Figure 4
Figure 4. FcγRs on human BDCA-3+ contribute to antigen cross-presentation. FcγRI, II and III expression on BDCA-1+ and BDCA-3+ DCs extracted from human blood (A) and tonsils (B; BDCA-1+, dashed line) and (BDCA-3+, solid line; n = 3). (C) FcγRIIa (black bars) and FcγRIIb (white bars) mRNA expression in BDCA-1+ and BDCA-3+ DCs extracted from human blood (mean + SEM, n = 5). (D) relative FcγRIIa (black bars) and FcγRIIb (white bars) mRNA expression in BDCA-1+ and BDCA-3+ DCs (mean + SEM, n = 5). (E) BDCA-1+ (white bar) and BDCA-3+ (black bar) were FACS-sorted (at least 98% purity) and cultured in the presence of 3 μg pp65 (left bars) or pp65-IC (right bars; O/N, 37°C). Next, A2/NLVPMVATVCD8+ T cells were added to DC cultures and cross-presentation was analyzed as in Figures 1 and 2 (mean + SEM, n = 3). (F-G) Representative plots and summary of IFN-γ production (mean + SEM) after IC-mediated cross-presentation in the absence (black bar) or presence of FcR blocking reagents (IgG-Fc fragments (light gray), recombinant S aureus–FLIPr-like (dark gray; n = 3 in panel F; n = 2 in panel G).

FcγRs on human BDCA-3+ contribute to antigen cross-presentation. FcγRI, II and III expression on BDCA-1+ and BDCA-3+ DCs extracted from human blood (A) and tonsils (B; BDCA-1+, dashed line) and (BDCA-3+, solid line; n = 3). (C) FcγRIIa (black bars) and FcγRIIb (white bars) mRNA expression in BDCA-1+ and BDCA-3+ DCs extracted from human blood (mean + SEM, n = 5). (D) relative FcγRIIa (black bars) and FcγRIIb (white bars) mRNA expression in BDCA-1+ and BDCA-3+ DCs (mean + SEM, n = 5). (E) BDCA-1+ (white bar) and BDCA-3+ (black bar) were FACS-sorted (at least 98% purity) and cultured in the presence of 3 μg pp65 (left bars) or pp65-IC (right bars; O/N, 37°C). Next, A2/NLVPMVATVCD8+ T cells were added to DC cultures and cross-presentation was analyzed as in Figures 1 and 2 (mean + SEM, n = 3). (F-G) Representative plots and summary of IFN-γ production (mean + SEM) after IC-mediated cross-presentation in the absence (black bar) or presence of FcR blocking reagents (IgG-Fc fragments (light gray), recombinant S aureus–FLIPr-like (dark gray; n = 3 in panel F; n = 2 in panel G).

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