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. 2017 Jul 27;12(7):e0181866. doi: 10.1371/journal.pone.0181866

Fig 6. CBirTox induces IgA production from naïve B cells in vitro.

Fig 6

Splenic CD11c+ DCs were pulsed with 1 μg/ml CBirTox or untreated for 4 hours before co-culture with CD19+ PP B cells (A). Supernatants were collected after 7 days and examined for IgA production via ELISA. Results are expressed as the mean ± SEM of 3–6 independent experiments. Groups were analyzed using student’s unpaired t test. (B) Splenic CD11c+ DCs were pulsed with 1 μg/ml CBirTox, 1 μg/ml CTB, 1 μg/ml CTB plus 1 μg/ml CBir1 peptide,1 μg/ml CBir1 carboxy-domain or untreated for 4 hours before co-culture with CD43- splenic B cells at a 1:5 ratio. Supernatants were collected after 7 days and examined for IgA production via ELISA. Results are expressed as the mean ± SEM of 3 independent experiments. Groups were analyzed using one-way ANOVA with Tukey’s post test. (C) Splenic CD11c+ DCs were pulsed with 1 μg/ml CBirTox or untreated for 4 hours before co-culture with CD43- splenic B cells at a 1:5 ratio with or without the addition of RA inhibitor LE135 or anti-TGF-β receptor I kinase III at 1 μM. Supernatants were collected after 7 days and examined for IgA production via ELISA. Results are expressed as the mean ± SEM of 4 independent experiments. Groups were analyzed using one-way ANOVA with Kruskal-Wallis correction. *p<0.05, **p<0.005, ***p<0.0005, NS, not significant.