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Omega-1, a glycoprotein secreted by Schistosoma mansoni eggs, drives Th2 responses.

Everts B, Perona-Wright G, Smits HH, Hokke CH, van der Ham AJ, Fitzsimmons CM, Doenhoff MJ, van der Bosch J, Mohrs K, Haas H, Mohrs M, Yazdanbakhsh M, Schramm G - J. Exp. Med. (2009)

Bottom Line: We report that omega-1, a glycoprotein which is secreted from S. mansoni eggs and present in SEA, is capable of conditioning human monocyte-derived dendritic cells in vitro to drive T helper 2 (Th2) polarization with similar characteristics as whole SEA.Finally, omega-1-depleted SEA displays an impaired capacity for Th2 priming in vitro, but not in vivo, suggesting the existence of additional factors within SEA that can compensate for the omega-1-mediated effects.Collectively, we identify omega-1, a single component of SEA, as a potent inducer of Th2 responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Parasitology, Leiden University Medical Centre, Leiden 2333 ZA, The Netherlands.

ABSTRACT
Soluble egg antigens of the parasitic helminth Schistosoma mansoni (S. mansoni egg antigen [SEA]) induce strong Th2 responses both in vitro and in vivo. However, the specific molecules that prime the development of Th2 responses have not been identified. We report that omega-1, a glycoprotein which is secreted from S. mansoni eggs and present in SEA, is capable of conditioning human monocyte-derived dendritic cells in vitro to drive T helper 2 (Th2) polarization with similar characteristics as whole SEA. Furthermore, using IL-4 dual reporter mice, we show that both natural and recombinant omega-1 alone are sufficient to generate Th2 responses in vivo, even in the absence of IL-4R signaling. Finally, omega-1-depleted SEA displays an impaired capacity for Th2 priming in vitro, but not in vivo, suggesting the existence of additional factors within SEA that can compensate for the omega-1-mediated effects. Collectively, we identify omega-1, a single component of SEA, as a potent inducer of Th2 responses.

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SDS-PAGE of SEA, natural omega-1, and natural IPSE/alpha-1 as well as of recombinant omega-1 (silver staining and Western blotting). (A–C) 5 µg/cm SEA, 0.3 µg/cm omega-1, 0.3 µg/cm and IPSE/alpha-1 purified from SEA were separated by SDS-PAGE and silver stained or blotted onto nitrocellulose membrane. Silver staining (A) revealed a weak banding intensity of both natural and recombinant omega-1 compared with IPSE/alpha-1, although the purified proteins were applied to the gel at the same amounts (0.3 µg/cm). The two bands stained by anti-IPSE/alpha-1 represent posttranslational variants of the same protein (Schramm et al., 2003). On Western blots, alkaline phosphatase–labeled A. aurantia agglutinin (B) or a mixture of specific anti–IPSE/alpha-1 and anti–omega-1 monoclonal antibodies followed by alkaline phosphatase–labeled anti–mouse IgG secondary antibody (C) were used for detection. (B) Although A. aurantia agglutinin clearly binds to omega-1 and IPSE/alpha-1 as well as to a variety of other fucosylated components present in SEA, it does not bind to recombinant omega-1, whose glycans are lacking fucose residues (note that the double band of recombinant vs. natural omega-1 is caused by differential glycosylation). (C) In contrast, all purified proteins but no irrelevant SEA components are detected by the mixture of specific monoclonal antibodies.
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fig2: SDS-PAGE of SEA, natural omega-1, and natural IPSE/alpha-1 as well as of recombinant omega-1 (silver staining and Western blotting). (A–C) 5 µg/cm SEA, 0.3 µg/cm omega-1, 0.3 µg/cm and IPSE/alpha-1 purified from SEA were separated by SDS-PAGE and silver stained or blotted onto nitrocellulose membrane. Silver staining (A) revealed a weak banding intensity of both natural and recombinant omega-1 compared with IPSE/alpha-1, although the purified proteins were applied to the gel at the same amounts (0.3 µg/cm). The two bands stained by anti-IPSE/alpha-1 represent posttranslational variants of the same protein (Schramm et al., 2003). On Western blots, alkaline phosphatase–labeled A. aurantia agglutinin (B) or a mixture of specific anti–IPSE/alpha-1 and anti–omega-1 monoclonal antibodies followed by alkaline phosphatase–labeled anti–mouse IgG secondary antibody (C) were used for detection. (B) Although A. aurantia agglutinin clearly binds to omega-1 and IPSE/alpha-1 as well as to a variety of other fucosylated components present in SEA, it does not bind to recombinant omega-1, whose glycans are lacking fucose residues (note that the double band of recombinant vs. natural omega-1 is caused by differential glycosylation). (C) In contrast, all purified proteins but no irrelevant SEA components are detected by the mixture of specific monoclonal antibodies.

Mentions: The observation that ESP can instruct human DCs to drive highly polarized Th2 responses prompted the question of whether omega-1 and IPSE/alpha-1 as prominent ESP components are responsible for this activity. Although some immunological properties of IPSE/alpha-1 have been described (Schramm et al., 2003, 2007), the effects of omega-1 and IPSE/alpha-1 on DC-driven T helper cell polarization have not been investigated. To this end, natural omega-1 and IPSE/alpha-1 were purified from SEA (Fig. 2) and used for the conditioning of human DCs in comparison with whole SEA. The concentrations of omega-1 and IPSE/alpha-1 used in these assays were equivalent to those in the unfractionated SEA preparations. As described previously (Kane et al., 2004; van Liempt et al., 2007), stimulation with SEA did not lead to classical maturation of DCs, based on surface marker expression (Fig. S1). Likewise, omega-1 and IPSE/alpha-1 did not induce the expression of these markers on DCs (Fig. S1). Apart from the failure to induce the maturation of DCs, SEA is also known to interfere with TLR-mediated DC activation (Kane et al., 2004; van Liempt et al., 2007). Indeed, when DCs were matured with the TLR4 ligand LPS, a nonpolarizing maturation factor for human DCs, the presence of SEA significantly impaired the LPS-induced up-regulation of CD83 and CD86 surface expression (Fig. 3 A). Strikingly, omega-1 alone was sufficient to suppress the induction of these molecules on LPS-stimulated DCs to a similar extent, whereas IPSE/alpha-1 had no effect (Fig. 3 A).


Omega-1, a glycoprotein secreted by Schistosoma mansoni eggs, drives Th2 responses.

Everts B, Perona-Wright G, Smits HH, Hokke CH, van der Ham AJ, Fitzsimmons CM, Doenhoff MJ, van der Bosch J, Mohrs K, Haas H, Mohrs M, Yazdanbakhsh M, Schramm G - J. Exp. Med. (2009)

SDS-PAGE of SEA, natural omega-1, and natural IPSE/alpha-1 as well as of recombinant omega-1 (silver staining and Western blotting). (A–C) 5 µg/cm SEA, 0.3 µg/cm omega-1, 0.3 µg/cm and IPSE/alpha-1 purified from SEA were separated by SDS-PAGE and silver stained or blotted onto nitrocellulose membrane. Silver staining (A) revealed a weak banding intensity of both natural and recombinant omega-1 compared with IPSE/alpha-1, although the purified proteins were applied to the gel at the same amounts (0.3 µg/cm). The two bands stained by anti-IPSE/alpha-1 represent posttranslational variants of the same protein (Schramm et al., 2003). On Western blots, alkaline phosphatase–labeled A. aurantia agglutinin (B) or a mixture of specific anti–IPSE/alpha-1 and anti–omega-1 monoclonal antibodies followed by alkaline phosphatase–labeled anti–mouse IgG secondary antibody (C) were used for detection. (B) Although A. aurantia agglutinin clearly binds to omega-1 and IPSE/alpha-1 as well as to a variety of other fucosylated components present in SEA, it does not bind to recombinant omega-1, whose glycans are lacking fucose residues (note that the double band of recombinant vs. natural omega-1 is caused by differential glycosylation). (C) In contrast, all purified proteins but no irrelevant SEA components are detected by the mixture of specific monoclonal antibodies.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2722183&req=5

fig2: SDS-PAGE of SEA, natural omega-1, and natural IPSE/alpha-1 as well as of recombinant omega-1 (silver staining and Western blotting). (A–C) 5 µg/cm SEA, 0.3 µg/cm omega-1, 0.3 µg/cm and IPSE/alpha-1 purified from SEA were separated by SDS-PAGE and silver stained or blotted onto nitrocellulose membrane. Silver staining (A) revealed a weak banding intensity of both natural and recombinant omega-1 compared with IPSE/alpha-1, although the purified proteins were applied to the gel at the same amounts (0.3 µg/cm). The two bands stained by anti-IPSE/alpha-1 represent posttranslational variants of the same protein (Schramm et al., 2003). On Western blots, alkaline phosphatase–labeled A. aurantia agglutinin (B) or a mixture of specific anti–IPSE/alpha-1 and anti–omega-1 monoclonal antibodies followed by alkaline phosphatase–labeled anti–mouse IgG secondary antibody (C) were used for detection. (B) Although A. aurantia agglutinin clearly binds to omega-1 and IPSE/alpha-1 as well as to a variety of other fucosylated components present in SEA, it does not bind to recombinant omega-1, whose glycans are lacking fucose residues (note that the double band of recombinant vs. natural omega-1 is caused by differential glycosylation). (C) In contrast, all purified proteins but no irrelevant SEA components are detected by the mixture of specific monoclonal antibodies.
Mentions: The observation that ESP can instruct human DCs to drive highly polarized Th2 responses prompted the question of whether omega-1 and IPSE/alpha-1 as prominent ESP components are responsible for this activity. Although some immunological properties of IPSE/alpha-1 have been described (Schramm et al., 2003, 2007), the effects of omega-1 and IPSE/alpha-1 on DC-driven T helper cell polarization have not been investigated. To this end, natural omega-1 and IPSE/alpha-1 were purified from SEA (Fig. 2) and used for the conditioning of human DCs in comparison with whole SEA. The concentrations of omega-1 and IPSE/alpha-1 used in these assays were equivalent to those in the unfractionated SEA preparations. As described previously (Kane et al., 2004; van Liempt et al., 2007), stimulation with SEA did not lead to classical maturation of DCs, based on surface marker expression (Fig. S1). Likewise, omega-1 and IPSE/alpha-1 did not induce the expression of these markers on DCs (Fig. S1). Apart from the failure to induce the maturation of DCs, SEA is also known to interfere with TLR-mediated DC activation (Kane et al., 2004; van Liempt et al., 2007). Indeed, when DCs were matured with the TLR4 ligand LPS, a nonpolarizing maturation factor for human DCs, the presence of SEA significantly impaired the LPS-induced up-regulation of CD83 and CD86 surface expression (Fig. 3 A). Strikingly, omega-1 alone was sufficient to suppress the induction of these molecules on LPS-stimulated DCs to a similar extent, whereas IPSE/alpha-1 had no effect (Fig. 3 A).

Bottom Line: We report that omega-1, a glycoprotein which is secreted from S. mansoni eggs and present in SEA, is capable of conditioning human monocyte-derived dendritic cells in vitro to drive T helper 2 (Th2) polarization with similar characteristics as whole SEA.Finally, omega-1-depleted SEA displays an impaired capacity for Th2 priming in vitro, but not in vivo, suggesting the existence of additional factors within SEA that can compensate for the omega-1-mediated effects.Collectively, we identify omega-1, a single component of SEA, as a potent inducer of Th2 responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Parasitology, Leiden University Medical Centre, Leiden 2333 ZA, The Netherlands.

ABSTRACT
Soluble egg antigens of the parasitic helminth Schistosoma mansoni (S. mansoni egg antigen [SEA]) induce strong Th2 responses both in vitro and in vivo. However, the specific molecules that prime the development of Th2 responses have not been identified. We report that omega-1, a glycoprotein which is secreted from S. mansoni eggs and present in SEA, is capable of conditioning human monocyte-derived dendritic cells in vitro to drive T helper 2 (Th2) polarization with similar characteristics as whole SEA. Furthermore, using IL-4 dual reporter mice, we show that both natural and recombinant omega-1 alone are sufficient to generate Th2 responses in vivo, even in the absence of IL-4R signaling. Finally, omega-1-depleted SEA displays an impaired capacity for Th2 priming in vitro, but not in vivo, suggesting the existence of additional factors within SEA that can compensate for the omega-1-mediated effects. Collectively, we identify omega-1, a single component of SEA, as a potent inducer of Th2 responses.

Show MeSH
Related in: MedlinePlus