Schistosoma mansoni: identification of a 46KDa antigen of the schistosomular surface by monoclonal antibody

Toledo, V. P. C. P.; Tavares, C. A. P.; Henriques, S. B.; Nascimento, E.; Resende, M.

doi:10.1590/S0036-46651994000300002

Abstracts

An IgG2a subclass monoclonal antibody, C6G9, was obtained by immunization of BALB/c mice with Schistosoma mansoni egg antigens. With this monoclonal antibody, it was possible to identify a schistosomular antigen with a molecular weight of 46 kilodaltons (KDa), and its expression being evaluated by means of indirect immunofluorescence. The antigen persisted in the integument of the developing schistosomulum, for at least 96 hours post-transformation. The monoclonal antibody also reacted with the cercaria surface, but not with that of adult worm. The C6G9 was also able to mediate significant levels of cytotoxicity in the presence of complement for newly transformed schistosomula.

Schistosoma mansoni; Monoclonal antibody; Cercariae; Schistosomula

Um anticorpo monoclonal da subclasse IgG2a, designado C6G9, foi obtido pela imunização de camundongos BALB/c com antígenos de ovo de Schistosoma mansoni. Esse anticorpo monoclonal possibilitou a identificação de um antigeno de peso molecular aproximado de 46 quilodaltons (KDa), cuja expressão foi avaliada através da reação de imunofluorescência indireta. O referido antígeno persistiu no tegumento do esquistossômulo em desenvolvimento pelo menos até 96 horas pós-transformação. O anticorpo monoclonal reagiu também com a superfície de cercárias, mas não com a de vermes adultos. O C6G9, em presença de complemento, foi também capaz de mediar níveis significativos de citoxicidade para esquistossômulos recém-transformados.

SEROLOGY

Schistosoma mansoni: identification of a 46KDa antigen of the schistosomular surface by monoclonal antibody

Schistosoma mansoni: identificação de um antígeno de 46KDa da superfície de esquistossômulo por anticorpo monoclonal

V. P. C. P. Toledo^I; C. A. P. Tavares^II; S. B. Henriques^II; E. Nascimento^I; M. Resende^III

^IDepartamento de Parasitologia do Instituto de Ciências Biológicas da UFMG

^IIDepartamento de Bioquímica e Imunologia do Instituto de Ciências Biológicas da UFMG

^IIIDepartamento de Microbiologia do Instituto de Ciências Biológicas da UFMG

^{Address for correspondence} Address for correspondence: Vicente de Paulo C. P. Toledo Departamento de Parasitologia, ICB/UFMG Cx.P. 486 CEP 30161-970 Belo Horizonte, MG, Brasil

SUMMARY

An IgG_2a subclass monoclonal antibody, C₆G₉, was obtained by immunization of BALB/c mice with Schistosoma mansoni egg antigens. With this monoclonal antibody, it was possible to identify a schistosomular antigen with a molecular weight of 46 kilodaltons (KDa), and its expression being evaluated by means of indirect immunofluorescence. The antigen persisted in the integument of the developing schistosomulum, for at least 96 hours post-transformation. The monoclonal antibody also reacted with the cercaria surface, but not with that of adult worm. The C₆G₉ was also able to mediate significant levels of cytotoxicity in the presence of complement for newly transformed schistosomula.

Keywords:Schistosoma mansoni; Monoclonal antibody, Cercariae; Schistosomula.

RESUMO

Um anticorpo monoclonal da subclasse IgG_2a, designado C₆G₉, foi obtido pela imunização de camundongos BALB/c com antígenos de ovo de Schistosoma mansoni. Esse anticorpo monoclonal possibilitou a identificação de um antigeno de peso molecular aproximado de 46 quilodaltons (KDa), cuja expressão foi avaliada através da reação de imunofluorescência indireta. O referido antígeno persistiu no tegumento do esquistossômulo em desenvolvimento pelo menos até 96 horas pós-transformação. O anticorpo monoclonal reagiu também com a superfície de cercárias, mas não com a de vermes adultos. O C₆G₉, em presença de complemento, foi também capaz de mediar níveis significativos de citoxicidade para esquistossômulos recém-transformados.

Full text available only in PDF format.

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ACKNOWLEDGEMENT

We are indebted to Dr. Carlos Alberto da Costa and Dr. Paul Williams for the critical analysis of this paper.

Recebido para publicação em 30/09/1993.

Aceito para publicação em 04/02/1994.

1. ARONSTEIN, W.S.; NORDEN, A.P. & STRAND, M. - Tegumental expression in larval and adult stages of a major schistosome structural glycoprotein. Amer. J. trop. Med. Hyg., 32: 334-342, 1983.
2. BENNETT, L.J. & SEED, J.L. - Characterization and isolation of Concanavalin A binding sites from the epidermis of Schistosoma mansoni. J. Parasit, 63: 250-258, 1977.
3. BRADFORD, M.M. - A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt. Biochem., 72: 248-254, 1976.
4. CHEN, L.; REKOSH, D.M. & LoVERDE, P.T. - Schistosoma mansoni p48 eggshell protein gene: characterization, develop-mentally regulated expression and comparison to the p14 eggshell protein gene. Molec. Biochem. Parasit., 52: 39-52, 1992.
5. DISSOUS, C.; GRZYCH, J.M. & CAPRON, A. - Schistosoma mansoni surface antigen defined by a rat monoclonal IgG_2a J. Immunol., 129: 2232-2234, 1982.
6. FREITAS, J.R.; RESENDE, E.S.; JUNQUEIRA, D.V.; COSTA, A.M. & PELLEGRINO, J. - Criaçăo em massa e ritmo de crescimento da Biomphalaria glabrata. Cienc. e Cult., 23: 968-974, 1975.
7. GALFRE, G. & MILSTEIN, C. - Preparation of monoclonal antibodies:strategies and procedures. In: SYMPOSIUM OF THE PROPERTIES OF THE MONOCLONAL ANTIBODIES PRODUCED BY HYBRIDOMA TECHNOLOGY AND THEIR APPLICATION TO THE STUDY OF DISEASES, Singapore, 1981. Geneva, UNDP/WORLD BANK/WHO, 1982. Proceedings. p. 1-46.
8. GODDING, J.W. - Conjugation of antibodies with fluorochromes: modifications to the standard methods J. immunol. Meth., 13: 215-226, 1976.
9. HARN, D.; MITSUYAMA, M. & DAVID, J.R. - Schistosoma mansoni: anti-egg monoclonal antibodies protect against cercarial challenge in vivo. J. exp. Med., 159: 1371-1387, 1988.
10. HAYUNGA, E.G.; MURRELL, K.D.; TAYLOR, D.W. & VANNIER, W.E. - Isolation and characterization of surface antigens from Schistosoma mansoni. II. Antigenicity of radiolabelled proteins from adult worms. J. Parasit., 65: 497-506, 1979.
11. OMER-ALI, P.; HAGAN, F.; HACKETT, F.; SMITHERS, S.R. & SIMPSON, A.J.G. - Variable species and stage specificity of schistosomulum surface epitopes recognized by mice vaccinated with highly irradiated cercariae. Paras. Immunol., 11: 257-267, 1989.
12. OUCHTERLONY, O. & NILSSON, L.A. - Immunodiffusion and immunoelectrophoresis. In: WEIR, D. M. - Handbook of experimental immunology. 2 ed. Oxford, Blackwell, 1973. v. 1, p. 19. 1-19.39.
13. PAYARES, G.; McLAREN, D.J.; EVANS, W.H. & SMITHERS, S.R. - Changes in the surface antigen profile of Schistosoma mansoni during maturation from cercariae to adult worm. Parasitology, 91: 83-99, 1985.
14. RAMALHO-PINTO, F.J.; GAZZINELLI, G.; HOWELLS,R.E. et al. - Schistosoma mansoni: defined system for step-wise transformation of cercariae to schistosomula in vitro. Exp. Parasit., 36: 360-372, 1974.
15. ROBERTS, S.M.; AITKEN, R.; VOJVODIC, M.; WELLS, E. & WILSON, R.A. - Identification of exposed components on the surface of adult Schistosoma mansoni by lactoperoxidase-catalysed iodination. Molec. Biochem. Parasit., 9: 129-143, 1983.
16. RUMJANECK, F.D.; McLAREN, D.J. & SMITHERS, S.R. - Serum induced expression of a surface proteins in schistosomula of Schistosoma mansoni: a possible receptor for a lipid uptake. Molec. Biochem. Parasit., 9: 337-349, 1983.
17. SIMPSON, A.J.G. - Schistosome surface antigens: developmental expression and immunological function. Parasit. today, 6: 40-45, 1990.
18. SIMPSON, A.J.G.; JAMES, S.L. & SHER, A. - Identification of surface antigens of schistosomula of Schistosoma mansoni recognized by chronic infection and by exposure to highly irradiated cercariae. Infect. Immun., 41: 591-597, 1983.
19. SIMPSON, A.J.G.; PAYARES, G.; WALKER, T.; KNIGHT, M. & SMITHERS, S.R. - The modulation of expression of polypeptide surface antigens on developing schistosomula of Schistosoma mansoni. J. Immunol., 133: 2725-2730, 1984.
20. SMITH, A.L., HUGGINS, M.C. & HAVERCROFT, C. - Schistosoma mansoni: adult males and females differentially express antigens encoded by repetitive genomic DNA. Parasitology, 105: 63-69, 1992.
21. SMITH, M.A. & CLEGG, J.A. - Vaccination against Schistosoma mansoni with purified surface antigens. Science, 227: 535-538, 1985.
22. SMITH, M.A.; CLEGG, J.A. & SNARY, D. - Monoclonal antibodies demonstrate similarity of surface antigens on different clones of Schistosoma mansoni schistosomula. Trans. roy. Soc. trop. Med. Hyg., 78: 187-189, 1984.
23. SMITHERS, S.R.; TERRY, R.J. & HOCKLEY, D.J. - Host antigens in schistosomiasis. Proc. roy. Soc. B., 171: 483-494, 1969.
24. TAVARES, C.A.P.; DE ROSSI, R.; PAYARES, G. et al. - A monoclonal antibody raised against adult Schistosoma mansoni which recognizes a surface antigen on schistosomula. Z. Parasitenk., 70: 189-197, 1984.
25. TAYLOR, D.W. & BUTTERWORTH, A.E. - Monoclonal antibodies against surface antigens of schistosomula of Schistosoma mansoni. Parasitology, 84: 65-82, 1982.
26. ZODDA, D.M. & PHILLIPS, S.M. - Monoclonal antibody mediated protection against Schistosoma mansoni infection in mice. J. Immunol., 129: 2326-2328, 1982.

Address for correspondence:

Vicente de Paulo C. P. Toledo

Departamento de Parasitologia, ICB/UFMG

Cx.P. 486

CEP 30161-970 Belo Horizonte, MG, Brasil

Publication Dates

Publication in this collection
20 Sept 2006
Date of issue
June 1994

History

Accepted
04 Feb 1994
Received
30 Sept 1993

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. ARONSTEIN, W.S.; NORDEN, A.P. & STRAND, M. - Tegumental expression in larval and adult stages of a major schistosome structural glycoprotein. Amer. J. trop. Med. Hyg., 32: 334-342, 1983.

[2] 2. BENNETT, L.J. & SEED, J.L. - Characterization and isolation of Concanavalin A binding sites from the epidermis of Schistosoma mansoni. J. Parasit, 63: 250-258, 1977.

[3] 3. BRADFORD, M.M. - A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt. Biochem., 72: 248-254, 1976.

[4] 4. CHEN, L.; REKOSH, D.M. & LoVERDE, P.T. - Schistosoma mansoni p48 eggshell protein gene: characterization, develop-mentally regulated expression and comparison to the p14 eggshell protein gene. Molec. Biochem. Parasit., 52: 39-52, 1992.

[5] 5. DISSOUS, C.; GRZYCH, J.M. & CAPRON, A. - Schistosoma mansoni surface antigen defined by a rat monoclonal IgG_2a J. Immunol., 129: 2232-2234, 1982.

[6] 6. FREITAS, J.R.; RESENDE, E.S.; JUNQUEIRA, D.V.; COSTA, A.M. & PELLEGRINO, J. - Criaçăo em massa e ritmo de crescimento da Biomphalaria glabrata. Cienc. e Cult., 23: 968-974, 1975.

[7] 7. GALFRE, G. & MILSTEIN, C. - Preparation of monoclonal antibodies:strategies and procedures. In: SYMPOSIUM OF THE PROPERTIES OF THE MONOCLONAL ANTIBODIES PRODUCED BY HYBRIDOMA TECHNOLOGY AND THEIR APPLICATION TO THE STUDY OF DISEASES, Singapore, 1981. Geneva, UNDP/WORLD BANK/WHO, 1982. Proceedings. p. 1-46.

[8] 8. GODDING, J.W. - Conjugation of antibodies with fluorochromes: modifications to the standard methods J. immunol. Meth., 13: 215-226, 1976.

[9] 9. HARN, D.; MITSUYAMA, M. & DAVID, J.R. - Schistosoma mansoni: anti-egg monoclonal antibodies protect against cercarial challenge in vivo. J. exp. Med., 159: 1371-1387, 1988.

[10] 10. HAYUNGA, E.G.; MURRELL, K.D.; TAYLOR, D.W. & VANNIER, W.E. - Isolation and characterization of surface antigens from Schistosoma mansoni. II. Antigenicity of radiolabelled proteins from adult worms. J. Parasit., 65: 497-506, 1979.

[11] 11. OMER-ALI, P.; HAGAN, F.; HACKETT, F.; SMITHERS, S.R. & SIMPSON, A.J.G. - Variable species and stage specificity of schistosomulum surface epitopes recognized by mice vaccinated with highly irradiated cercariae. Paras. Immunol., 11: 257-267, 1989.

[12] 12. OUCHTERLONY, O. & NILSSON, L.A. - Immunodiffusion and immunoelectrophoresis. In: WEIR, D. M. - Handbook of experimental immunology. 2 ed. Oxford, Blackwell, 1973. v. 1, p. 19. 1-19.39.

[13] 13. PAYARES, G.; McLAREN, D.J.; EVANS, W.H. & SMITHERS, S.R. - Changes in the surface antigen profile of Schistosoma mansoni during maturation from cercariae to adult worm. Parasitology, 91: 83-99, 1985.

[14] 14. RAMALHO-PINTO, F.J.; GAZZINELLI, G.; HOWELLS,R.E. et al. - Schistosoma mansoni: defined system for step-wise transformation of cercariae to schistosomula in vitro. Exp. Parasit., 36: 360-372, 1974.

[15] 15. ROBERTS, S.M.; AITKEN, R.; VOJVODIC, M.; WELLS, E. & WILSON, R.A. - Identification of exposed components on the surface of adult Schistosoma mansoni by lactoperoxidase-catalysed iodination. Molec. Biochem. Parasit., 9: 129-143, 1983.

[16] 16. RUMJANECK, F.D.; McLAREN, D.J. & SMITHERS, S.R. - Serum induced expression of a surface proteins in schistosomula of Schistosoma mansoni: a possible receptor for a lipid uptake. Molec. Biochem. Parasit., 9: 337-349, 1983.

[17] 17. SIMPSON, A.J.G. - Schistosome surface antigens: developmental expression and immunological function. Parasit. today, 6: 40-45, 1990.

[18] 18. SIMPSON, A.J.G.; JAMES, S.L. & SHER, A. - Identification of surface antigens of schistosomula of Schistosoma mansoni recognized by chronic infection and by exposure to highly irradiated cercariae. Infect. Immun., 41: 591-597, 1983.

[19] 19. SIMPSON, A.J.G.; PAYARES, G.; WALKER, T.; KNIGHT, M. & SMITHERS, S.R. - The modulation of expression of polypeptide surface antigens on developing schistosomula of Schistosoma mansoni. J. Immunol., 133: 2725-2730, 1984.

[20] 20. SMITH, A.L., HUGGINS, M.C. & HAVERCROFT, C. - Schistosoma mansoni: adult males and females differentially express antigens encoded by repetitive genomic DNA. Parasitology, 105: 63-69, 1992.

[21] 21. SMITH, M.A. & CLEGG, J.A. - Vaccination against Schistosoma mansoni with purified surface antigens. Science, 227: 535-538, 1985.

[22] 22. SMITH, M.A.; CLEGG, J.A. & SNARY, D. - Monoclonal antibodies demonstrate similarity of surface antigens on different clones of Schistosoma mansoni schistosomula. Trans. roy. Soc. trop. Med. Hyg., 78: 187-189, 1984.

[23] 23. SMITHERS, S.R.; TERRY, R.J. & HOCKLEY, D.J. - Host antigens in schistosomiasis. Proc. roy. Soc. B., 171: 483-494, 1969.

[24] 24. TAVARES, C.A.P.; DE ROSSI, R.; PAYARES, G. et al. - A monoclonal antibody raised against adult Schistosoma mansoni which recognizes a surface antigen on schistosomula. Z. Parasitenk., 70: 189-197, 1984.

[25] 25. TAYLOR, D.W. & BUTTERWORTH, A.E. - Monoclonal antibodies against surface antigens of schistosomula of Schistosoma mansoni. Parasitology, 84: 65-82, 1982.

[26] 26. ZODDA, D.M. & PHILLIPS, S.M. - Monoclonal antibody mediated protection against Schistosoma mansoni infection in mice. J. Immunol., 129: 2326-2328, 1982.

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