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B cell ELISPOT assay for DENV-specific antibody-secreting cells. NHP PBMCs were stimulated ex vivo with IL-2 and R848 for 5 days and then incubated with 5 μg/ml of DENV1 to -4 E proteins, anti-monkey IgG, or BSA. The total number of memory B cells (measured by their conversion to IgG-producing cells) was counted for each DENV E protein or BSA (x axis) in preimmune (a) and naive (b) NHP vaccination groups.
