Green preparation of hydrogel particles-in-emulsions for simultaneous enhancement of humoral and cell-mediated immunity

Emulsions are one of the most often used vaccine adjuvant formulations. Although they promote high humoral immunity, the induced cellular immunity is often poor, which restrict their application. To enhance the cellular immunity, some researchers have prepared mixed formulations by adding particles into the aqueous phase of emulsions. However, the particle preparation process usually involves the addition and removal of organic reagents, which is environmentally unfriendly and cumbersome. Moreover, the obtained vaccine adjuvant only induces limited cell-mediated immunity and humoral immunity compared with emulsion-adjuvanted vaccines. Herein, we developed a green and simple method for fabricating a novel nanoparticles-in-emulsions (NPE) formulation. Firstly, a temperature-sensitive hydrogel was used to prepare particles by self-solidification without additional crosslinking reagents. Secondly, the white oil was used as organic phase to avoid the particle washing procedures and organic solvent residues. Moreover, the effect of NPE as vaccine adjuvant was evaluated by using two veterinary vaccines as model antigens. NPE showed advantages than the conventional vaccine formulations in inducing both humoral and cellular immunity. This work provides a facile and broadly applicable approach for preparing nanoparticles-in-emulsions formulation, and presents an effective adjuvant for enhancing immunity against infectious diseases.