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We encapsulated tumor necrosis factor-α (TNF-α), a major proinflammatory cytokine, into cholesteryl pullulan (CHP) to prepare TNF/CHP nanoparticles. In this chapter, the immune response-enhancing capability of the nanoparticles to act as a vaccine adjuvant against influenza is described. TNF/CHP nanoparticles showed excellent storage stability, and they enhanced host immune responses to external immunogens. We applied the nanoparticles in a mouse model of influenza virus infection to investigate their adjuvant ability. Nasal administration of TNF/CHP nanoparticles combined with a conventional split vaccine was effective at inducing systemic IgG1 as well as mucosal IgA, and it protected mice against a lethal challenge of A/PR/8/34 (H1N1) influenza virus. Mechanistic studies showed that the nanoparticles enhanced antigen uptake by dendritic cells (DCs) and moderately induced the expression of inflammation-related genes in nasal-associated lymphoid tissue (NALT), leading to the activation of both B and T cells. A preliminary safety study revealed no severe toxicity to TNF/CHP nanoparticles. Slight-to-moderate influences in nasal mucosa were observed only after repeated administration and they were reversible. Our data show that TNF/CHP nanoparticles effectively enhance both humoral and cellular immunity via nasal administration and could be a potential adjuvant for vaccines against infectious diseases like influenza.

Cholesteryl Pullulan Nanoparticles-Encapsulated TNF-α: An Effective Mucosal Vaccine Adjuvant Against Influenza