Mechanistic Insight into the TH1-Biased Immune Response to Recombinant Subunit Vaccines Delivered by Probiotic Bacteria-Derived Outer Membrane Vesicles (original) (raw)

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Figure 4

EcN OMV carriers effectively stimulate adaptive immune cells.

a–c, Representative two-photon microscopy of draining lymph nodes from mice (n = 3, all groups) injected with PBS (a), fluorescently stained EcN OMVs (b), and fluorescently stained EcN OMVs and splenic dendritic cells (DCs) (c). White = collagen, red = OMVs, green = DCs, and yellow = OMV/DC overlap. Arrows denote OMVs (b) or DCs (c). Scale bar = 15 µm. d, Flow cytometry analysis of primary mouse splenic DC stimulation by EcJ and EcN OMVs (n = 3, all groups), identifying subpopulations displaying surface markers for activation CD80 and CD86. e, Proliferation of primary mouse splenic T-cells following coculture with DCs from d (n = 3, all groups) demonstrates more potent activation via EcN OMVs. αCD3/αCD26 = positive control. f, Proliferation of primary mouse bone marrow-derived B-cells following incubation with OMVs, with or without additional T-cell helper factors (n = 3, each group). Anti-IgM/IL-4, anti-CD40/IL-4, and LPS = positive controls. *P<0.01, **P<0.05. All values are given as mean + SD.

Figure 4

doi: https://doi.org/10.1371/journal.pone.0112802.g004