Workpackage 4

 

Work package number

4

Start date or starting event:

M19

Work package title

Immunogenicity studies in ferrets

Activity Type[1]

RTD

Participant number

1

4

5

 

 

 

 

Participant short name

iQur

UCL

CRP-Santé

 

 

 

 

Person-months per participant:

2.0

3.0

1.1

 

 

 

 

                   

 

Objectives

Short-listed vaccine candidates will be further tested in the ferret IAV model.

 

Description of work (possibly broken down into tasks), and role of participants

 

The ferret is generally considered as the best animal model for investigating human influenza infections, due to the similarity in the clinical course and pathological lesions of the infection in both species. Work will be coordinated by iQur and subcontracted on a best-buy for money basis to a GLP accredited contract research organisation in order to ensure compliance with clinical trial legislation. In principle, despite the differences between influenza subtypes these experiments will be limited to the most distant IAV subtypes successfully tested in mice.

 

Task 4.1: Expression of lead candidate (UCL, IQUR, M19-M20)

The optimal VLP construct will be chosen on the basis of the immunogenicity and efficacy studies in mice (WP 3). The VLP will be produced by UCL at small (5 litre) scale (approximately 2g). Purification will be carried out using the methodology developed in WP 1.5. The purified candidate VLP will be characterized (i) by ELISA (antigenicity, conformation) (ii) Western blot (purity) (iii) for stability (see WP2) and (iv) to confirm its immunogenicity in mice. 

 

Task 4.2: Protection from lethal challenge (IQUR (s/c), M19-M21)

Ferrets will be immunised sub-cutaneously using the optimal dosing strategy developed in WP 3.1. Serum will be taken from immunised ferrets to confirm seroconversion.  At this point, the animals will be challenged with a lethal (5xLD50) dose of a homologous seasonal IAV. Virus replication (nasal swabs), and survival will be monitored. At the end of the experiment, surviving animals will be sacrificed to test for immunohistochemistry and virus titres in the lung. Protection against lethal IAV infection should be reproducibly 100% after vaccination with the optimal VLP. If protection is incomplete another short-listed candidate will be tested or another round of optimization will be initiated.

 

Task 4.3. Efficacy testing against heterologous subtypes (IQUR (s/c), M22-M24)

If efficacy of the candidate vaccine against the homologous challenge can be confirmed, animals will be challenged with heterologous seasonal subtypes including pandemic H1N1 (2009).  If animals are protected against sequential heterologous challenges, a group of animals will be vaccinated and challenged with the most “distant” heterologous subtype.

 

 

Deliverables** (brief description and month of delivery)

 

D.4.1 Demonstration of efficacious prophylactic regimen of best candidate VLP in ferrets. (M21)

Prophylaxis from lethal IAV infection should be 100% after vaccination with the optimal VLP. If this is not the case, as further round of optimisation will be recommended (WP 1.6).

 

D.4.2 Clinical lead candidate selected. (M24)

Reproducible complete protection from lethal challenge will be used as the final go-no-go point for initiating GMP production and, ultimately, clinical trial

 



[1]   Please indicate one activity per work package: RTD = Research and technological development; DEM = Demonstration; MGT = Management of the consortium; OTHER = Other specific activities, if applicable (including any activities to prepare for the dissemination and/or exploitation of project results, and coordination activities).