Description of service

Caracterisation of anti-RSV vaccine candidates: inhibition of viral replication and anti-RSV immune responses, evaluation of vaccine safety.

• We can propose various routes of immunization (intra-nasal, intradermal or subcutaneous) and provide advice on the choice of adjuvant in the case of subunit vaccines.

• We have validated various schedules of immunization: prime followed by one or more boosts, homologous or hererologous prime-boosts.

• We can sample the blood for ELISA assay at various timepoints after immunization.

• For the challenge with RSV, we use a recombinant RSV coding for luciferase (rRSV-luciferase, ref 1) to follow the virus replication in the nasal turbinates and in the lungs of living mice (adults or 6 days old neonates). At different days post-infection, mice are anesthetized and intranasally injected with D-Luciferin. Luminescence emission is evaluated using an in vivo imaging system (IVIS, Emerg’in platform, INRA, Jouy-en-Josas) to determine RSV replication in animals. At day 4, immediately after luminescence reading, mice can also be autopsied in order to perform a and the lungs can be frozen at -80°C until processing for RNA  (qPCR analyses) and protein extractions.

• To monitor inflammation in airways, bronchoalveolar lavage (BAL) is done to isolate BAL cells. Lungs are mashed to release immune cells. Infiltrating BAL and lung cells can then be characterized by FACS analysis or more simply by cytospin and MGG staining.