M2e conjugated gold nanoparticle influenza vaccine displays thermal stability at elevated temperatures and confers protection to ferrets.

Abstract

Currently approved influenza vaccines are not only limited in breadth of protection but also have a limited shelf-life of 12-18\xa0months when stored under appropriate conditions (2-8\xa0°C). Inadvertent alteration in storage temperatures during manufacturing, transportation, distribution until delivery to patient, can damage the vaccine thus reducing its efficacy. A thermally stable vaccine can decrease the economic burden by reducing reliance on refrigeration system and can also enhance outreach of the vaccination program by allowing transportation to remote areas of the world where refrigerated conditions are scarce. We have previously developed a broadly protective influenza A vaccine by coupling the highly conserved extracellular region of the matrix 2 protein (M2e) of influenza A virus to gold nanoparticles (AuNPs) and upon subsequent addition of toll-like receptor 9 agonist - CpG, as an adjuvant, have shown its breadth of protection in a mouse model. In this study, we show that the vaccine is thermally stable when stored at 4\xa0°C for 3\xa0months, 37\xa0°C for 3\xa0months and 50\xa0°C for 2\xa0weeks in its lyophilized form, and later it was possible to readily reconstitute it in water without aggregation. Intranasal vaccination of mice using reconstituted vaccine induced M2e-specific IgG and IgG subtypes in serum similar to the freshly formulated vaccine, and fully protected mice against lethal influenza A challenge. Immunization of ferrets intranasally or intramuscularly with the vaccine induced M2e-specific IgG and there was reduced virus level in nasal wash of ferrets immunized through intranasal route.