Evaluating -galactosylceramide as an adjuvant for live attenuated influenza vaccines in pigs

Natural killer T (NKT) cells activated with the glycolipid ligand alpha-galactosylceramide (alpha-GalCer) stimulate a wide variety of immune cells that enhance vaccine-mediated immune responses. Several studies have used this approach to adjuvant inactivated and subunit influenza A virus (IAV) vaccines, including to enhance cross-protective influenza immunity. However, less is known about whether alpha-GalCer can enhance live attenuated influenza virus (LAIV) vaccines, which usually induce superior heterologous and heterosubtypic immunity compared to non-replicating influenza vaccines. The current study used the swine influenza challenge model to assess whether alpha-GalCer can enhance cross-protective immune responses elicited by a recombinant H3N2 LAIV vaccine (TX98 Delta NS1) encoding a truncated NS1 protein. In one study, weaning pigs were administered the H3N2 TX98 Delta NS1 LAIV vaccine with 0, 10, 50, and 100 mu g/kg doses of alpha-GalCer, and subsequently challenged with a heterologous H3N2 virus. All treatment groups were protected from infection. However, the addition of alpha-GalCer appeared to suppress nasal shedding of the LAIV vaccine. In another experiment, pigs vaccinated with the H3N2 LAIV, with or without 50 mu g/kg of alpha-GalCer, were challenged with the heterosubtypic pandemic H1N1 virus. Pigs vaccinated with the LAIV alone generated cross-reactive humoral and cellular responses which blocked virus replication in the airways, and significantly decreased virus shedding. On the other hand, combining the vaccine with alpha-GalCer reduced cross-protective cellular and antibody responses, and resulted in higher virus titers in respiratory tissues. These findings suggest that: (i) high doses of alpha-GalCer impair the replication and nasal shedding of the LAIV vaccine; and (ii) alpha-GalCer might interfere with heterosubtypic cross-protective immune responses. This research raise concerns that should be considered before trying to use NKT cell agonists as a possible adjuvant approach for LAIV vaccines.