fdOVA encapsulated directly or by the post encapsulation method was released from microparticles, and 1 × 106/mL BM-DCs were incubated overnight with 0.06 μg/mL of released bacteriophages (Fig. 6 ) [10 (link)].
Alternatively, BM-DCs (1 × 106/mL) were left to adhere to multiwell plates. Phage-loaded MPs were resuspended in PBS, and a volume of MPs containing 0.06 μg/mL of encapsulated fdOVA was immediately added to adherent BM-DCs.
BM-DCs were then washed twice and plated at 100,000 cells/well, and the antigen cross presentation was detected by adding the OTI hybridoma cell line B3Z (50,000 cells/well) to the culture. The IL-2 released in the culture medium by activated B3Z cells was measured after 40 h using the supernatants of the co-cultures (0.1 mL/well) and a mouse IL-2 ELISA MAX™ Standard (Biolegend).
Alternatively, BM-DCs (1 × 106/mL) were left to adhere to multiwell plates. Phage-loaded MPs were resuspended in PBS, and a volume of MPs containing 0.06 μg/mL of encapsulated fdOVA was immediately added to adherent BM-DCs.
BM-DCs were then washed twice and plated at 100,000 cells/well, and the antigen cross presentation was detected by adding the OTI hybridoma cell line B3Z (50,000 cells/well) to the culture. The IL-2 released in the culture medium by activated B3Z cells was measured after 40 h using the supernatants of the co-cultures (0.1 mL/well) and a mouse IL-2 ELISA MAX™ Standard (Biolegend).
