Fig 1: IgA-coated BEVs promote pro-inflammatory cytokine release in CD89+ cells.a IL-8 response in CD89+CD14+CD11b+ U937 monocytes exposed to uncoated or IgA-coated BEVs from B. fragilis and L. acidophilus (BEVsB/L and IgA-BEVsB/L), uncoated or IgA-coated hEVs or soluble IgA alone. Dosage of the effector samples given by total protein biomass determined by Bradford is indicated. Incubation with saline served as mock-treated control (co). (n = 10 for each data set). b IL-6 response in CD89+CD14+CD11b+ U937 monocytes exposed to uncoated or IgA-coated BEVs from B. fragilis and L. acidophilus (BEVsB/L and IgA-BEVsB/L), uncoated or IgA-coated hEVs or soluble IgA alone. Dosage of the effector samples given a total protein biomass determined Bradford is indicated. Incubation with saline served as mock-treated control (co). (n = 8 for uncoated hEVs; all other data sets n = 9). c IL-8 response in CD89-negative human intestinal organoid-derived monolayers from two healthy donors exposed to 5 µg uncoated or IgA-coated BEVs from B. fragilis and L. acidophilus (BEVsB/L and IgA-BEVsB/L). Lack of CD89 expression was confirmed by FACS (Fig. 3a). Data is presented as median ± interquartile range (n = 5 for each data set). d IL-8 (left) and IL-6 (right) response in CD14+ human monocytes isolated from three healthy donors exposed to 5 µg uncoated or IgA-coated BEVs from B. fragilis and L. acidophilus (BEVsB/L and IgA-BEVsB/L). CD89 expression was confirmed by FACS analysis (Fig. 3b). Data is presented as median ± interquartile range (n = 4 for donor 1, n = 5 for donor 2, and n = 7 for donor 3). a–d Data is presented as median ± interquartile range. Samples with no detectable signal were set to the limit of detection, which is indicated by a dotted line. Statistical difference between uncoated and IgA-coated BEVs was evaluated by two-sided Mann–Whitney U-test. Source data are provided as a Source Data file.
Fig 2: Identification of a pro-inflammatory property in EV fractions derived from UC patients.a IL-8 response in HT-29 human intestinal epithelial cells exposed to soluble effector (SEF, left) and extracellular vesicle (EV, right) fractions of the non-IBD (blue), inactive UC (yellow) and active UC (red) group. b IgA titers in soluble effector (SEF, left) and extracellular vesicle (EV, right) fractions of the non-IBD (blue), inactive UC (yellow), and active UC (red) group. c IgG titers in soluble effector (SEF, left) and extracellular vesicle (EV, right) of the non-IBD (blue), inactive UC (yellow), and active UC (red) group. d sCD89-IgA complex levels in soluble effector (SEF, left) and extracellular vesicle (EV, right) fractions of the non-IBD (blue), inactive UC (yellow), and active UC (red) group. e Representative images showing immunofluorescent stainings of human colonic biopsies derived from a non-IBD, inactive UC, and active UC patient for CD89 (red), immune cell marker CD68 (green) and total nuclei stained with DAPI (blue). Graph on the right side depicts the number of CD89+CD68+ and CD89+CD68dim/- cells per eye field for independent biopsies (non-IBD, n = 9; inactive UC, n = 10; active UC, n = 12). Scale bar = 50 µm. Data in graphs is presented as median ± interquartile range. Statistical difference against the non-IBD group was evaluated by Kruskal–Wallis with Dunn’s test. f IL-8 response in CD89+CD14+CD11b+ U937 monocytes exposed to soluble effector (SEF, left) and extracellular vesicle (EV, right) fractions of the non-IBD (blue), inactive UC (yellow) and active UC (red) group. g IL-6 response in the in CD89+CD14+CD11b+ U937 monocytes exposed to soluble effector (SEF, left) and extracellular vesicle (EV, right) of the non-IBD (blue), inactive UC (yellow) and active UC (red) group. a–d, f, g sample number as follows: non-IBD (n = 28), inactive UC (n = 30) and active UC (n = 16). Data is presented as median ± interquartile range. Samples with no detectable signal were set to the limit of detection, which is indicated by a dotted line. Statistical difference between the three groups was evaluated by Kruskal–Wallis with Dunn’s test. a–g Source data are provided as a Source Data file.
Fig 3: IgA-coated BEVs and EV fractions from UC patients promote a CD89-dependent pro-inflammatory cytokine release in mouse bone marrow derived cells (BMDC).a IL-6 and TNF-α response in bone marrow derived cells (BMDC) from CD89wt/wt (littermates, LM) and CD89tg/wt (CD89-transgenic, CD89) mice exposed to uncoated or IgA-coated BEVs from B. fragilis and L. acidophilus (BEVsB/L and IgA-BEVsB/L) as well as soluble IgA alone. Dosage of the effector samples given by total protein biomass determined by Bradford is indicated. Incubation with saline served as mock-treated control (co). (n = 10 for each data set). b IL-6 and TNF-α response in bone marrow derived cells (BMDC) from CD89tg/wt (CD89-transgenic, CD89) mice exposed to 1 µg IgA-coated BEVs from B. fragilis and L. acidophilus (IgA-BEVsB/L) in the absence (-) or presence (+) of the Syk inhibitor R406. (n = 6 for each data set). c IL-6 (left) and TNF-α (right) response in bone marrow derived cells (BMDC) from CD89wt/wt (littermates, LM) and CD89tg/wt (CD89-transgenic, CD89) mice exposed to EV fractions derived from the non-IBD (blue) and active UC (red) group (seven randomly chosen representatives). (n = 7 for each data set). d IL-6 and TNF-α response in bone marrow derived cells (BMDC) from CD89tg/wt (CD89-transgenic, CD89) mice exposed to 1 µg EV fractions derived from three representative active UC patients (UC1, UC2, or UC3) in the absence (-) or presence (+) of the Syk inhibitor R406. (n = 4 for each data set). a–d Data is presented as median ± interquartile range. Samples with no detectable signal were set to the limit of detection, which is indicated by a dotted line. Statistical difference in panel a was evaluated by Kruskal–Wallis (comparison of groups receiving the same BEVsB/L dosage) with Dunn’s test. Statistical differences in all other panels [comparison of (B)EV samples with and without inhibitor or non-IBD and active UC groups] were evaluated by two-sided Mann–Whitney U-test. Source data are provided as a Source Data file.
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