Data on IL-17 production induced by plant lectins

We reported in article da Silva et al. (2016) [2] that ArtinM induces the IL-17 production through interaction with CD4+ T cells and stimulation of IL-23 and IL-1. Besides ArtinM, other plant lectins (PLs) induce IL-17 production by murine spleen cells. The IL-17 production induced by PLs was evaluated regarding the involvement of IL-23, IL-6, Th1-, and Th2-cytokines. Furthermore, the effect exerted TLR2, TLR4, and CD14 on the PLs׳ performance in the induction of IL-17 was examined. The current data were compared to the known ArtinM ability to induce Th17 immunity.


a b s t r a c t
We reported in article da Silva et al. (2016) [2] that ArtinM induces the IL-17 production through interaction with CD4 þ T cells and stimulation of IL-23 and IL-1. Besides ArtinM, other plant lectins (PLs) induce IL-17 production by murine spleen cells. The IL-17 production induced by PLs was evaluated regarding the involvement of IL-23, IL-6, Th1-, and Th2-cytokines. Furthermore, the effect exerted TLR2, TLR4, and CD14 on the PLs' performance in the induction of IL-17 was examined. The current data were compared to the known ArtinM ability to induce Th17 immunity. &

Value of the data
The report of induction of Th17-immunity by lectins corroborates to the immunomodulatory potential of carbohydrate recognition, opening a new approach to confer protection against pathogens, especially against fungal diseases.
Triggering or maintenance of IL-17 production frequently requires the involvement of cytokines and receptors on immune cells; the identification of such dependence indicates the mechanisms used by a certain lectin to modulate Th17 immunity. This feature is relevant for the choice of which cell population should be used for in vitro studies of immunomodulatory agents.
The data demonstrate that the IL-17 production induced by lectins with distinct sugar-binding specificities instigate further studies with other lectins to analyze the induction of Th17 response, taking this report as an application guidelines.

Data
The data demonstrate that stimulation of murine spleen cells with plant lectins (PL) may induce the IL-17 production, at levels significant higher than the detected in the absence of the stimulus (Tables 1 and 2). Table 1 shows that spleen cells from IL-4 or IFN-γ KO mice under MAL-1 stimulus duplicated the IL-17 production, and the spleen cells from IL-23 KO mice reduced to 66% the levels of IL-17 in response to MAL-1 (Table 1), compared to WT mice. The spleen cells from IL-10 KO mice showed a significant increase in the IL-17 production, when induced by L-PHA, E-PHA, and ConA Table 1 IL-17 production induced by lectins in spleen cells obtained from C57BL/6 (WT) and knockouts for cytokines (KO) mice. The levels of IL-17 in the culture supernatants were measured by ELISA, and the values are expressed as mean 7SEM. One-way ANOVA analysis of variance followed by Bonferroni's post-test was used to compare WT and KO.  Table 2 shows that spleen cells from TLR2 KO, TLR4 KO, and CD14 KO mice not affect significantly the IL-17 production induced by PL.

Experimental design
The suspensions of spleen cells from mice were obtained as described in da Silva et al., [2]. These cells were obtained from all mice strains and used to measure the levels of IL-17 in the culture supernatant. The suspensions of spleen cells (2 Â 10 6 /mL) were cultured in 96-well microplates in the presence of ConA, E-PHA, L-PHA, SNA, MAL, UEA, or Jacalin, all used in final concentrations of 2.5 mg/mL. The reference lectin, ArtinM, was added at concentration of 1.25 mg/mL, and was used phorbol myristate acetate (PMA; Sigma-Aldrich) plus ionomycin (Sigma-Aldrich) as positive control [2]. After 48 h of incubation, the spleen cells were centrifuged (300g, 10 min at room temperature) and the supernatants were collected to measure IL-17A levels. This quantification was determined by an enzyme-linked immunosorbent assay (ELISA) from Ready-SET-Go Kit (eBioscience) according to the manufacturer's instructions. After, the results were analyzed using Prism (Graph Pad Software), and the values are expressed as mean 7standard error of the mean (SEM). Statistical determinations Table 2 IL-17 production induced by lectins in spleen cells obtained from WT and KO for receptors mice. The levels of IL-17 in the culture supernatants were measured by ELISA, and the values are expressed as mean 7 SEM. One-way ANOVA analysis of variance followed by Bonferroni's post-test was used to compare WT and KO. WT  TLR2 KO  TLR4 KO  CD14 KO of the difference between means of groups were performed with analysis of variance (1-way) followed by Bonferroni's multiple comparison tests. * p o0.05 were considered statistically significant.