Variation of Neisseria gonorrhoeae Lipooligosaccharide Directs Dendritic Cell–Induced T Helper Responses
Figure 2
Differential binding of GC glycosylation variants to DCs.
(A) Time course of dose dependent binding of FITC-labeled, fixed GC to immature DCs as analyzed by flow cytometry. GC were incubated for indicated time points at indicated bacteria∶DC ratios. Binding was significantly different for all phenotypes (P<0.05), except for the difference in binding of variants C and D after 15 minutes. (B) Binding and phagocytosis of FITC-labeled, fixed GC to immature DCs was measured by flow cytometry. GC were incubated for 1 hour at indicated bacteria∶DC ratios. Binding and phagocytosis was significantly different for all phenotypes (P<0.05). (C) Phagocytosis of fixed GC (1 hour at indicated bacteria∶DC ratios) can be blocked by the addition of the actin-polymerization inhibitor Cytochalasin D (10 µM). Depicted are the average+SD of triplicates. Cyt D, cytochalasin D (D) Dose dependent binding of FITC-labeled, live GC to immature DCs as analyzed by flow cytometry. GC were incubated for 1.5 hours at indicated bacteria∶DC ratios. (E) Binding of both fixed (left) and live (right) GC phenotypes A and C (1.5 hours at a bacteria∶DC ratio of 20∶1 for fixed and 10∶1 for live GC) to DCs is blocked by 10 mM EGTA. Binding of GC variant C but not A to DCs is inhibited by 100 mM of GalNAc. Depicted are the average+SD of three independent donors for the fixed GC and the average of two independent donors for the live GC. Significant differences: * P<0.05, ** P<0.01, *** P<0.001.
doi: https://doi.org/10.1371/journal.ppat.1000625.g002