Immature lung TNFR2(−) conventional DC 2 subpopulation activates moDCs to promote cyclic di-GMP mucosal adjuvant responses in vivo

Cyclic dinucleotides (CDNs), including cyclic di-GMP (CDG), are promising vaccine adjuvants in pre-clinical/clinical trials. The in vivo mechanisms of CDNs is not clear. Here we investigated the roles of lung DCs subsets in promoting CDG mucosal adjuvant responses in vivo. Using genetically modified mice and adoptive cell transfer, we identified lung conventional DC 2 (cDC2) as the central player in CDG mucosal responses. We further identified two functionally distinct lung cDC2 subpopulations: TNFR2(+)pRelB(+) and TNFR2(−)pRelB(−) cDC2. The TNFR2(+) cDC2 were mature and migratory upon intranasal CDG administration while the TNFR2(−) cDC2 were activated but not mature. Adoptive cell transfer showed that TNFR2(−) cDC2 mediate the antibody responses of CDG, while the TNFR2(+) cDC2 generate Th1/17 responses. Mechanistically, immature TNFR2(−) cDC2 activate monocyte-derived DCs (moDCs), which do not take up intranasally administered CDG. moDCs promote CDG-induced generation of T follicular helper- and germinal center B-cells in the lungs. Our data revealed a previously undescribed in vivo mode of DCs action whereby an immature lung TNFR2(−) cDC2 subpopulation directs the non-migratory moDCs to generate CDG mucosal responses in the lung.