The immunogenicity-enhancing effect of emulsion vaccine adjuvants is independent of the dispersion type and antigen release rate--a revisit of the role of the hydrophile-lipophile balance (HLB) value.

Effective antigen delivery is one of the most important issues in vaccine development. It has been suggested that adjuvant action results from a depot effect by prolonging the duration of the interaction between antigen and cells, and thus is related to the antigen-releasing properties of emulsion adjuvants. The objective of this study was to investigate the effect of the dispersion properties of emulsion-type vaccine adjuvants on the immune response with the aim of optimizing vaccine adjuvant formulation. Emulsion-type adjuvants with various dispersion properties of either the oil-in-water or water-in-oil type were prepared using emulsifiers with various hydrophilic-hydrophobic balance (HLB) values. The physicochemical properties of the emulsions, including the conductivity and viscosity, and antigen release rates were then determined. Cell death induced by the vaccine adjuvants was examined in EL4 cells by Annexin V/propidium iodide (PI) staining and flow cytometric analysis. Mice were immunized with or without the adjuvants and the immunogenicity-enhancing effect of the adjuvants determined by measuring antibody production using an enzyme linked immunosorbent assay. The conductivity, viscosity, and antigen release rates varied widely among emulsions containing emulsifiers with different HLB values. However, the magnitude of the antigen-specific antibody response was similar in most emulsions adjuvants containing Spans or Tweens. L121-adjuvant, the control adjuvant inducing the strongest apoptosis in vitro, was shown to stimulate the highest antibody response in vivo. The results obtained in this study indicate that the immunogenicity-enhancing effect of emulsion adjuvants is independent of the dispersion type and the antigen release rate of the vaccine delivery system.