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Ascaris lumbricoides Cystatin Prevents Development of Allergic Airway Inflammation in a Mouse Model

Coronado, Sandra, Zakzuk, Josefina, Regino, Ronald, Ahumada, Velky, Benedetti, Ines, Angelina, Alba, Palomares, Oscar, Caraballo, Luis·2019·Frontiers in Immunology
DOI 10.3389/fimmu.2019.02280PMC PMC6777510
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Figure 1 Influence of rAl-CPI on allergic airway hyper-reactivity and inflammatory infiltrate. (A) Scheme of the experimental protocol used in the animal model of allergic airways inflammation. (B) MCh induced airway reactivity was indirectly evaluated by whole body plethysmography. (C) Eosinophil and neutrophil numbers in BAL. (D) Total inflammation scores obtained from the histopathological analyses in lung tissue sections from each experimental group. (E) Representative photomicrographs (X100) from the different groups are shown. Areas with PV or PB inflammation are labeled in H/E stained slides (top row). (F) PAS-positive cells are observed in magenta (indicated by arrowheads). Mean values ± SEM are shown for 6–7 mice per group; representative data of three independent experiments. One way ANOVA + Dunnet, comparison with PBS and BtE controls, two-way ANOVA for methacholine concentration, Kruskall Wallis for inflammation score and Goblet cells. *p ≤ 0.05; ***p ≤ 0.001.
Figure 1 Influence of rAl-CPI on allergic airway hyper-reactivity and inflammatory infiltrate. (A) Scheme of the experimental protocol used in the animal model of allergic airways inflammation. (B) MCh induced airway reactivity was indirectly evaluated by whole body plethysmography. (C) Eosinophil and neutrophil numbers in BAL. (D) Total inflammation scores obtained from the histopathological analyses in lung tissue sections from each experimental group. (E) Representative photomicrographs (X100) from the different groups are shown. Areas with PV or PB inflammation are labeled in H/E stained slides (top row). (F) PAS-positive cells are observed in magenta (indicated by arrowheads). Mean values ± SEM are shown for 6–7 mice per group; representative data of three independent experiments. One way ANOVA + Dunnet, comparison with PBS and BtE controls, two-way ANOVA for methacholine concentration, Kruskall Wallis for inflammation score and Goblet cells. *p ≤ 0.05; ***p ≤ 0.001.CC BY · PubMed Central
Figure 2 rAl-CPI interferes with IgE production and potentiates IgG2a response. (A) Serum Total IgE, IgG, IgG1, and IgG2a antibody levels. (B) Serum BtE allergen-specific IgE, IgG IgG1, and IgG2a antibody levels. The mean values ± SEM are shown for 6–7 mice per group; representative data from three independent experiments. One-way ANOVA with Dunnet post-hoc analysis). ***p ≤ 0.001.
Figure 2 rAl-CPI interferes with IgE production and potentiates IgG2a response. (A) Serum Total IgE, IgG, IgG1, and IgG2a antibody levels. (B) Serum BtE allergen-specific IgE, IgG IgG1, and IgG2a antibody levels. The mean values ± SEM are shown for 6–7 mice per group; representative data from three independent experiments. One-way ANOVA with Dunnet post-hoc analysis). ***p ≤ 0.001.CC BY · PubMed Central
Figure 3 rAl-CPI modifies local and systemic cytokine responses. (A) Th2 cytokines detected in BAL: IL-5, IL-13, and IL-10. (B) Cytokines in spleen cell cultures restimulated in vitro with BtE (for group rAl-CPI culture was stimulated with rAl-CPI), IL-4, IL5, IL-13, IFNγ, IL-10, and LAP-TGFβ. The mean values ± SEM are shown for 6–7 mice per group; representative data from three independent experiments. One-way ANOVA with Dunnet post-hoc analysis. *p ≤ 0.05; ***p ≤ 0.001.
Figure 3 rAl-CPI modifies local and systemic cytokine responses. (A) Th2 cytokines detected in BAL: IL-5, IL-13, and IL-10. (B) Cytokines in spleen cell cultures restimulated in vitro with BtE (for group rAl-CPI culture was stimulated with rAl-CPI), IL-4, IL5, IL-13, IFNγ, IL-10, and LAP-TGFβ. The mean values ± SEM are shown for 6–7 mice per group; representative data from three independent experiments. One-way ANOVA with Dunnet post-hoc analysis. *p ≤ 0.05; ***p ≤ 0.001.CC BY · PubMed Central
Figure 4 rAl-CPI stimulates Treg differentiation. (A) Percentage of Treg cells in spleen. (B) Percentage of Treg cells when the anti-IL-10R monoclonal or isotype control was used. The mean values ± SEM are shown for 6–7 mice per group; representative data from three independent experiments. One-way ANOVA with Dunnet post-hoc analysis. ***p ≤ 0.001.
Figure 4 rAl-CPI stimulates Treg differentiation. (A) Percentage of Treg cells in spleen. (B) Percentage of Treg cells when the anti-IL-10R monoclonal or isotype control was used. The mean values ± SEM are shown for 6–7 mice per group; representative data from three independent experiments. One-way ANOVA with Dunnet post-hoc analysis. ***p ≤ 0.001.CC BY · PubMed Central
Figure 5 Inhibition of IL-10 signaling pathway reduces the effect of rAl-CPI on humoral response against Blomia tropicalis. (A) Total number of eosinophils and neutrophils in BAL. (B) Cytokine levels BAL, IL-10, IL-5, and IL-10. (C) Serum Total IgE, IgG1, and BtE allergen-specific IgE, IgG1, and IgG2a antibody levels. The mean values ± SEM are shown for 5–6 mice per group when the anti-IL-10R monoclonal was used. One-way ANOVA + Bonferroni for post-hoc comparisons). *p ≤ 0.05; **p ≤ 0.01, ***p ≤ 0.001.
Figure 5 Inhibition of IL-10 signaling pathway reduces the effect of rAl-CPI on humoral response against Blomia tropicalis. (A) Total number of eosinophils and neutrophils in BAL. (B) Cytokine levels BAL, IL-10, IL-5, and IL-10. (C) Serum Total IgE, IgG1, and BtE allergen-specific IgE, IgG1, and IgG2a antibody levels. The mean values ± SEM are shown for 5–6 mice per group when the anti-IL-10R monoclonal was used. One-way ANOVA + Bonferroni for post-hoc comparisons). *p ≤ 0.05; **p ≤ 0.01, ***p ≤ 0.001.CC BY · PubMed Central
Figure 6 Effects of rAl-CPI on surface molecules expression and cytokine production by HmoDCs. HmoDCs isolated from five different donors were stimulated with LPS, rAl-CPI alone o rAl-CPI+LPS and analyzed for (A) cell surface marker expression and (B) cytokine production. Data presented are means of duplicate. One-way ANOVA with Dunnet post-hoc analysis for MFI data and T-test for cytokine comparisons. *p ≤ 0.05; **p ≤ 0.01, ***p ≤ 0.001.
Figure 6 Effects of rAl-CPI on surface molecules expression and cytokine production by HmoDCs. HmoDCs isolated from five different donors were stimulated with LPS, rAl-CPI alone o rAl-CPI+LPS and analyzed for (A) cell surface marker expression and (B) cytokine production. Data presented are means of duplicate. One-way ANOVA with Dunnet post-hoc analysis for MFI data and T-test for cytokine comparisons. *p ≤ 0.05; **p ≤ 0.01, ***p ≤ 0.001.CC BY · PubMed Central

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Authors
Coronado, S., Zakzuk, J., Regino, R., Ahumada, V., Benedetti, I., Angelina, A., Palomares, O., & Caraballo, L.
Venue
Frontiers in Immunology
Publisher
Frontiers Media SA
Published
2019-09-27
Volume
10
ISSN
1664-3224
DOI
10.3389/fimmu.2019.02280
PMC
PMC6777510
Cited by
32
References
6
Licence
creativecommons.org/licenses/by/4.0/ (vor)
Funders
Departamento Administrativo de Ciencia, Tecnología e Innovación

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