Hostname: page-component-586b7cd67f-2plfb Total loading time: 0 Render date: 2024-11-20T06:57:09.489Z Has data issue: false hasContentIssue false

Potential immunological markers for diagnosis of human strongyloidiasis using heterologous antigens

Published online by Cambridge University Press:  29 November 2016

M. A. CORRAL
Affiliation:
Laboratório de Investigação Médica (LIM-06), Hospital das Clínicas da Faculdade de Medicina, Universidade de São Paulo, SP, Brazil
F. M. PAULA
Affiliation:
Laboratório de Investigação Médica (LIM-06), Hospital das Clínicas da Faculdade de Medicina, Universidade de São Paulo, SP, Brazil Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo SP, Brazil
D. M. C. L. MEISEL
Affiliation:
Laboratório de Investigação Médica (LIM-06), Hospital das Clínicas da Faculdade de Medicina, Universidade de São Paulo, SP, Brazil
V. L. P. CASTILHO
Affiliation:
Divisão de Laboratório Central, Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brazil
E. M. N. GONÇALVES
Affiliation:
Divisão de Laboratório Central, Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brazil
D. LEVY
Affiliation:
Laboratório de Investigação Médica (LIM-31), Hospital das Clínicas da Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brazil
S. P. BYDLOWSKI
Affiliation:
Laboratório de Investigação Médica (LIM-31), Hospital das Clínicas da Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brazil
P. P. CHIEFFI
Affiliation:
Faculdade de Ciências Médicas, Santa Casa, São Paulo, SP, Brazil
W. CASTRO-BORGES
Affiliation:
Departamento de Ciências Biológicas, Universidade Federal de Ouro Preto, Ouro Preto, MG, Brazil
R. C. B. GRYSCHEK*
Affiliation:
Laboratório de Investigação Médica (LIM-06), Hospital das Clínicas da Faculdade de Medicina, Universidade de São Paulo, SP, Brazil Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo SP, Brazil
*
*Corresponding author: Laboratório de Investigação Médica (LIM-06), Hospital das Clínicas da Faculdade de Medicina, USP, São Paulo. Instituto de Medicina Tropical, prédio II, 2° andar, Av. Dr Enéas de Carvalho Aguiar, 470, Cerqueira Cesar, 05403-000, São Paulo, SP, Brazil. E-mail: [email protected]

Summary

Strongyloides venezuelensis is a parasitic nematode of rodents that is frequently used to obtain heterologous antigens for immunological diagnosis of human strongyloidiasis. The aim of this study was to identify antigens from filariform larvae of S. venezuelensis for immunodiagnosis of human strongyloidiasis. Soluble and membrane fractions from filariform larvae of S. venezuelensis were obtained in phosphate saline (SS and SM) and in Tris–HCl buffer (TS and TM), and were analysed by Western blotting. Different antigenic components were recognized by IgG antibodies from the sera of strongyloidiasis patients. Highest recognition was observed for a 30–40 kDa mass range present in all antigenic fractions. The band encompassing this mass range was then excised and subjected to mass spectrometry for protein identification. Immunoreactive proteins identified in the soluble fractions corresponded to metabolic enzymes, whereas cytoskeletal proteins and galectins were more abundant in the membrane fractions. These results represent the first approach towards identification of S. venezuelensis antigens for use in immunodiagnostic assays for human strongyloidiasis.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2016 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Anderson, N. W., Klein, D. M., Dornink, S. M., Jespersen, D. J., Kubofcik, J., Nutman, T. B., Merrigan, S. D., Couturier, M. R. and Theel, E. S. (2014). Comparison of three immunoassays for detection of antibodies to Strongyloides stercoralis . Clinical and Vaccine Immunology 21, 732736.CrossRefGoogle ScholarPubMed
Bahia, D., Avelar, L. G., Vigorosi, F., Cioli, D., Oliveira, G. C. and Mortara, R. A. (2006). The distribution of motor proteins in the muscles and flame cells of the Schistosoma mansoni miracidium and primary sporocyst. Parasitology 133(Pt 3), 321329.CrossRefGoogle ScholarPubMed
Bisoffi, Z., Buonfrate, D., Sequi, M., Mejia, R., Cimino, R. O., Krolewiecki, A. J., Albonico, M., Gobbo, M., Bonafini, S., Angheben, A., Requena-Mendez, A., Muñoz, J. and Nutman, T. B. (2014). Diagnostic accuracy of five serologic tests for Strongyloides stercoralis infection. PLoS Neglected Tropical Diseases 8, e2640.Google Scholar
Brindley, P. J., Gam, A. A., Pearce, E. J., Poindexter, R. W. and Neva, F. A. (1988). Antigens from the surface and excretions secretions of the filariform larva of Strongyloides stercoralis . Molecular and Biochemical Parasitology 28, 171180.Google Scholar
Buonfrate, D., Formenti, F., Perandin, F. and Bisoffi, Z. (2015). Novel approaches to the diagnosis of Strongyloides stercoralis infection. Clinical Microbiology and Infection 21, 543545.Google Scholar
Conway, D. J., Bailey, J. W., Lindo, J. F., Robinson, R. D., Bundy, D. A. P. and Bianco, A. E. (1993). Serum igg reactivity with 41-kda, 31-kda, and 28-kda larval proteins of Strongyloides stercoralis in individuals with strongyloidiasis. Journal of Infectious Diseases 168, 784787.CrossRefGoogle Scholar
Corral, M. A., Paula, F. M., Gottardi, M., Meisel, D. M. C. L., Castilho, V. L. P., Gonçalves, E. M. N., Chieffi, P. P. and Gryschek, R. C. B. (2015). Immunodiagnosis of human strongyloidiasis: use of six different antigenic fractions from Strongyloides venezuelensis parasitic females. Revista do Instituto de Medicina Tropical de Sao Paulo 57, 427430.Google Scholar
Feliciano, N. D., Gonzaga, H. T., Goncalves-Pires, M. D. R., Ribeiro-Goncalves, A. L., Rodrigues, R. M., Ueta, M. T. and Costa-Cruz, J. M. (2010). Hydrophobic fractions from Strongyloides venezuelensis for use in the human immunodiagnosis of strongyloidiasis. Diagnostic Microbiology and Infectious Disease 67, 153161.CrossRefGoogle ScholarPubMed
Fornelio, A. C., Gonzalez, A. J. and Caabeiro, F. R. (1995). Actin isoforms in the parasitic nematode Haemonchus contortus . Parasitology Research 81, 700702.Google Scholar
Garcia, L. S. (2001). Diagnostic Medical Parasitology, 4th Edn. American Society for Microbiology, Washington, DC.Google Scholar
Gonzaga, H. T., Ribeiro, V. S., Feliciano, N. D., Manhani, M. N., Silva, D. A. O., Ueta, M. T. and Costa-Cruz, J. M. (2011). IgG avidity in differential serodiagnosis of human strongyloidiasis active infection. Immunology Letters 139, 8792.Google Scholar
Greenhalgh, C. J., Loukas, A., Donald, D., Nikolaou, S. and Newton, S. E. (2000). A family of galectins from Haemonchus contortus . Molecular and Biochemical Parasitology 107, 117121.Google Scholar
Hunt, V. L., Tsai, I. J., Coghlan, A., Reid, A. J., Holroyd, N., Foth, B. J., Tracey, A., Cotton, J. A., Stanley, E. J., Beasley, H., Bennett, H. M., Brooks, K., Harsha, B., Kajitani, R., Kulkarni, A., Harbecke, D., Nagayasu, E., Nichol, S., Ogura, Y., Quail, M. A., Randle, N., Xia, D., Brattig, N. W., Soblik, H., Ribeiro, D. M., Sanchez-Flores, A., Hayashi, T., Itoh, T., Denver, D. R., Grant, W., Stoltzfus, J. D., Lok, J. B., Murayama, H., Wastling, J., Streit, A., Kikuchi, T., Viney, M. and Berriman, M. (2016). The genomic basis of parasitism in the Strongyloides clade of nematodes. Nature Genetics 48, 299307.Google Scholar
Kiel, M., Josh, P., Jones, A., Windon, R., Hunt, P. and Kongsuwan, K. (2007). Identification of immuno-reactive proteins from a sheep gastrointestinal nematode, Trichostrongylus colubriformis, using two-dimensional electrophoresis and mass spectrometry. International Journal for Parasitology 37, 14191429.Google Scholar
Koga, K., Kasuya, S., Khamboonruang, C., Sukhavat, K., Ieda, M., Takatsuka, N., Kita, K. and Ohtomo, H. (1991) A modified agar plate method for detection of Strongyloides stercoralis . American Journal of Tropical Medicine and Hygiene 45, 518521.Google Scholar
Laemmli, U. K. (1970). Cleavage of structural proteins during assembly of head of bacteriophage-T4. Nature 227, 680.CrossRefGoogle ScholarPubMed
Levenhagen, M. A. and Costa-Cruz, J. M. (2014). Update on immunologic and molecular diagnosis of human strongyloidiasis. Acta Tropica 135, 3343.Google Scholar
Lindo, J. F., Conway, D. J., Atkins, N. S., Bianco, A. E., Robinson, R. D. and Bundy, D. A. (1994). Prospective evaluation of enzyme-linked immunosorbent assay and immunoblot methods for the diagnosis of endemic Strongyloides stercoralis infection. American Journal of Tropical Medicine and Hygiene 51, 175179.Google Scholar
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J. (1951). Protein measurement with the folin phenol reagent. Journal of Biological Chemistry 193, 265275.Google Scholar
Machado, E. R., Faccioli, L. H., Costa-Cruz, J. M., Lourenco, E. V., Roque-Barreira, M. C., de Fatima Goncalves Pires, M. D. R. and Ueta, M. T. (2008). Strongyloides venezuelensis: the antigenic identity of eight strains for the immunodiagnosis of human strongyloidiasis. Experimental Parasitology 119, 714.Google Scholar
Marcilla, A., Sotillo, J., Perez-Garcia, A., Igual-Adell, R., Luz Valero, M., Sanchez-Pino, M. M., Bernal, D., Munoz-Antoli, C., Trelis, M., Toledo, R. and Esteban, J. G. (2010). Proteomic analysis of Strongyloides stercoralis L3 larvae. Parasitology 137, 15771583.Google Scholar
Newton, S. E., Monti, J. R., Greenhalgh, C. J., Ashman, K. and Meeusen, E. N. (1997). cDNA cloning of galectins from third stage larvae of the parasitic nematode Teladorsagia circumcincta . Molecular and Biochemical Parasitology 86, 143153.Google Scholar
Northern, C. and Grove, D. I. (1990). Strongyloides stercoralis antigenic analysis of infective larvae and adult worms. International Journal for Parasitology 20, 381387.Google Scholar
Olsen, A., van Lieshout, L., Marti, H., Polderman, T., Polman, K., Steinmann, P., Stothard, R., Thybo, S., Verweij, J. J. and Magnussen, P. (2009). Strongyloidiasis – the most neglected of the neglected tropical diseases? Transactions of the Royal Society of Tropical Medicine and Hygiene 103, 967972.Google Scholar
Pollard, T. D. and Borisy, G. G. (2003). Cellular motility driven by assembly and disassembly of actin filaments. Cell 112, 453–65.Google Scholar
Ravi, V., Ramachandran, S., Thompson, R. W., Andersen, J. F. and Neva, F. A. (2002). Characterization of a recombinant immunodiagnostic antigen (NIE) from Strongyloides stercoralis L3-stage larvae. Molecular and Biochemical Parasitology 125, 7381.CrossRefGoogle ScholarPubMed
Requena-Mendez, A., Chiodini, P., Bisoffi, Z., Buonfrate, D., Gotuzzo, E. and Munoz, J. (2013). The laboratory diagnosis and follow up of strongyloidiasis: a systematic review. PLoS Negl Trop Dis 7:e2002.Google Scholar
Rigo, C. R., Lescano, S. Z., de Marchi, C. R. and Amato Neto, V. (2008). Evaluation of the humoral immune response to Strongyloides venezuelensis antigens. Revista da Sociedade Brasileira de Medicina Tropical 41, 648653.Google Scholar
Rodrigues, R. M., Sopelete, M. C., Silva, D. A. D., Cunha-Junior, J. P., Taketomi, E. A. and Costa-Cruz, J. M. (2004). Strongyloides ratti antigenic antibodies in immunoblotting the immunodiagnosis components recognized by IgE as an additional tool for improving in human strongyloidiasis. Memorias do Instituto Oswaldo Cruz 99, 8993.CrossRefGoogle ScholarPubMed
Sato, Y., Inoue, F., Matsuyama, R. and Shiroma, Y. (1990). Immunoblot analysis of antibodies in human strongyloidiasis. Transactions of the Royal Society of Tropical Medicine and Hygiene 84, 403406.Google Scholar
Siddiqui, A. A., Koenig, N. M., Sinensky, M. and Berk, S. L. (1997). Strongyloides stercoralis: identification of antigens in natural human infections from endemic areas of the United States. Parasitology Research 83, 655658.Google Scholar
Silva, L. P., Barcelo, I. S. D., Passos-Lima, A. B., Espindola, F. S., Campos, D. M. B. and Costa-Cruz, J. M. (2003). Western blotting using Strongyloides ratti antigen for the detection of IgG antibodies as confirmatory test in human strongyloidiasis. Memorias do Instituto Oswaldo Cruz 98, 687691.CrossRefGoogle ScholarPubMed
Silva, C. V., Gonçalves, A. L., Cruz, L., Cruz, M. C., Ueta, M. T. and Costa-Cruz, J. M. (2013). F-actin accumulates in the vulva of female Strongyloides venezuelensis . Journal of Helminthology 87, 301304.CrossRefGoogle ScholarPubMed
Sudre, A. P., Siqueira, R. C., Barreto, M. G. M., Peralta, R. H. S., Macedo, H. W. and Peralta, J. M. (2007). Identification of a 26-kDa protein fraction as an important antigen for application in the immunodiagnosis of strongyloidiasis. Parasitology Research 101, 11171123.Google Scholar
Vasta, G. R. (2009). Roles of galectins in infection. Nature Reviews Microbiology 7, 424438.Google Scholar
Yan, F., Xu, L., Liu, L., Yan, R., Song, X. and Li, X. (2010). Immunoproteomic analysis of whole proteins from male and female adult Haemonchus contortus . Veterinary Journal 185, 174179.Google Scholar
Yan, R., Wang, J., Xu, L., Song, X. and Li, X. (2014). DNA vaccine encoding Haemonchus contortus Actin induces partial protection in Goats. Acta Parasitologica 59, 698709.Google Scholar
Young, A. R. and Meeusen, E. N. (2004). Galectins in parasite infection and allergic inflammation. Glycoconjugate Journal 19, 601606.CrossRefGoogle Scholar
Zhan, B., Ajmera, R., Geiger, S. M., Goncalves, M. T. P., Liu, Z., Wei, J., Wilkins, P. P., Fujiwara, R., Gazzinelli-Guimaraes, P. H., Bottazzi, M. B. and Hotez, P. (2015). Identification of immunodominant antigens for the laboratory diagnosis of toxocariasis. Tropical Medicine and International Health 20, 17871796.Google Scholar