Professor L.G.Rudenko

Belgrade, 26-27 March 2012

� WHO

� PATH (USA)

� CDC (USA)

� Serum institute of India (SII)

� BCHT (China)

� GPO (Thailand)

� BioDiem (Australia)

� Ongoing reconstruction of new facilities for Department of Virology.

� Agreement signed with NVI (Netherlands) for Proposed Preclinical study of H2N2 LAIV in ferrets

IMMUNOGENICITY OF H7N3 LAIV

IN FERRET MODEL

(HI data)

HI

tite

r

Weeks after vaccination

4 61

Weeks after vaccination

4 61

Placebo

H7N3 LAIV

Blood samples were collected from vaccinated animals on Day 0, 28 and 42 and

immunogenicity of the vaccine was evaluated in HI assay against homologues virus.

Vaccination – Day 0. Revaccination – day 28.

PROTECTIVE EFFICACY OF H7N3 LAIV

IN FERRET MODEL AFTER SUBSEQUENT

CHALLENGE WITH H7N3 WT VIRUS

Test articleFerret No

Virus titer, PFU/ml

Nasal washes Lungs

1 dpi 3 dpi 5 dpi7dpi

9dpi

3 dpi

H7N3 LAIV1–3 0 0 0

4–6 0 0 0 0 0

Placebo

7 8000 11000 0

8 3000 6000 75

9 30 110 0

10 0 1000 0 0 0

11 0 400 0 0 0

12 0 65 0 0 0

Animals were challenged on Day 42.

� Ongoing development agreements and brochure for initiation Phase I clinical trails A/17/turkey/Turkey/05/133 (H5N2) in Autumn 2012

� Preclinical studies of three candidates of LAIV H5 completed in IEM and University of Pittsburg. The best candidate -A/17/turkey/Turkey/05/133 (H5N2) selected for clinical trials

Test articleNo of

ferrets

Antigen H5N1 for HAI

Vietnam

(clade 1)

W/Swan

(clade 2.2)

Indonesia

(clade 2.1)

Anhui

(clade 2.1)

Bar–H Goose

(clade 2.2)

Wk 4 Wk 6 Wk 4 Wk 6 Wk 4 Wk 6 Wk 4 Wk 6 Wk 4 Wk 6

VN LAIV (clade 1) 12 44.9 359.2 11.9 33.7 11.9 13.3 10.0 17.8 10.0 20.0

Turkey LAIV (clade 2.2) 12 95.2 403.2 10.0 40.0 10.0 44.9 10.0 22.5 14.1 127.0

PBS 18 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0

IMMUNOGENICITY OF H5N2 LAIV

IN FERRET MODEL

(HI data)

PROTECTIVE EFFICACY OF H5N2 LAIV

IN FERRET MODEL AFTER SUBSEQUENT

CHALLENGE WITH A/TURKEY/TURKEY/1/2005

(H5N1) HPIV WT VIRUS (1)

Test articleFerret No

Virus titer, PFU/ml

Nasal washes Lungs

1 dpi 3 dpi 5 dpi7dpi

9dpi

3 dpi

H5N2 LAIV

809 0 1100 0

607 0 3100 0

609 0 900 0

604 0 2450 0 0 0

613 0 1900 0 0 0

614 0 950 0 0 0

Placebo

275 8000 534000 49500

277 3000 1250000 85500

270 30 825500 65000

453 1000 115000 32000 0 0

455 5500 325000 45000 0 0

456 15500 743000 24500 0 0Animals were challenged on Day 42.

Lesions score: none – 0; minimal – 1; slight – 2; moderate – 3; strong – 4; severe – 5.

Test articleMacroscopy lungs

lesions (score)*

Placebo 5, 5, 5, 5, 4, 4 (4.7)

NIBRG–23 (H5N1) PR8-based IIV against

A/turkey/Turkey/1/2005 (15 mkg HA in0.5 ml)2, 1, 1, 1, 1, 1 (1.2)

А/17/turkey/Turkey/05/133 (H5N2) LAIV (108 EID50 in 0.5 ml) 1, 0, 0, 0, 0, 0 (0.2)

A/17/duck/Potsdam/86/92 (H5N2) LAIV (108 EID50 in 0.5 ml) 3, 2, 1, 1, 0, 0 (1.2)

PROTECTIVE EFFICACY OF H5 VACCINES

IN FERRET MODEL AFTER SUBSEQUENT

CHALLENGE WITH A/TURKEY/TURKEY/1/2005

(H5N1) HPIV WT VIRUS (2)

H5N2 LAIVPLACEBO

PROTECTIVE EFFICACY OF H5N2 LAIV

IN FERRET MODEL AFTER SUBSEQUENT

CHALLENGE WITH H5N2 HPIV WT VIRUS (3)

Development and preclinical studies of new prepandemic vaccine candidate LAIV H2N2

• To generate H2N2 vaccine strains we chose one of the previously used

A(H1N1) 6:2 vaccine reassortants as a source of six internal genes of

A/Leningrad/134/17/57 (H2N2):

• A/17/New Caledonia/99/145 (H1N1).

• Full-genome sequencing of A/17/New Caledonia/99/145 (H1N1) strain

revealed two additional amino acid substitutions within its internal genes: in

PB1 gene (Met-317-Ile) and in NP gene (Leu-341-Ile). Both mutations are

attributable to more attenuated MDV A/Leningrad/134/47/57 (H2N2) and most

probably were transferred to the vaccine strain due to the heterogeneous

nature of uncloned A/Leningrad/134/17/57 virus. Both mutations could

potentially affect immunogenic properties of the vaccine strain.

• To avoid transfer of the additional mutations to H2N2 vaccine strains we

attempted to generate new A(H1N1) 6:2 reassortant using cloned master

donor virus A/Leningrad/134/17/K7/57 (H2N2) and A/New Caledonia/20/99

wild-type virus.

H2N2 LAIV Project

• Newly generated 6:2 reassortant virus A/K7/New Caledonia/99/513(H1N1) was also fully sequenced to confirm the absence of additionalunwanted mutations in its internal genes.

• Two sets of reassortment are ongoing:

• A/K7/New Caledonia/99/513 (H1N1) x A/California/1/66(H2N2)

• A/K7/New Caledonia/99/513 (H1N1) x A/Tokio/3/67(H2N2)

• A/New Caledonia/20/99-specific rat antiserum is used forselection of H2N2 antigens.

• All generated 6:2 reassortant clones will be fully sequencedto find the best vaccine candidate.

One scientist from Influenza Division of CDC take fellowship for reassortants development for back up laboratory

Lyophilized influenza master donor viruses A/Leningrad/134/17/57(K7) (H2N2) and B/USSR/60/69 were transfered from IEM to CDC

Master bank of cold adapted donor influenza A/Leningrad/134/17/57 (K7) was prepared and characterized in accordance with standard operating procedures specific for CDC.

Preparation of a new WHO recommended LAIV reassortant A/Victoria/361/2011 (H3N2) is in progress.

� Ongoing consultancy works for development LAIV in chicken eggs and possibility to use cell line for production

� One scientist visited SII for 2 weeks

� Due diligence dossier passed to BCHT for development and production of LAIV

� Three strains for seasonal LAIV and anti-serums transferred to BCHT

� One scientist worked at BCHT 5 weeks for teaching and preparation Master seed viruses

� Ongoing consultancy works for development of LAIV

� IEM confirmed the phenotypic and genotypic characteristics of GPO Fluvac H5