Hostname: page-component-cd9895bd7-hc48f Total loading time: 0 Render date: 2024-12-27T11:54:45.469Z Has data issue: false hasContentIssue false
  • English
  • Français

Characterization of fractionated Schistosoma mansoni soluble adult worm antigens that elicit human cell proliferation and granuloma formation in vitro

Published online by Cambridge University Press:  06 April 2009

C. Hirsch  and
A. M. Goes
C. Hirsch
Affiliation:
Departamento de Bioquímica e Imunologia, Institute de Ciências Biológicas, Universidade Federal de Minus Gerais, 30161-970 Belo Horizonte, MG, Brasil
A. M. Goes*
Affiliation:
Departamento de Bioquímica e Imunologia, Institute de Ciências Biológicas, Universidade Federal de Minus Gerais, 30161-970 Belo Horizonte, MG, Brasil
*
* Corresponding author. E-mail: goes@sagui.icb.ufmg.br.
Get access Rights & Permissions [Opens in a new window]

Summary

Soluble adult worm antigens (SWAP) of Schistosoma mansoni were fractionated by fast protein liquid chromatography (FPLC) system, using Q-Sepharose anion-exchange resin, in order to characterize antigenic fractions that may elicit cell responses in human schistosomiasis. SWAP fractions were eluted by 20 HIM Tris–HCl solution (pH 9·6) with an increasing gradient of 1 M NaCl. The FPLC system was able to resolve 6 fractions, enumerated I to VI, according to the NaCl gradient. The analysis of each fraction on SDS–PAGE showed that fractions I to IV were constituted by multiple protein bands with Mr ranging from 21 to > 200 kDa. Large amounts of nucleic acids were evidenced in fractions V and VI, as revealed by ethidium bromide staining of agarose electrophoresis gels. Using ELISA, it was shown that sera from chronic schistosomiasis patients contained antibodies that recognized antigens in practically all fractions. Studies were designed to investigate the capacity of these fractions to induce cell proliferation and granuloma formation. It was demonstrated that fraction III stimulated a significant proliferative response of peripheral blood mononuclear cells (PBMC) from chronic schistosomiasis patients. However, fraction III coupled to polyacrylamide beads induced small granuloma formation in vitro, whereas beads coated with fractions I, II and V were able to induce significant granuloma reactions.

Keywords

Schistosoma mansonigranulomaproliferationhuman cells
Type
Research Article
Information
Parasitology , Volume 112 , Issue 6 , June 1996 , pp. 529 - 535
Copyright
Copyright © Cambridge University Press 1996

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

Aitken, R., Coulson, P. S. & Wilson, R. A. (1988). Pulmonary leukocytic responses are linked to the acquired immunity of mice vaccinated with irradiated cercariae of Schistosoma mansoni. Journal of Immunology 140, 3573–9.CrossRefGoogle Scholar
Bahia-Oliveira, L. M., Gazzinelli, G., Eloi-Santos, S. M., Cunha-Melo, J. R., Alves-Oliveira, L. F., Silveira, A. M. S., Viana, I. R. C., Carmo, J., Souza, A. & Correia-Oliveira, R. (1992). Differential cellular reactivity to adult worm antigen of patients with clinical forms of schistosomiasis mansoni. Transactions of the Royal Society of Tropical Medicine and Hygiene 86, 57–61.CrossRefGoogle ScholarPubMed
Boros, D. L., Pelley, R. P. & Warren, K. S. (1975). Spontaneous modulation of granulomatous hypersensitivity in schistosomiasis mansoni. Journal of Immunology 114, 1437–41.CrossRefGoogle ScholarPubMed
Boros, D. L., Tomford, R. & Warren, K. S. (1977). Induction of granulomatous and elicitation of cutaneous sensitivity by partially purified SEA of Schistosomiasis mansoni. Journal of Immunology 118, 373–6.CrossRefGoogle Scholar
Bradford, M. M. (1976). A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–54.Google Scholar
Carter, C. E. & Colley, D. G. (1979). Partial purification of Schistosoma mansoni soluble egg antigen with Con A-sepharose chromatography. Journal of Immunology 122, 2204–9.CrossRefGoogle ScholarPubMed
Contigli, C., Doughty, B. L., Cone, J. C. & Goes, A. M. (1994). Recognition of different Schistosoma mansoni antigens by specific human T cell clones. Cellular Immunology 154, 77–87.CrossRefGoogle ScholarPubMed
Doughty, B. L. & Phillips, S. M. (1982). Delayed hypersensitivity granuloma formation around Schistosoma mansoni eggs in vitro. I. Definition of the model. Journal of Immunology 128, 30–6.Google Scholar
Gaafar, T., Ismail, S., Helmy, M., Afifi, A., Guirguis, N. & El Ridi, R. (1993). Identification of Schistosoma haematobium soluble egg antigens that elicit human granuloma formation in vitro. Parasitology Research 79, 103–8.Google Scholar
Goes, A. M., Gazzinelli, G., Rocha, R. S., Katz, N. & Doughty, B. L. (1991). Granulomatous hypersensitivity to Schistosoma mansoni egg antigens in human schistosomiasis. III. In vitro granuloma modulation induced by immune complexes. American Journal of Tropical Medicine and Hygiene 44, 434–43.CrossRefGoogle ScholarPubMed
Goes, A. M., Rocha, R. S., Gazzinelli, G. & Doughty, B. L. (1989). Production and characterization of human monoclonal antibodies against Schistosoma mansoni. Parasite Immunology 11, 695–711.CrossRefGoogle ScholarPubMed
Hamburger, J., Lustigman, S., Siongok, T. K. A., Ouma, J. H. & Mahmoud, A. A. F. (1982). Characterization of a purified glycoprotein from Schistosoma mansoni eggs: specificity, stability, and the involvement of carbohydrate and peptide moieties in serological activity. Journal of Immunology 128, 1864–9.Google Scholar
Harn, D. A., Danko, K., Quinn, J. J. & Stadecker, M. J. (1989). Schistosoma mansoni: the host immune response egg antigens. I. Partial characterization of cellular and humoral responses to pI fractions of soluble egg antigens. Journal of Immunology 142, 2061–6.CrossRefGoogle ScholarPubMed
Harn, D. A., Mitsuyama, M., Huguenel, E. D., Oligino, L. & David, J. R. (1985). Identification by monoclonal antibody of a major (28 kDa) surface membrane antigen of Schistosoma mansoni. Molecular and Biochemical Parasitology 16, 345–54.CrossRefGoogle Scholar
Hoffman, W. A., Pons, J. A. & Janer, J. L. (1934). The sedimentation–concentration method in schistosomiasis mansoni. Puerto Rico Journal of Public Health 9, 281–98.Google Scholar
Hsu, S. L., Hsu, H. F., Svestka, K. W. & Clark, W. (1986). Vaccination against schistosomiasis in mice with killed schistosomula without adjuvant. Proceedings of the Society of Experimental Biology and Medicine 181, 454–61.CrossRefGoogle ScholarPubMed
James, S. L., Deblois, L. A., Al-Zamel, F., Glaven, J. & Langhorne, J. (1986). Defective vaccinated-induced resistance to Schistosoma mansoni in P strain mice. III. Specificity of the associated defect in cell-mediated immunity. Journal of Immunology 137, 3959–67.Google Scholar
Kambara, T. & Wilson, R. A. (1990). In situ pulmonary responses of T cell and macrophage subpopulations to a challenge infection in mice vaccinated with irradiated cercariae of Schistosoma mansoni. Journal of Parasitology 76, 365–72.CrossRefGoogle ScholarPubMed
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–5.Google Scholar
Lukacs, N. W. & Boros, D. L. (1991). Splenic and granuloma T-lymphocyte responses to fractionated soluble egg antigens of Schistosoma mansoni infected mice. Infection and Immunity 59, 941–8.CrossRefGoogle ScholarPubMed
Lukacs, N. W. & Boros, D. L. (1992). Utilization of fractionated soluble egg antigens reveals selectively modulated granulomatous and lymphokine responses during murine schistosomiasis mansoni. Infection and Immunity 60, 3209–16.CrossRefGoogle ScholarPubMed
Lustigman, N. S., Mahmoud, A. A. F. & Hamburger, J. (1985). Glycopeptides in soluble egg antigen of Schistosoma mansoni: isolation, characterization and elucidation of their immunochemical and immunopathological relation to the major egg glycoprotein (MEG). Journal of Immunology 134, 1961–7.CrossRefGoogle Scholar
Payares, G., Kelly, C., Smithers, S. R. & Evans, W. H. (1985). Evidence that the 32, 38 and 20 K surface antigens of schistosomula and schistosomes are related proteins. Molecular and Biochemical Parasitology 17, 115–30.CrossRefGoogle Scholar
Phillips, S. M. & Colley, D. G. (1978). Immunologic aspects of host responses to schistosomiasis: resistance, immunopathology and eosinophil involvement. Progress in Allergy 24, 49–182.Google Scholar
Smith, M. A. & Clegg, J. A. (1985). Vaccination against Schistosoma mansoni with purified surface antigens. Science 277, 535–8.Google Scholar
Tiu, W. U., Davern, K. M., Wright, M. D., Board, P. G. & Mitchell, G. F. (1988). Molecular and serological characteristics of the glutathione S-transferases of Schistosoma japonicum and Schistosoma mansoni. Parasite Immunology 10, 693–706.CrossRefGoogle ScholarPubMed
Towbin, H. T., Staehelin, T. & Gordon, J. (1979). Electrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, USA 76, 4350–4.CrossRefGoogle ScholarPubMed
Vignali, D. A. A., Devery, M. E., Bickle, Q. D. & Taylor, M. G. (1990). The role of antibody affinity and litre in immunity to Schistosoma mansoni following vaccination with highly irradiated cercariae. Immunology 69, 195–201.Google Scholar
Warren, K. S. (1982). The secret of the immunopathogenesis of schistosomiasis: in vivo models. Immunological Review 61, 189–213.Google Scholar
Weiss, J. B., Aronstein, W. S. & Strand, M. (1987). Schistosoma mansoni: stimulation of artificial granuloma formation in vivo by carbohydrate determinants. Experimental Parasitology 64, 228–36.CrossRefGoogle ScholarPubMed
Wilkinson, L. (1989). Systat: the System for Statistic. Evanston, IL: Systat, Inc.Google Scholar
Wright, M. D., Melder, A. M., Daven, K. M. & Mitchell, G. P. (1991). Serologic reactivities of the 23-kDa integral membrane proteins of schistosomes. Journal of Immunology 147, 4338–42.Google Scholar