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Glycans from Fasciola hepatica Modulate the Host Immune Response and TLR-Induced Maturation of Dendritic Cells.

Rodríguez E, Noya V, Cervi L, Chiribao ML, Brossard N, Chiale C, Carmona C, Giacomini C, Freire T - PLoS Negl Trop Dis (2015)

Bottom Line: Our results indicate that glycans from F. hepatica promote the production of IL-4 and IL-10, suppressing IFNγ production.Inhibition assays using carbohydrates suggest that the immune-modulation is mediated, at least in part, by the recognition of a mannose specific-CLR that signals by recruiting the phosphatase Php2.The results presented here contribute to the understanding of the role of parasite glycosylated molecules in the modulation of the host immunity and might be useful in the design of vaccines against fasciolosis.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunomodulation and Vaccine Development, Departamento de Inmunobiología, Facultad de Medicina, UdelaR, Montevideo, Uruguay.

ABSTRACT
Helminths express various carbohydrate-containing glycoconjugates on their surface, and they release glycan-rich excretion/secretion products that can be very important in their life cycles, infection and pathology. Recent evidence suggests that parasite glycoconjugates could play a role in the evasion of the immune response, leading to a modified Th2-polarized immune response that favors parasite survival in the host. Nevertheless, there is limited information about the nature or function of glycans produced by the trematode Fasciola hepatica, the causative agent of fasciolosis. In this paper, we investigate whether glycosylated molecules from F. hepatica participate in the modulation of host immunity. We also focus on dendritic cells, since they are an important target of immune-modulation by helminths, affecting their activity or function. Our results indicate that glycans from F. hepatica promote the production of IL-4 and IL-10, suppressing IFNγ production. During infection, this parasite is able to induce a semi-mature phenotype of DCs expressing low levels of MHCII and secrete IL-10. Furthermore, we show that parasite glycoconjugates mediate the modulation of LPS-induced maturation of DCs since their oxidation restores the capacity of LPS-treated DCs to secrete high levels of the pro-inflammatory cytokines IL-6 and IL-12/23p40 and low levels of the anti-inflammatory cytokine IL-10. Inhibition assays using carbohydrates suggest that the immune-modulation is mediated, at least in part, by the recognition of a mannose specific-CLR that signals by recruiting the phosphatase Php2. The results presented here contribute to the understanding of the role of parasite glycosylated molecules in the modulation of the host immunity and might be useful in the design of vaccines against fasciolosis.

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Oxidation of parasite components partially inhibits modulation of LPS-induced maturation of DCs by FhTE.BMDCs were cultured in the presence of 75 μg/mL of FhTE, FhCB (oxidation negative control) or FhmPox (oxidized FhTE) in presence or absence of LPS (1 μg/ml) overnight at 37°C. Then, culture supernatants were collected and analyzed by ELISA for IL-6, IL-10 or IL-12/23p40 (A). BMDCs were also pre-incubated for 45 min. with mannose (Man), N-Acetyl-Galactosamine (GalNAc) or arabinose (Ara) at 10 mM and then stimulated as in A. Culture supernatants were analyzed by ELISA for detection of IL-6, IL-10 or IL-12/23p40 (B) and MIP-1α and MIP-2 (C). Alternatively, BMDCs were pre-incubated for 45 min. with 10 μM of specific signaling inhibitors (PHPS1; GW5074; and ER27319) and then stimulated with FhTE (75 μg/mL) in presence of LPS (1 μg/ml) (D). Results are expressed as the mean of three independent experiments (±SD, indicated by error bars). Asterisks indicate statistically significant differences (*p < 0.01) with respect to LPS-stimulated BMDCs.
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pntd.0004234.g007: Oxidation of parasite components partially inhibits modulation of LPS-induced maturation of DCs by FhTE.BMDCs were cultured in the presence of 75 μg/mL of FhTE, FhCB (oxidation negative control) or FhmPox (oxidized FhTE) in presence or absence of LPS (1 μg/ml) overnight at 37°C. Then, culture supernatants were collected and analyzed by ELISA for IL-6, IL-10 or IL-12/23p40 (A). BMDCs were also pre-incubated for 45 min. with mannose (Man), N-Acetyl-Galactosamine (GalNAc) or arabinose (Ara) at 10 mM and then stimulated as in A. Culture supernatants were analyzed by ELISA for detection of IL-6, IL-10 or IL-12/23p40 (B) and MIP-1α and MIP-2 (C). Alternatively, BMDCs were pre-incubated for 45 min. with 10 μM of specific signaling inhibitors (PHPS1; GW5074; and ER27319) and then stimulated with FhTE (75 μg/mL) in presence of LPS (1 μg/ml) (D). Results are expressed as the mean of three independent experiments (±SD, indicated by error bars). Asterisks indicate statistically significant differences (*p < 0.01) with respect to LPS-stimulated BMDCs.

Mentions: In order to evaluate whether parasite glycosylated molecules are able to influence DC-maturation we incubated BMDCs with parasite components or oxidized-FhTE in absence or presence of a maturation stimulus (LPS). Furthermore, the control FhCB (consisting in FhTE subjected to the whole treatment excepting for the incubation with sodium periodate) was also included. Then, we evaluated the production of several cytokines by DCs. BMDCs incubated with FhTE in presence of LPS produced higher levels of IL-10 than DCs incubated only with LPS. Interestingly, when oxidized-parasite glycans (FhmPox) where incubated with BMDCs, they produced similar levels of IL-10 than those produced by cells incubated with LPS alone, indicating that oxidation of glycans abrogates the immunomodulatory activity of FhTE on DCs (Fig 7A). On the other hand, the opposite situation was observed with the pro-inflammatory cytokines IL-6 and IL-12/23p40. Indeed, FhTE incubated with BMDCs in presence of LPS decreased the production of IL-6 and IL-12/23p40 induced by LPS, while FhTE oxidation restored the levels of both cytokines (Fig 7A). As expected, the control FhCB/LPS behaved essentially in the same way as FhTE/LPS, while the CmPox/LPS condition induced the same levels of cytokine production than cells incubated only with LPS. Altogether, these results indicate that glycoconjugates from F. hepatica modulate LPS-induced maturation of DCs by augmenting anti-inflammatory cytokines and decreasing pro-inflammatory cytokines.


Glycans from Fasciola hepatica Modulate the Host Immune Response and TLR-Induced Maturation of Dendritic Cells.

Rodríguez E, Noya V, Cervi L, Chiribao ML, Brossard N, Chiale C, Carmona C, Giacomini C, Freire T - PLoS Negl Trop Dis (2015)

Oxidation of parasite components partially inhibits modulation of LPS-induced maturation of DCs by FhTE.BMDCs were cultured in the presence of 75 μg/mL of FhTE, FhCB (oxidation negative control) or FhmPox (oxidized FhTE) in presence or absence of LPS (1 μg/ml) overnight at 37°C. Then, culture supernatants were collected and analyzed by ELISA for IL-6, IL-10 or IL-12/23p40 (A). BMDCs were also pre-incubated for 45 min. with mannose (Man), N-Acetyl-Galactosamine (GalNAc) or arabinose (Ara) at 10 mM and then stimulated as in A. Culture supernatants were analyzed by ELISA for detection of IL-6, IL-10 or IL-12/23p40 (B) and MIP-1α and MIP-2 (C). Alternatively, BMDCs were pre-incubated for 45 min. with 10 μM of specific signaling inhibitors (PHPS1; GW5074; and ER27319) and then stimulated with FhTE (75 μg/mL) in presence of LPS (1 μg/ml) (D). Results are expressed as the mean of three independent experiments (±SD, indicated by error bars). Asterisks indicate statistically significant differences (*p < 0.01) with respect to LPS-stimulated BMDCs.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4697847&req=5

pntd.0004234.g007: Oxidation of parasite components partially inhibits modulation of LPS-induced maturation of DCs by FhTE.BMDCs were cultured in the presence of 75 μg/mL of FhTE, FhCB (oxidation negative control) or FhmPox (oxidized FhTE) in presence or absence of LPS (1 μg/ml) overnight at 37°C. Then, culture supernatants were collected and analyzed by ELISA for IL-6, IL-10 or IL-12/23p40 (A). BMDCs were also pre-incubated for 45 min. with mannose (Man), N-Acetyl-Galactosamine (GalNAc) or arabinose (Ara) at 10 mM and then stimulated as in A. Culture supernatants were analyzed by ELISA for detection of IL-6, IL-10 or IL-12/23p40 (B) and MIP-1α and MIP-2 (C). Alternatively, BMDCs were pre-incubated for 45 min. with 10 μM of specific signaling inhibitors (PHPS1; GW5074; and ER27319) and then stimulated with FhTE (75 μg/mL) in presence of LPS (1 μg/ml) (D). Results are expressed as the mean of three independent experiments (±SD, indicated by error bars). Asterisks indicate statistically significant differences (*p < 0.01) with respect to LPS-stimulated BMDCs.
Mentions: In order to evaluate whether parasite glycosylated molecules are able to influence DC-maturation we incubated BMDCs with parasite components or oxidized-FhTE in absence or presence of a maturation stimulus (LPS). Furthermore, the control FhCB (consisting in FhTE subjected to the whole treatment excepting for the incubation with sodium periodate) was also included. Then, we evaluated the production of several cytokines by DCs. BMDCs incubated with FhTE in presence of LPS produced higher levels of IL-10 than DCs incubated only with LPS. Interestingly, when oxidized-parasite glycans (FhmPox) where incubated with BMDCs, they produced similar levels of IL-10 than those produced by cells incubated with LPS alone, indicating that oxidation of glycans abrogates the immunomodulatory activity of FhTE on DCs (Fig 7A). On the other hand, the opposite situation was observed with the pro-inflammatory cytokines IL-6 and IL-12/23p40. Indeed, FhTE incubated with BMDCs in presence of LPS decreased the production of IL-6 and IL-12/23p40 induced by LPS, while FhTE oxidation restored the levels of both cytokines (Fig 7A). As expected, the control FhCB/LPS behaved essentially in the same way as FhTE/LPS, while the CmPox/LPS condition induced the same levels of cytokine production than cells incubated only with LPS. Altogether, these results indicate that glycoconjugates from F. hepatica modulate LPS-induced maturation of DCs by augmenting anti-inflammatory cytokines and decreasing pro-inflammatory cytokines.

Bottom Line: Our results indicate that glycans from F. hepatica promote the production of IL-4 and IL-10, suppressing IFNγ production.Inhibition assays using carbohydrates suggest that the immune-modulation is mediated, at least in part, by the recognition of a mannose specific-CLR that signals by recruiting the phosphatase Php2.The results presented here contribute to the understanding of the role of parasite glycosylated molecules in the modulation of the host immunity and might be useful in the design of vaccines against fasciolosis.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunomodulation and Vaccine Development, Departamento de Inmunobiología, Facultad de Medicina, UdelaR, Montevideo, Uruguay.

ABSTRACT
Helminths express various carbohydrate-containing glycoconjugates on their surface, and they release glycan-rich excretion/secretion products that can be very important in their life cycles, infection and pathology. Recent evidence suggests that parasite glycoconjugates could play a role in the evasion of the immune response, leading to a modified Th2-polarized immune response that favors parasite survival in the host. Nevertheless, there is limited information about the nature or function of glycans produced by the trematode Fasciola hepatica, the causative agent of fasciolosis. In this paper, we investigate whether glycosylated molecules from F. hepatica participate in the modulation of host immunity. We also focus on dendritic cells, since they are an important target of immune-modulation by helminths, affecting their activity or function. Our results indicate that glycans from F. hepatica promote the production of IL-4 and IL-10, suppressing IFNγ production. During infection, this parasite is able to induce a semi-mature phenotype of DCs expressing low levels of MHCII and secrete IL-10. Furthermore, we show that parasite glycoconjugates mediate the modulation of LPS-induced maturation of DCs since their oxidation restores the capacity of LPS-treated DCs to secrete high levels of the pro-inflammatory cytokines IL-6 and IL-12/23p40 and low levels of the anti-inflammatory cytokine IL-10. Inhibition assays using carbohydrates suggest that the immune-modulation is mediated, at least in part, by the recognition of a mannose specific-CLR that signals by recruiting the phosphatase Php2. The results presented here contribute to the understanding of the role of parasite glycosylated molecules in the modulation of the host immunity and might be useful in the design of vaccines against fasciolosis.

Show MeSH
Related in: MedlinePlus