by Ifn staining
Peptide pool 5 recognised by A2 restricted PBLs
Conclusion
1. wtVZV varies geographically, and between related strains
2. vOka vaccine is attenuated biologically and clinically
3. The vOka vaccine is made up of a mixture of strains
4. Attenuation is likely to be multigenic and involve ORFs 30-54
5. vOka immune protection may be enhanced by the mixture of wt and vaccine mutations in the vaccine preparation.