H5 highly pathogenic avian influenza (HPAI) viruses of the Asian lineage (A/goose/Guangdong/1/96) belonging to clade 2.3.4.4 have spread worldwide through wild bird migration in two major waves: in 2014/2015 (clade 2.3.4.4c), and since 2016 up to now (clade 2.3.4.4b). Due to the increasing risk of these H5 HPAI viruses to establish and persist in the wild bird population, implementing vaccination in certain sensitive areas could be a complementary measure to the disease control strategies already applied. In this study, the efficacy of a novel DNA vaccine, encoding a H5 gene (A/gyrfalcon/Washington/41088-6/2014 strain) of clade 2.3.4.4c was evaluated in specific pathogen-free (SPF) white leghorn chickens against a homologous and heterologous H5 HPAI viruses. A single vaccination at 2 weeks of age (1 dose), and a vaccination at 2 weeks of age, boosted at 4 weeks (2 doses), with or without adjuvant were characterized. The groups that received 1 dose with or without adjuvant as well as 2 doses with adjuvant demonstrated full clinical protection and a significant or complete reduction of viral shedding against homologous challenge at 6 and 25 weeks of age. The heterologous clade 2.3.4.4b challenge of 6-week-old chickens vaccinated with 2 doses with or without adjuvant showed similar results, indicating good cross-protection induced by the DNA vaccine. Long lasting humoral immunity was observed in vaccinated chickens up to 18 or 25 weeks of age, depending on the vaccination schedule. The analysis of viral transmission after homologous challenge showed that sentinels vaccinated with 2 doses with adjuvant were fully protected against mortality with no excretion detected. This study of H5 DNA vaccine efficacy confirmed the important role that this type of so-called third-generation vaccine could play in the fight against H5 HPAI viruses.
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