Improved immune responses in mice using the novel chitosan adjuvant ViscoGel, with a Haemophilus influenzae type b glycoconjugate vaccine

Abstract

An immune response to an antigen is more efficiently induced in combination with an adjuvant. Chitosan has due to documented immunostimulatory characteristics been proposed as an adjuvant candidate. However, a disadvantage with chitosan is its poor solubility at physiological pH. We have circumvented this obstacle by using a soluble type of chitosan (Viscosan), with a degree of deacetylation (DD) of 50% and a random distribution of acetyl groups. A hydrogel, ViscoGel, was made from Viscosan which was further mechanically processed into gel particles of predefined size. The first cells to infiltrate ViscoGel in mice, were identified mainly as neutrophils, detected already after 4 h. ViscoGel's impact on the immune response in mice together with a commercial vaccine against Haemophilus influenzae type b (Act-HIB) was then studied. Mixing Act-HIB with ViscoGel, induced significantly enhanced IgG1 and IgG2a titers in serum (p<0.05). We could reduce the antigen dose ten-fold in combination with ViscoGel and still obtain antibody titers similar to 2 μg Act-HIB administered alone. In addition, the Act-HIB specific cellular response was stronger in mice vaccinated together with ViscoGel (p<0.05). The cytokine response after vaccination with Act-Hib together with ViscoGel was of a mixed type. We found elevated levels of the Th1 associated cytokine INF-γ, the Th2-cytokine IL-4, the proinflammatory IL-6 and IL-17A, and the regulatory cytokine IL-10. Similar effects were seen when the adjuvant was administered either subcutaneously or intramuscularly. Taken together, using vaccination against H. influenzae type b as a model, we here show proof of concept for the novel vaccine adjuvant candidate, ViscoGel.