Evaluating dendritic cells as an in vitro screening tool for immunotherapeutic formulations

Abstract

Immunotherapy approaches targeting dendritic cells (DCs) are being studied as treatment options in cancer. This project focused on utilizing DCs as a valuable in vitro screening tool for efficacious microparticle formulations containing tumor associated antigens (TAAs) and adjuvants as immunotherapy alternatives. The innate immune system, including DCs, distinctly responds to the particulate matter and adjuvants in these formulations which stimulates the adaptive immune system to eliminate resident cancer cells. We formulated microparticles (MPs) co-loaded with TAAs along with the adjuvants, AddaVax™ and Imiquimod, and measured their effect on DCs in eliciting a cell-mediated immune response towards tumors. The MP zeta potential was measured as −24.0 mV and −26.5 mV for blank and TAA/adjuvant co-loaded microparticles, and the average particle size was 671.2 nm and 854.4 nm respectively. We determined that nitric oxide (NO) secretion was significantly higher in the adjuvant MP treated DCs group and was dose dependent with 1 mg/mL demonstrating the highest secretion levels. TNF-α release was highest in AddaVax™/TAA and Imiquimod/TAA MPs treated DCs, while IL-6 secretion was highest from Imiquimod/TAA MPs as well as from combined AddaVax™/TAA and Imiquimod/TAA MPs. Overall, the cell surface marker expressions of CD80, CD86, CD40, CD54, MHC-I and MHC-II levels were highest with combined AddaVax™/TAA and Imiquimod/TAA MPs. The results of our experiments suggest that a combination of adjuvants targeting different DC receptors loaded with TAA MPs creates an efficient delivery system to T-cells that could improve adaptive immune responses. Our studies also confirm that DCs are potent innate immune cells that can be used successfully as an in vitro tool to screen novel delivery formulations focused on immunotherapy.