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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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Immunological and biochemical characterization of ESP from S. mansoni eggs. (A) Monocyte-derived DCs pulsed for 48 h with the different antigen preparations in the absence (top) or presence (bottom) of 100 ng/ml LPS as a maturation factor were co-cultured with allogeneic naive CD4+ T cells for 12 d in the presence of staphylococcal enterotoxin B and IL-2. Intracellular cytokine production was assayed by FACS 6 h after the stimulation of primed T cells with PMA and ionomycin. The frequencies of each population are indicated as percentages in the plot. One representative result from three independent experiments is shown. (B) 5 µg/cm ESP was separated under nonreducing conditions by SDS-PAGE and silver stained. (C) The presence of omega-1 and IPSE/alpha-1 was confirmed on Western blots by staining with specific anti–IPSE/alpha-1 and anti–omega-1 monoclonal antibodies.
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fig1: Immunological and biochemical characterization of ESP from S. mansoni eggs. (A) Monocyte-derived DCs pulsed for 48 h with the different antigen preparations in the absence (top) or presence (bottom) of 100 ng/ml LPS as a maturation factor were co-cultured with allogeneic naive CD4+ T cells for 12 d in the presence of staphylococcal enterotoxin B and IL-2. Intracellular cytokine production was assayed by FACS 6 h after the stimulation of primed T cells with PMA and ionomycin. The frequencies of each population are indicated as percentages in the plot. One representative result from three independent experiments is shown. (B) 5 µg/cm ESP was separated under nonreducing conditions by SDS-PAGE and silver stained. (C) The presence of omega-1 and IPSE/alpha-1 was confirmed on Western blots by staining with specific anti–IPSE/alpha-1 and anti–omega-1 monoclonal antibodies.

Mentions: DCs are known to play a pivotal role in the initiation and polarization of T cell responses, and S. mansoni egg preparations have been shown to prime Th2 cells via the functional modulation of DCs (MacDonald et al., 2001; de Jong et al., 2002). To study and identify the components from S. mansoni egg preparations that instruct Th2 development, we used a well established co-culture system of human monocyte-derived DC and naive CD4+ T cells, which is generally thought to mimic in vivo DC-mediated T helper cell polarization (Kapsenberg, 2003). It stands to reason that ESPs from live eggs (Cass et al., 2007) are the first egg-derived molecules to interact with cells of the innate immune system, including DCs. Therefore, we initially tested ESPs for their capacity to condition DCs to prime Th2 development from naive CD4+ T cells. Similar to SEA, exposure of DCs to ESP resulted in a robust Th2 skewing irrespective of the presence or absence of LPS as a neutral maturation factor (Fig. 1 A). In a recent paper, Cass et al. (2007) identified omega-1 and IPSE/alpha-1 as the most abundant proteins within ESP from S. mansoni eggs. Separation of ESP preparations by SDS-PAGE (Fig. 1 B), followed by Western blotting with specific monoclonal antibodies, revealed prominent bands representing IPSE/alpha-1 and omega-1 (Fig. 1 C), which was confirmed by mass spectrometry (not depicted). Both omega-1 and IPSE/alpha-1 are glycoproteins that are specifically expressed in and secreted from S. mansoni eggs. Omega-1 has been demonstrated to display RNase activity and hepatotoxic effects (Dunne et al., 1991; Fitzsimmons et al., 2005), whereas IPSE/alpha-1 has previously been shown to trigger IL-4 production by human and mouse basophils (Schramm et al., 2003, 2007).


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)

Immunological and biochemical characterization of ESP from S. mansoni eggs. (A) Monocyte-derived DCs pulsed for 48 h with the different antigen preparations in the absence (top) or presence (bottom) of 100 ng/ml LPS as a maturation factor were co-cultured with allogeneic naive CD4+ T cells for 12 d in the presence of staphylococcal enterotoxin B and IL-2. Intracellular cytokine production was assayed by FACS 6 h after the stimulation of primed T cells with PMA and ionomycin. The frequencies of each population are indicated as percentages in the plot. One representative result from three independent experiments is shown. (B) 5 µg/cm ESP was separated under nonreducing conditions by SDS-PAGE and silver stained. (C) The presence of omega-1 and IPSE/alpha-1 was confirmed on Western blots by staining with specific anti–IPSE/alpha-1 and anti–omega-1 monoclonal antibodies.
© Copyright Policy - openaccess
Related In: Results  -  Collection

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

fig1: Immunological and biochemical characterization of ESP from S. mansoni eggs. (A) Monocyte-derived DCs pulsed for 48 h with the different antigen preparations in the absence (top) or presence (bottom) of 100 ng/ml LPS as a maturation factor were co-cultured with allogeneic naive CD4+ T cells for 12 d in the presence of staphylococcal enterotoxin B and IL-2. Intracellular cytokine production was assayed by FACS 6 h after the stimulation of primed T cells with PMA and ionomycin. The frequencies of each population are indicated as percentages in the plot. One representative result from three independent experiments is shown. (B) 5 µg/cm ESP was separated under nonreducing conditions by SDS-PAGE and silver stained. (C) The presence of omega-1 and IPSE/alpha-1 was confirmed on Western blots by staining with specific anti–IPSE/alpha-1 and anti–omega-1 monoclonal antibodies.
Mentions: DCs are known to play a pivotal role in the initiation and polarization of T cell responses, and S. mansoni egg preparations have been shown to prime Th2 cells via the functional modulation of DCs (MacDonald et al., 2001; de Jong et al., 2002). To study and identify the components from S. mansoni egg preparations that instruct Th2 development, we used a well established co-culture system of human monocyte-derived DC and naive CD4+ T cells, which is generally thought to mimic in vivo DC-mediated T helper cell polarization (Kapsenberg, 2003). It stands to reason that ESPs from live eggs (Cass et al., 2007) are the first egg-derived molecules to interact with cells of the innate immune system, including DCs. Therefore, we initially tested ESPs for their capacity to condition DCs to prime Th2 development from naive CD4+ T cells. Similar to SEA, exposure of DCs to ESP resulted in a robust Th2 skewing irrespective of the presence or absence of LPS as a neutral maturation factor (Fig. 1 A). In a recent paper, Cass et al. (2007) identified omega-1 and IPSE/alpha-1 as the most abundant proteins within ESP from S. mansoni eggs. Separation of ESP preparations by SDS-PAGE (Fig. 1 B), followed by Western blotting with specific monoclonal antibodies, revealed prominent bands representing IPSE/alpha-1 and omega-1 (Fig. 1 C), which was confirmed by mass spectrometry (not depicted). Both omega-1 and IPSE/alpha-1 are glycoproteins that are specifically expressed in and secreted from S. mansoni eggs. Omega-1 has been demonstrated to display RNase activity and hepatotoxic effects (Dunne et al., 1991; Fitzsimmons et al., 2005), whereas IPSE/alpha-1 has previously been shown to trigger IL-4 production by human and mouse basophils (Schramm et al., 2003, 2007).

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