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The clinically successful adjuvant MF59 is used in seasonal influenza vaccines, which is proposed to enhance immunity by creating an immune‐competent microenvironment in the muscle that allows recruitment of immune cells that drive adaptive immune responses. Here, we examined whether the clinically successful adjuvants MF59/AddaVax could be used for subcutaneous use and how antigen delivery can be synergized with cellular dynamics at the vaccination site. Subcutaneous injection of AddaVax leads to thickening of the skin, characterized by a neutrophil‐monocyte recruitment sequence. Skin‐infiltrating CCR2 + Ly6C high  monocytes showed differentiation to CD11b + Ly6C + MHCII + CD11c + CD64 +  monocyte‐derived DCs over time in the hypodermal layers of the skin, expressing high levels of CD209a/mDC‐SIGN. Surprisingly, skin thickening was accompanied with increased white adipose tissue highly enriched with monocytes. Analysis of the skin‐draining lymph nodes revealed early increases in neutrophils and moDCs at 12 hours after injection and later increases in migratory cDC2s. Subcutaneous vaccination with AddaVax enhanced antigen‐specific CD8 +  and CD4 +  T cell responses, while moDC targeting using antigen‐coupled CD209a antibody additionally boosted humoral responses. Hence, oil‐in‐water emulsions provide an attractive immune modulatory adjuvants aimed at increasing cellular responses, as well as antibody responses when combined with moDC targeting.

Immunological dynamics after subcutaneous immunization with a squalene‐based oil‐in‐water adjuvant