Characterization of the targeting units within the vaccine proteins.

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Characterization of the targeting units within the vaccine proteins. Characterization of the targeting units within the vaccine proteins. (A) Supernatants from HEK293E cells transiently transfected with the various DNA plasmids were analyzed in sandwich ELISAs as indicated (mean ± SD). (B) Maintenance of CD11c and DEC205 specificity in targeted vaccine proteins. BALB/c splenocytes were stained with supernatants from HEK293E cells transiently transfected with αCD11c-HA or αDEC205-HA plasmids (gray) and gated MHCII+B220− cells analyzed for binding of vaccine proteins. Isotype controls are shown as thick black lines, nonbinding αNIP-HA as a thin line, and mock control as a stippled lined. Bound proteins were detected with mAb that recognize Cγ3. (C) BM-derived Flt-3L DCs from BALB/c mice and the J774A Mφ cell line were analyzed for binding to vaccibody proteins by flow cytometry. DCs were defined as CD45R/B220−CD11c+ cells, and further split into CD11b+ (cDC2) and CD24+ (cDC1) populations, and evaluated for binding to GM-CSF–, Flt-3L–, and FliC-targeted vaccine proteins as compared with nontargeted αNIP-HA vaccines. Ranveig Braathen et al. ImmunoHorizons 2018;2:38-53 Copyright © 2018 The Authors