Influence of the administration route and dose on the expression and antibody responses of a reporter and avian influenza self-amplifying mRNA vaccine in poultry.

Vaccination is routinely used in industrial poultry to control infectious diseases. Vaccines based on mRNA and self-amplifying RNA (saRNA) are approved for human use, but research on their application in poultry is limited. In this study the saRNA vaccine platform is evaluated in poultry. First, a luciferase-encoding saRNA (luc-saRNA) was tested as a model vaccine across different administration routes and doses in broilers. High luciferase expression, and anti-luciferase antibodies were observed after intramuscular (IM), subcutaneous (SC), and in ovo (IO) administration. After a second Luc-saRNA injection, seroconversion rates and antibody titers increased in the IM and SC group to almost 100%. Higher doses of Luc-saRNA increased luciferase production. However, they did not linearly increase antibody production, as all tested doses (0.20-5.0 µg) elicited an equipotent immune response. A vaccination experiment with saRNA encoding the hemagglutinin head-domain (HA-HD) of H5N1 avian influenza showed hemagglutinin inhibition (HI) titers that are indicative for protection after a single injection and these titers remained above the protective threshold during 6 weeks without boosting. When boosted, the HI titers increased four-fold. This study confirms effective protein translation and immune response induction in chickens with IM or SC administered saRNA-LNPs, even at the lowest dose of 0.20 µg.