Immunization and challenge trials in a murine model using different inactivated recombinant vaccines against H1N1 swine influenza virus circulating in Brazil.

Abstract

In Brazil, at least four lineages of influenza A virus circulate pig population: 2009 H1N1 flu pandemic (pH1N1), human-seasonal origin H3N2, H1N1 and H1N2 (huH1 lineages) viruses. Studies related to the occurrence of swine influenza A virus (SIAV) in Brazilian herds have been detecting an increase of occurrence of huH1 lineages. This study aimed to construct recombinant vaccines against the huH1N1 virus and test the immunogens in a murine model. The virus was constructed by reverse genetics using plasmids encoding the HA and NA sequences from a wild huH1N1 virus isolated from an infected pig. Amplified virus was inactivated, and oil-in-water (OW) and gel polymer (GP) adjuvants were used to formulate the vaccines. C57Bl6 mice received two doses with 3 weeks interval by the intramuscular route. Animals were randomly divided into 8 groups (G1-G8): G1 received OW vaccine and G2 PBS plus OW adjuvant; G3 received GP vaccine and G4 PBS plus GP adjuvant; G5 received the live virus by the intranasal route while G6 only PBS; G7 and G8 did not receive any treatment. Serum samples were collected before vaccination and after the first and second dose. Except for G8, three weeks post boost animals were challenged with a wild huH1N1 virus and observed for weight changes. After infection, bronchoalveolar lavage fluid (BALF) and lungs were collected from animals of each group for viral titers and immunohistochemistry (IHC) analysis, respectively. After booster, vaccinated groups seroconverted and the vaccines induced protection upon challenge. Reverse Genetics technique can be used to produce new and quickly updated swine influenza vaccines which is promising to control the virus in Brazilian herds. Future studies may focus on using the technology to produce multivalent recombinant vaccines against distinct strains of SIAVs circulating in Brazilian pig herds.