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. Author manuscript; available in PMC: 2015 Jun 1.
Published in final edited form as: Nat Med. 2014 Nov 10;20(12):1458–1463. doi: 10.1038/nm.3709

Figure 4. Murine and human lupus sera stimulate dermal dendritic cell migration to lymph nodes in vivo.

Figure 4

(a) Cell tracks (white) of footpad dermal DCs (green) 16 hours following the administration of heat-inactivated, aged WT mouse serum, or serum obtained from autoimmune NZB/W F1 mice. Blood vessels shown in red (Qdots IV) and cell tracks over an 80 minute period in white. Scale bar = 50μm. Quantification of displacement and persistence index of dermal DCs from three mice in each group is shown in (b). (c) Quantification of displacement and persistence index of footpad dermal DCs in CD11cEYFP mice 16 hours following the administration of heat-inactivated, NZB/W F1 mouse serum containing IgG (IgG+) or an aliquot depleted of IgG (IgG-). (d) Number of live/FITC+/MHCIIhigh/CD11c+ cells per draining lymph node in WT mice and (e) Representative flow cytometric plots (left panel) and quantification (right panel) of FITC+/MHCIIhigh/CD11c+/EPCAM+ (Langerhans cells) and CD11c+/CD103+ (dermal DCs) in draining lymph nodes 48 hours after application of FITC to skin in mice given heat-inactivated WT (open circles) or SLE (black circles) serum intraperitoneally 12 hours previously. Each circle in d and e represents quantification from a single lymph node. (f) Number of CD11c+/MHCIIhigh/CD103+ cells in renal lymph nodes of aged NZM mice with lupus nephritis and aged-match NZW controls. Graphs show mean and SEM of lymph nodes taken from four mice. (g) Representative cell tracks (white) of footpad dermal DCs (green) and (h) Quantification of displacement and persistence index of dermal DCs in WT mice 16 hours following the administration of heat-inactivated serum obtained from healthy controls or from patients with SLE. Scale bar in g = 50 μm. Graphs show values from mice treated with three healthy control sera (open circles) or from three SLE patients with low disease activity (BILAG scores 2-9, filled squares, ‘SLE low’) or three SLE patients with high disease activity (BILAG scores 15-21, open squares, ‘SLE high’). (i) CCR7 expression on WT U937 cells or those stably expressing FcγRIIB-I232 or FcγRIIB-T232 24 hours following incubation with OVA or IC. Graph shows mean and SEM of duplicates from one of three experiments. In (b), (c), and (h) each point represents a single dermal DC, and the bar shows the mean value. P values calculated using a Students t test (two-tailed). * = P ≤ 0.05, ** = P ≤ 0.01, *** = P ≤ 0.001, **** = P ≤ 0.0001.