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Anti-Ascaris suum immunoglobulin Y as a novel biotechnological tool for the diagnosis of human ascariasis

Published online by Cambridge University Press:  14 August 2019

C.A. Lopes ,
L.S. de Faria ,
J.E.N. de Sousa ,
I.P. Borges ,
R.P. Ribeiro ,
L.L. Bueno ,
V.M. Rodrigues Ávila ,
Á. Ferreira Júnior  and
J.M. Costa-Cruz Open the ORCID record for J.M. Costa-Cruz [Opens in a new window]
C.A. Lopes
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
L.S. de Faria
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
J.E.N. de Sousa
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
I.P. Borges
Affiliation:
Laboratório de Bioquímica e Toxinas Animais, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
R.P. Ribeiro
Affiliation:
Programa de Pós-Graduação em Sanidade e Produção Animal nos Trópicos, Universidade de Uberaba, Campus Aeroporto. Av. Nenê Sabino, 1802, sala 2D05, Uberaba, Minas Gerais 38055-500, Brazil
L.L. Bueno
Affiliation:
Laboratório de Imunologia e Genômica de Parasitos, Departamento de Parasitologia, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, Belo Horizonte, Minas Gerais 31270-901, Brazil
V.M. Rodrigues Ávila
Affiliation:
Laboratório de Bioquímica e Toxinas Animais, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
Á. Ferreira Júnior
Affiliation:
Programa de Pós-Graduação em Sanidade e Produção Animal nos Trópicos, Universidade de Uberaba, Campus Aeroporto. Av. Nenê Sabino, 1802, sala 2D05, Uberaba, Minas Gerais 38055-500, Brazil
J.M. Costa-Cruz*
Affiliation:
Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Av. Pará, 1720, Uberlândia, Minas Gerais 38400-902, Brazil
*
Author for correspondence: J.M. Costa-Cruz, E-mail: [email protected]
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Abstract

Human ascariasis is a neglected tropical disease of great relevance to public health and is considered the most frequent helminthiasis in poor regions. Accurately diagnosing this parasite has been challenging due to limitations of current diagnostic methods. Immunoglobulin Y (IgY) technology is a very effective alternative for the production of highly specific and profitable antibodies. This study aimed to produce and apply anti-Ascaris suum IgY antibodies in the immunodiagnosis of human ascariasis. Five immunizations comprising total saline extract from A. suum adult life forms were given at 14-day intervals to Gallus gallus domesticus hens of the Isa Brown line. Eggs and blood samples were collected weekly and fortnightly, respectively, to monitor the production of antibodies. The specificity of antibodies was confirmed by dot-blot, kinetic enzyme-linked immunosorbent assay (ELISA), avidity ELISA, immunoblotting and indirect immunofluorescence antibody tests. The application for disease diagnosis was performed through the detection of immune complexes in human serum samples by sandwich ELISA. Peaks of IgY anti-A. suum production occurred at weeks 6 and 8. IgY showed high avidity levels after the second dose of immunization, ranging from 64% to 93%, with a mean avidity index of 78.30%. Purified IgY recognized 12 bands of proteins from A. suum saline extract. Eggs, the uterine portion and cuticles of A. suum female adult are reactive in immunofluorescence. The detection of immune complexes showed diagnostic values of 80% sensitivity and 90% specificity. In conclusion, specific IgY have been shown to be a potential immunodiagnostic tool with promising future applications in human ascariasis.

Keywords

Human ascariasisIgYimmune compleximmunoblottingimmunodiagnosisimmunofluorescence
Type
Research Paper
Information
Journal of Helminthology , Volume 94 , 2020 , e71
Copyright
Copyright © Cambridge University Press 2019 

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References

Akita, EM and Nakai, S (1993) Comparation of four purification methods for the production of immunoglogulins from eggs laid by hens immunized with a enterotoxigenic E. coli strain. Journal of Immunological Methods 160, 207–214.Google Scholar
Anderson, TJ (1995) Ascaris infections in humans from North America: molecular evidence for cross-infection. Parasitology 110, 215–219.Google Scholar
Andrade, AM, Perez, Y, Lopez, C, Collazos, SS, Andrade, AM, Ramirez, GO and Andrade, LM (2015) Intestinal obstruction in a 3-year-old girl by Ascaris lumbricoides infestation: case report and review of the literature. Medicine 94, e655.Google Scholar
Baermann, G (1917) Eine Einfache Methode zur Auffindung von Ankylostomum (Nematoden) Larven in Erdproben. Mededeel. Mith H. Geneesk. Batavia. Lab Weltevreden Feestbundel, 41–47.Google Scholar
Bharti, B, Bharti, S and Khurana, S (2018) Worm infestation: diagnosis, treatment and prevention. Indian Journal of Pediatrics 85, 1017–1024.Google Scholar
Borges, IP, Silva, MF, Santiago, FM, et al. (2018) Antiparasitic effects induced by polyclonal IgY antibodies anti-phospholipase A2 from Bothrops pauloensis venom. International Journal of Biological Macromolecules 112, 333–342.Google Scholar
Carlander, D, Kollberg, H, Wejáker, PE and Larsson, A (2000) Peroral immunotherapy with yolk antibodies for the prevention and treatment of enteric infections. Immunologic Research 21, 1–6.Google Scholar
Claus, PE, Ceuppens, AS, Cool, M and Alliet, G (2018) Ascaris lumbricoides: challenges in diagnosis, treatment and prevention strategies in a European refugee camp. Acta Clinica Belgica 73, 431–434.Google Scholar
Da Silva Alves, EB, Conceição, MJ and Leles, D (2016) Ascaris lumbricoides, Ascaris suum or ‘Ascaris lumbrisuum’? The Journal of Infectious Diseases 213, 1355.Google Scholar
Dias da Silva, W and Tambourgi, DV (2010) IgY: a promising antibody for use in immunodiagnostic and in immunotherapy. Veterinary Immunology and Immunopathology 135, 173–180.Google Scholar
Dold, C and Holland, CV (2011) Ascaris and ascariasis. Microbes and Infection 13, 632–637.Google Scholar
Ferreira-Júnior, A, Santiago, FM, Silva, MV, et al. (2012) Production, characterization and applications for Toxoplasma gondii-specific polyclonal chicken egg yolk immunoglobulins. PLoS One 7, e40391.Google Scholar
Gassmann, M, Thömmes, P, Weiser, T and Hübscher, U (1990) Efficient production of chicken egg yolk antibodies against a conserved mammalian protein. FASEB Journal 4, 2528–2532.Google Scholar
Gonçalves, ALR, Silva, CV, Ueta, MT and Costa-Cruz, JM (2012) Antigen, antibody and immune complex detection in serum samples from rats experimentally infected with Strongyloides venezuelensis. Experimental Parasitology 130, 205–208.Google Scholar
Gonzaga, HT, Ribeiro, VS, Feliciano, ND, Manhani, MN, Silva, DA, Ueta, MT and Costa-Cruz, JM (2011) IgG avidity in differential serodiagnosis of human strongyloidiasis active infection. Immunology Letters 139, 87–92.Google Scholar
Gonzaga, HT, Vila-Verde, C, Nunes, DS, Ribeiro, VS, Cunha-Júnior, JP and Costa-Cruz, JM (2013) Ion-exchange protocol to obtain antigenic fractions with potential for serodiagnosis of strongyloidiasis. Parasitology 140, 69–75.Google Scholar
Kennedy, MW, Qureshi, F, Fraser, EM, Haswell-Elkins, MR, Elkins, DB and Smith, HV (1989) Antigenic relationships between the surface-exposed, secreted and somatic materials of the nematode parasites Ascaris lumbricoides, Ascaris suum, and Toxocara canis. Clinical and Experimental Immunology 75, 493–500.Google Scholar
Khurana, S and Sethi, S (2017) Laboratory diagnosis of soil transmitted helminthiasis. Tropical Parasitology 7, 86–91.Google Scholar
Khuroo, MS, Rather, AA, Khuroo, NS and Khuroo, MS (2016) Hepatobiliary and pancreatic ascariasis. World Journal of Gastroenterology 22, 7507–7517.Google Scholar
Kovacs-Nolan, J and Mine, Y (2012) Egg yolk antibodies for passive immunity. Annual Review of Food Science and Technology 3, 163–182.Google Scholar
Laemmli, UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.Google Scholar
Larsson, A, Carlander, D and Wilhelmsson, M (1998) Immune response in chicken with different amounts of antigen. Food and Agricultural Immunology 10, 29–36.Google Scholar
Lei, JH, Guan, F, Xu, H, Chen, L, Su, BT, Zhou, Y, Wang, T, Li, YL and Liu, WQ (2012) Application of an immunomagnetic bead ELISA based on IgY for detection of circulating antigen in urine of mice infected with Schistosoma japonicum. Veterinary Parasitology 187, 196–202.Google Scholar
Li, QV, Zhao, DH, Qu, HY and Zhou, CN (2014) Life-threatening complications of ascariasis in trauma patients: a review of the literature. World Journal of Emergency Medicine 5, 165–170.Google Scholar
Lowry, OH, Rosebrough, NJ, Farr, AL and Randall, RJ (1951) Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry 193, 265–275.Google Scholar
Lutz, A (1919) O Schistosoma mansoni e a schistosomose segundo observações feitas no Brasil. Memórias do Instituto Oswaldo Cruz 11, 121–125.Google Scholar
Manhani, MN, Ribeiro, VS, Cardoso, R, Ueira-Vieira, C, Goulart, LR and Costa-Cruz, JM (2011) Specific phage-displayed peptides discriminate different forms of neurocysticercosis by antibody detection in the serum samples. Parasite Immunology 33, 322–329.Google Scholar
Marzocchi-Machado, CM and Lucisano-Valim, YM (1997) Clearance de imunocomplexos: papel do complemento e dos polimorfonucleares neutrófilos. Medicina (Ribeirão Preto) 30, 234–242.Google Scholar
Moraes, RG (1948) Contribuição para o estudo do Strongyloides stercoralis e da estrongiloidíase no Brasil. Revista de Saúde Pública 1, 507–624.Google Scholar
Müller, S, Schubert, A, Zajac, J, Dyck, T and Oelkrug, C (2015) IgY antibodies in human nutrition for disease prevention. Nutrition Journal 14, 109–116.Google Scholar
Munhoz, LS, Vargas, GS, Fischer, G, Lima, M, Esteves, PA and Hübner, SO (2014) Avian IgY antibodies: characteristics and applications in immunodiagnostic. Ciência Rural 44, 153–160.Google Scholar
Nikolay, B, Brooker, SJ and Pullan, RL (2014) Sensitivity of diagnostic tests for human soil-transmitted helminth infections: a meta-analysis in the absence of a true gold standard. International Journal for Parasitology 44, 765–774.Google Scholar
Nogueira, DS, Gazzinelli-Guimarães, PH, Barbosa, FS, et al. (2016) Multiple exposures to Ascaris suum induce tissue injury and mixed Th2/Th17 immune response in mice. PLoS Neglected Tropical Diseases 10, e0004382.Google Scholar
Parija, SC, Chidambaram, M and Mandal, J (2017) Epidemiological and clinical features of soil-transmitted helminths. Tropical Parasitology 7, 81–85.Google Scholar
Pauly, D, Dorner, M, Zhang, X, Hlinak, A, Dorner, B and Schade, R (2009) Monitoring of laying capacity, immunoglobulin Y concentration, and antibody titer development in chickens immunized with ricin and botulinum toxins over a two-year period. Poultry Science 88, 281–290.Google Scholar
Pereira, EPV, van Tilburg, MF, Florean, EOPT and Guedes, MIF (2019) Egg yolk antibodies (IgY) and their applications in human and veterinary health: a review. International Immunopharmacology 73, 293–303.Google Scholar
Pullan, RL, Smith, JL, Jasrasaria, R and Brooker, SJ (2014) Global numbers of infection and disease burden of soil transmitted helminth infections in 2010. Parasites & Vectors 7, 37.Google Scholar
Salemi, S, Markovic, M, Martini, G and D'Amelio, R (2015) The expanding role of therapeutic antibodies. International Reviews of Immunology 34, 202–264.Google Scholar
Schwarzkopf, C, Staak, C, Behn, I and Erhard, M (2001) Immunisation. pp. 25–64 in Schade, R, Behn, I, Erhard, M, Hlinak, A and Staak, C (Eds) Chicken egg yolk antibodies, production and application. Berlin, IgY-Technology. Springer-Verlag.Google Scholar
Shimizu, M, Nagashima, H, Hashimoto, K and Suzuki, T (1994) Egg yolk antibody (IgY) stability in aqueous solution with high sugar concentrations. Journal of Food Science 59, 763–772.Google Scholar
Tini, M, Jewell, UR, Camenisch, G, Chilov, D and Gassmann, M (2002) Generation and application of chicken egg-yolk antibodies. Comparative Biochemistry and Physiology – Part A: Molecular and Integrative Physiology 131, 569–574.Google Scholar
Vilibic-Cavlek, T, Barbic, L, Stevanovic, V, Petrovic, G and Mlinaric-Galinovic, G (2016) IgG avidity: an important serologic marker for the diagnosis of tick-borne encephalitis virus infection. Polish Journal of Microbiology 65, 119–121.Google Scholar
Warr, GW, Magor, KE and Higgins, DA (1995) IgY: clues to the origins of modern antibodies. Immunology Today 16, 392–398.Google Scholar
Werkman, M, Wright, JE, Truscott, JE, et al. (2018) Testing for soil-transmitted helminth transmission elimination: Analysing the impact of the sensitivity of different diagnostic tools. PLoS Neglected Tropical Diseases 12, e0006114.Google Scholar
Yoshida, A, Kikuchi, T, Nakagaki, S and Maruyama, H (2016) Optimal ELISA antigen for the diagnosis of Ascaris suum infection in humans. Parasitology Research 115, 4701–4705.Google Scholar
Yoshihara, S, Ova, T, Furuya, T and Goto, N (1993) Use of body fluid of adult female Ascaris suum as an antigen in the enzyme-linked immunosorbent assay (ELISA) for diagnosis of swine ascariosis. Journal of Helminthology 67, 279–286.Google Scholar
Zhang, X, Calvert, RA, Sutton, BJ and Doré, KA (2017) IgY: a key isotype in antibody evolution. Biological Reviews of the Cambridge Philosophical Society 92, 2144–2156.Google Scholar