IgE antibodies from schistosomiasis patients to recognize epitopes in potato apyrase

High percentages of structural identity and cross-immunoreactivity have been reported between potato apyrase and Schistosoma mansoni ATP diphosphohydrolase (SmATPDases) isoforms, showing the existence of particular epitopes shared between these proteins.

METHODS:

Potato apyrase was employed using ELISA, western blot, and mouse immunization methods to verify IgE reactivity.

RESULTS:

Most of the schistosomiasis patient’s (75%) serum was seropositive for potato apyrase and this protein was recognized using western blotting, suggesting that parasite and plant proteins share IgE-binding epitopes. C57BL/6 mice immunized with potato apyrase showed increased IgE antibody production.

CONCLUSIONS:

Potato apyrase and SmATPDases have IgE-binding epitopes.

Keywords:
ATP diphosphohydrolase; IgE; Potato apyrase

Authorship

Michélia Antônia do Nascimento Gusmão

Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brasil. Universidade Federal de Juiz de ForaBrazilJuiz de Fora, MG, BrazilInstituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brasil.

Nayara Braga Emídio

Danielle Gomes Marconato

Priscila Silva Grijó Farani

Luiz Felipe Maniezzi

Paulo Marcos Zech Coelho

Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Belo Horizonte, MG, Brasil.Fundação Oswaldo CruzBrazilBelo Horizonte, MG, BrazilCentro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Belo Horizonte, MG, Brasil.

Rodrigo Correa Oliveira

Eveline Gomes Vasconcelos

Priscila de Faria-Pinto

https://orcid.org/0000-0002-7244-6357

Corresponding author: Priscila de Faria-Pinto. e-mail:priscila.faria@ufjf.edu.br

Conflict of Interest: The authors declare that there is no conflict of interest.

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Figures

Figures (2)

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FIGURE 1:
IgE antibody reactivity of schistosomiasis patient’s serum samples to potato apyrase by ELISA. (A) IgE antibody reactivity of serum samples (diluted 1:50, tested in duplicates) of schistosomiasis patients domiciled in an endemic area (n= 102) is represented by the mean optical density (O.D.). The dotted line represents the cut-off (0.082) obtained by the mean optical density plus double the standard deviation of serum samples obtained from healthy individuals from non-endemic areas for S. mansoni (control group; n= 26). ***p < 0.001. (B) Serum samples from the schistosomiasis patients (n=20) or healthy individuals (control group; n=8) were IgG antibody-depleted. Followed by centrifugation, the supernatants were diluted 1:20 and IgE antibody reactivity with potato apyrase was tested. The dotted line represents the cut-off (0.546). **p < 0.01.

Thumbnail

FIGURE 2:
(I) IgE antibody production in C57BL/6 mice induced by potato apyrase immunization, evaluated by ELISA assay. ***p < 0.001. (II) Western blotting of potato apyrase and IgE antibody of (A) serum sample (1:50) from healthy-Swiss mice; (B) rabbit polyclonal serum anti-potato apyrase (1:3,000); (C) C57BL/6 mice polyclonal serum anti-potato apyrase (1:50); (D) serum samples (1:50) from S. mansoni-infected Swiss mice, and (E) pool of serum samples from patients with schistosomiasis. Signals were revealed by chemiluminescence using isotype-specific anti-rabbit IgG (B) or anti-mouse IgE (A, C, D, E) coupled to horseradish peroxidase, luminol as substrate (ECL Western Blotting System; GE Healthcare, Brazil), and can be detected by X-ray film.

FIGURE 1: IgE antibody reactivity of schistosomiasis patient’s serum samples to potato apyrase by ELISA. (A) IgE antibody reactivity of serum samples (diluted 1:50, tested in duplicates) of schistosomiasis patients domiciled in an endemic area (n= 102) is represented by the mean optical density (O.D.). The dotted line represents the cut-off (0.082) obtained by the mean optical density plus double the standard deviation of serum samples obtained from healthy individuals from non-endemic areas for S. mansoni (control group; n= 26). *p < 0.001. (B) Serum samples from the schistosomiasis patients (n=20) or healthy individuals (control group; n=8) were IgG antibody-depleted. Followed by centrifugation, the supernatants were diluted 1:20 and IgE antibody reactivity with potato apyrase was tested. The dotted line represents the cut-off (0.546). p < 0.01.

FIGURE 2: (I) IgE antibody production in C57BL/6 mice induced by potato apyrase immunization, evaluated by ELISA assay. ***p < 0.001. (II) Western blotting of potato apyrase and IgE antibody of (A) serum sample (1:50) from healthy-Swiss mice; (B) rabbit polyclonal serum anti-potato apyrase (1:3,000); (C) C57BL/6 mice polyclonal serum anti-potato apyrase (1:50); (D) serum samples (1:50) from S. mansoni-infected Swiss mice, and (E) pool of serum samples from patients with schistosomiasis. Signals were revealed by chemiluminescence using isotype-specific anti-rabbit IgG (B) or anti-mouse IgE (A, C, D, E) coupled to horseradish peroxidase, luminol as substrate (ECL Western Blotting System; GE Healthcare, Brazil), and can be detected by X-ray film.

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IgE antibodies from schistosomiasis patients to recognize epitopes in potato apyrase

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[1] Corresponding author: Priscila de Faria-Pinto. e-mail:priscila.faria@ufjf.edu.br