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Adjuvants enhance vaccine immunogenicity but their mechanism of action is often incompletely understood, hampering rapid applicability for pandemic vaccines. Herein, we characterized the cellular and molecular activity of several adjuvant formulations available for pre-clinical evaluation, including several developed for global open access. We applied four complementary human in vitro platforms to assess individual and combined adjuvants in soluble, oil-in-water, and liposomal formulations. Of all formulations tested, liposomal co-formulation of MPL and QS-21 was most potent in promoting dendritic cell maturation, selective production of Th1-polarizing cytokines, differentiation of mature dendritic cells in a microphysiological 3D tissue construct, and activation of antigen-specific CD4+ and CD8+ T cells in a SARS-CoV-2 spike antigen-specific co-culture assay. Thus, human in vitro modeling provides insight into …