EFFICACY OF THE BACULOVIRUS BASED VACCINE FOR THE CONTROL OF THE H5N1 HPAI IN THE EGYPTIAN CONTEXT

Beato MS1, Bonfante F1, Buratin A1, Amarin N2 , Terregino C1, Gonzalez C2, Castellanos L2, Capua I1 , Realpe-Quintero Mauricio 2
1OIE/FAO and National Reference Laboratory for Avian Influenza and Newcastle disease OIE Collaborating Centre for Diseases at the Human-Animal Interface Istituto Zooprofilattico Sperimentale delle Venezie Padova - Italy 2 Research and Development Unit, Boehringer Ingelheim Vetmedica, Mxico

Novel vaccines
Only some epitopes of the AI virus is responsible for the neutralising antibody response
Induce protective immunity and reduce shedding levels

Detection of field exposure through antibodies to the NP, M, or N protein

Roles of Haemagglutinin (HA) protein

Characterisation of subtypes

Protection against infection

Pathogenicity

Subunit vaccines Baculovirus based

Powerful tool to apply the DIVA concept

Production in a timely and cost effective manner

No necessity to develop diagnostic compagnion tests (AGID, ELISA)

Subunit vaccines Baculovirus based

No licensed products available for poultry Regular update of the vaccine content Emergency vaccination

Vaccine strain selection for AI:criteria

Human field Veterinary field

Less harmonised
BUT
VACCINE BANK SUITABILITY OF BACULOVIRUS BASED VACCINE

Baculovirus based vaccines major challenges

Production of immunologically active proteins
Protein purification

Protein yield
Stability
Full length rHA generated from Human H1, H3 Avian H5, H7 (one study)

Hypothesis

Baculovirus based vaccine expressing the HA from clade 1 virus (A/duck/Chine/E319-2/03) Can induce protective immune response in SPF chickens against a highly virulent H5N1 HPAI strains

CIRCULATING IN EGYPT

Vaccines
Volvac BEST AI +ND KV

Vaccination scheme

10

31

Days of age
Bleeding
Volvac BEST AI +ND KV

Challenge study
Days of age
0 10 31

Days post infection

3 5 7 10 14

Immunisation

Tracheal and cloacal swab

Bleeding Infection

Bleeding

Bleeding: to assess post vaccination and post infection HI titres

Challenge viruses
A: rural poultry/human cases B: industrial poultry, more present in 2010 A/chicken/Egypt/1553-2/2010 subclade B A/chicken/Egypt/1709-6/2008 subclade B A/chicken/Egypt/3982-8/2010 subclade A

Challenge dose: 106 EID50/O.1 ml 6 experimental groups

Experimental set up
Volvac BEST AI +ND KV One dose

GROUP 1 A

GROUP 1 B

GROUP 1 C

A/CHICKEN/EGYPT/1553-2/2010

A/CHICKEN/EGYPT/1709-6/2008

A/CHICKEN/EGYPT/3982-8/2010

Survival rates
100

% OF HEALTHY ANIMALS

90 80 70 60 50 40 30 20 10 0 0 1 2 3 9 10 DAYS POST INFECTION 4 5 6 7 8 1A K 3982-8/10 K 1709-6/08 K 1553-2/10 1B 1C 1A

All birds of the control groups died within 5 days from infection (OIE requirements)

RESULTS- SEROLOGY
GROUP 1A
PRE CHALLENGE POST CHALLENGE

Log2 HI titres

10

Pre - H5BES Post - H5BES

Pre - NDV La Sota Post - NDV La Sota

Pre - A/chicken/Egypt/1553-2/10 Post - A/chicken/Egypt/1553-2/10

GROUP 1B
PRE CHALLENGE
12

POST CHALLENGE

Log2 HI titres

10

Pre - H5BES Post - H5BES

Pre - NDV La Sota Post - NDV La Sota

Pre - A/chicken/Egypt/3982-8/2010 Post - A/chicken/Egypt/3982-8/2010

GROUP 1C
PRE CHALLENGE POST CHALLENGE

Log2 HI titres

10

Pre - H5BES Pre - A/chicken/Egypt/1709-6/08 Post - NDV Lasota

Pre - NDV Lasota Post - H5BES Post - A/chicken/Egypt/1709-6/08

Viral shedding
Group 1 A: A/chicken/Egypt/1553-2/2010 SUBCLADE B

VIRUS DETECTION 3 day p.i. TS CS TS 5 day p.i. Virus CS isolation 7306/V10 6057/RS10 Not done Not done Not done Neg Not done Not done Not done Not done Not done Not done 0/10 Neg Neg Neg 26.07 Neg Neg Neg Neg Neg Neg 1/10 Virus isolation 7307/V10 Not done Not done Not done Pos Not done Not done Not done Not done Not done Not done 1/10 TS 6164/RS10 Neg Neg Neg 31.24 Neg Neg 23.75 27.22 Neg Neg 3/10 7 day p.i. Virus CS isolation 7309/V10 6165/RS10 Not done Not done Not done Neg Not done Not done Neg Neg Not done Not done 0/10 Neg Neg Neg 31.8 Neg Neg Neg Neg Neg Neg 1/10 10 day p.i. Virus isolation 7308/V10 Not done Not done Not done Pos Not done Not done Not done Not done Not done Not done 1/10 TS CS

Sample ID 6001/RS10 6002/RS10 6056/RS10 Animal ID 8 Neg Neg Neg 10 Neg Neg Neg 11 Neg Neg Neg 12 Neg Neg 35.34 13 Neg Neg Neg 14 Neg Neg Neg 15 Neg Neg Neg 16 Neg Neg Neg 17 Neg 37.2 Neg 19 Neg Neg Neg Total positive/Total 0/10 0/10 0/10 animals

6193/RS10 6194/RS10 Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/10 Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/10

Samples with a threshold cycle value (Ct) 35 were considered negative for influenza type A.

Viral shedding
Group 1B: A/chicken/Egypt/3982-8/2010 SUBCLADE A
VIRUS DETECTION 5 day p.i.

TS Sample ID 5741/RS10 Animal ID 1 32.49 3 Neg 4 32.29 5 Neg 6 Neg 7 Neg 8 33 9 33.85 10 33.95 12 34.9 Total positive/Total 6/10 animals

3 day p.i. Virus CS isolation 6974/V10 5742/RS10 Neg Not done Neg Not done Not done Not done Neg Neg Neg Neg 0/10 Neg 38.32 Neg Neg Neg Neg Neg Neg Neg Neg 0/10

Virus isolation 6975/V10 Not done Neg Not done Not done Not done Not done Not done Not done Not done Not done 0/10

10 day p.i. Virus TS CS TS CS TS CS isolation 5773/RS10 5774/RS10 5852/RS10 5853/RS10 5868/RS10 6976/V10 5869/RS10 Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/10 Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/10 Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/10 Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/10 Neg 35.6 Neg Neg Neg Neg Neg Neg Neg Neg 0/10 Not done Neg Not done Not done Not done Not done Not done Not done Not done Not done 0/10 Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/10

7 day p.i.

Samples with a threshold cycle value (Ct) 35 were considered negative for influenza type A.

Viral shedding
Group 1C: A/chicken/Egypt/1709-6/2008 subclade B

Sample ID Animal ID 14 15 16 17 23 24 26 28 29 30 Total positive/Tot al animals

VIRUS DETECTION 3 day p.i. 5 day p.i. 7 day p.i. 10 day p.i. TS CS TS CS TS CS TS CS 5489/RS1 5490/RS1 5534/RS1 5535/RS1 5600/RS1 5601/RS1 5624/RS1 5625/RS1 0 0 0 0 0 0 0 0 34 33.97 Neg Neg Neg Neg Neg Neg Neg Neg 2/10 Neg Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/10 D Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/9 D Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/9 D Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/9 D Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/9 D Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/9 D Neg Neg Neg Neg Neg Neg Neg Neg Neg 0/9

Samples with a threshold cycle value (Ct) 35 were considered negative for influenza type A.

CONCLUSIONS
All challenge viruses in nave birds caused death within 5 days and therefore the challenge requirements can be considered fulfilled as indicated by OIE Animals vaccinated with the Volvac BEST AI +ND KV exhibited high post vaccination HI titres

CONCLUSIONS Volvac BEST AI +ND KV

Confers protection against distinct viral variants Suitable for the Egyptian situation

Allows the application of the DIVA strategy

CONCLUSIONS

This is the first report available on the efficacy of a single-shot vaccination scheme using a whole baculovirus based vaccine expressing a haemagglutinin genetically distant from those of the challenge viruses

Control of AI
An universal solution for the prevention and control of AI does not exist

A combination of different strategies must be used on the basis of the characteristics of the poultry industry at risk
with goals that can be achieved within reason

Investigators
Study director: Dr. Ilaria Capua (IZSVe), Dr Carlos Gonzalez (BI)

Monitor: Dr. Calogero Terregino (IZSVe), Dr Mauricio Realpe (BI)

Study investigators: Maria Serena Beato; Francesco Bonfante, Cristian De Battisti Statisticians: Marzia Mancin, Marco Ruffa Laboratory technicians: Alessandra Buratin, Silvia Maniero, Silvia Ormelli, Angela Salomoni Animal handler: Serafino Pianta, Graziano Lando Quality assurance: Valeria Brasola