Dendritic cell activation by a micro particulate based system containing the influenza matrix-2 protein virus-like particle (M2e VLP)

M2e VLP was previously described as a vaccine that incorporates the extracellular region of the matrix 2 protein (M2e), which is highly conserved amongst all the strains of influenza. In this study, we analyzed activation status of dendritic cells (DCs) after exposure to M2e VLP, stimulating DCs with M2e VLP and co-culturing the stimulated DCs with T cells to observe innate and adaptive immune responses. The M2e VLP microparticle was prepared by encapsulating into a polymer matrix using the one-step spray drying method. Adjuvants Alhydrogel (R), MPL-A (R) or Addavax (TM) were used to enhance the DC stimulatory effects by the M2e VLP microparticle. The M2e VLP microparticle yield was found to be 92% and the encapsulation yield was around 84% with a size of approximately 2.78 mu m. There was no short-term cytotoxicity found in DCs and macrophages with concentrations up to 1500 mu g/mL of M2e VLP microparticle, however long-term exposure resulted in 25% decrease in viability of cells with concentrations more than or equal to 500 mu g/mL. The M2e VLP microparticle vaccine with Alhydrogel (R) and MPL-A (R) induced high levels of TNFix in both DCs and macrophages. The high levels of MHC I, II, CD28, B7-1, ICAM-1, LFA-1 expression and IL-12 release in the M2e VLP microparticle group with Alhydrogel (R) suggests that the M2e VLP vaccine with this adjuvant activated T cells via the Th2 pathway. The increased expression of MHC I, II, CD40, CD154, ICAM-1 and LFA-1 on DCs and the release of IL-12 in the M2e VLP microparticle culture of DCs with MPL-A (R) demonstrated that the M2e VLP vaccine with this adjuvant activated T cells via the Th1 pathway. The decrease in fluorescence in the Alhydrogel (R) and MPL-A (R) group illustrates the proliferation of T cells took place following exposure of DCs to the M2e VLP microparticle with these adjuvants. The M2e VLP microparticle exhibited higher stimulatory responses of DCs than the M2e VLP in suspension. Furthermore, the presence of Alhydrogel (R) and MPL-A (R) enhanced the stimulatory effects of DCs by the M2e VLP microparticle (MP) vaccine.

Automated summary

This study examined the activation status of dendritic cells (DCs) and the innate and adaptive immune responses after exposure to M2e VLP vaccine. Results showed that the M2e VLP microparticle had a strong stimulatory effect on DCs, and the presence of Alhydrogel(R) and MPL-A(R) adjuvants enhanced this effect.