Immune-adjuvant activity of lentinan-modified calcium carbonate microparticles on a H₅N₁ vaccine

In recent years, the high prevalence of avian influenza viruses especially H₅N₁ subtype isolated from poultry and human has become a major public health concern. Vaccination is still a major strategy for preventing H₅N₁ infections. Lentinan (LNT), a β-1,3-glucohexaose with β-1,6-branches, is extracted from Lentinus edodes and has been extensively studied for its immunoenhancement effects. In this study, we synthesized and characterized calcium carbonate (CaCO₃) microparticles which modified with LNT as an adjuvant for H₅N₁ vaccine and investigated their ability to enhance immune responses. We prepared spherical and uniform CaCO₃-LNT microparticles with a mean hydrodynamic size was around 2 μm. The H₅N₁ antigen-loaded CaCO₃-LNT particles were injected into mice to evaluate their effectiveness as an adjuvant for H₅N₁ vaccines. The results demonstrated that CaCO₃-LNT/H₅N₁ significantly enhanced the expression of MHC-II and CD86 in lymph node dendritic cells, and increased the ratio of CD4⁺ to CD8⁺ T cells in lymphocytes. Moreover, CaCO₃-LNT/H₅N₁ surprisingly increased the HI titers and induced the secretion of IgG subtypes (IgG1 and IgG2b) and Th-associated cytokines (TNF-α, IFN-γ and IL-4) in immunized mice. Therefore, by combining with the immunostimulatory activity of LNT and the drug/antigen delivery capabilities of CaCO₃, the CaCO₃-LNT/H₅N₁ could induce a stronger cellular and humoral immune response and could be a potential adjuvant for the H₅N₁ vaccine.