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Diagnostic Mr 31 000 Schistosoma mansoni proteins: requirement of infection, but not immunization, and use of the “miniblot” technique for the production of monoclonal antibodies

Published online by Cambridge University Press:  05 June 2009

Andreas Ruppel ,
Ute Breternitz  and
Reinhard Burger
Andreas Ruppel
Affiliation:
Institut für Tropenhygiene, Im Neuenheimer Feld 324, D–6900 Heidelberg, Federal Republic of Germany
Ute Breternitz
Affiliation:
Institut für Immunologie, Universitüt Heidelberg, Federal Republic of Germany
Reinhard Burger
Affiliation:
Institut für Immunologie, Universitüt Heidelberg, Federal Republic of Germany
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Abstract

Antibodies directed against diagnostic Mr 31 000 polypeptide(s) of adult Schistosoma mansoni were already formed in mice during prepatency. In contrast, repeated immunization of mice with homogenates of adult schistosomes failed to elicit antibodies detectable in immunoblots in the Mr 31 000 region. Therefore, spleen cells of infected mice were used to produce hybridoma lines. The “miniblot technique” was developed in order to detect in hybridoma supernatants antibodies against schistosome Mr 31 000 components. Electrophoretically separated total S. mansoni proteins were transferred onto nitrocellulose, and the position of the Mr 31 000 components was determined with polyclonal antisera and immunoblotting. Pieces of about 3 square mm of nitrocellulose bearing the diagnostic proteins were incubated with about 100 μ1 of hybridoma supernatant in microtitre plates and subsequently probed with peroxidase-conjugated antibody to mouse IgG. This screening technique identified hybridomas secreting antibody to the relevant S. mansoni antigens. It is applicable to other defined parasit antigens, which are, however, not available in biochemically purified form. The monoclonal antibodies produced against the proteins with diagnostic potential reacted with antigens localized in the schistosome gut.

Type
Research Papers
Information
Journal of Helminthology , Volume 61 , Issue 2 , June 1987 , pp. 95 - 101
Copyright
Copyright © Cambridge University Press 1987

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References

REFERENCES

Batteiger, B., Newhall, W. J. & Jones, R. B. (1982) The use of Tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes. Journal of Immunological Methods, 55, 297–307.CrossRefGoogle ScholarPubMed
Burger, R., Deubel, U., Hadding, U. & Bitter-Suermann, D. (1982) Identification of functionally relevant determinants on the complement component C3 with monoclonal antibodies. Journal of Immunology, 129, 2042–2050.CrossRefGoogle ScholarPubMed
Carlier, Y., Bout, D., Strecker, G., Debray, H. & Capron, A. (1980) Purification immunochemical, and biologic characterization of the Schistosoma circulating M antigen. Journal of Immunology, 124, 2442–2450.CrossRefGoogle ScholarPubMed
Chappell, C. L. & Dresden, M. H. (1986) Schistosoma mansoni: proteinase activity of “hemoglobinase” from the digestive tract of adult worms. Experimental Parasitology, 61, 160–167.CrossRefGoogle ScholarPubMed
Cordingley, J. S., Taylor, D. W., Dunne, D. W. & Butterworth, A. E. (1983) Clone banks of cDNA from the parasite Schistosoma mansoni: isolation of clones containing a potentially immunodiagnostic antigen gene. Gene, 26, 25–39.CrossRefGoogle ScholarPubMed
Deelder, A. M. & Kornelis, D. (1981) Immunodiagnosis of recently acquired Schistosoma mansoni infection. A comparison of various immunological techniques. Tropical and Geographical Medicine, 33, 36–41.Google ScholarPubMed
Deelder, A. M., Kornelis, D.Van Marck, E. A. E., Eveleigh, P. C. & Egmond, J. G. Van (1980) Schistosoma mansoni: Characterization of two circulating polysaccharide antigens and the immunological response to these antigens in Mouse, hamster and human infections. Experimental Parasitology, 50, 16–32.CrossRefGoogle ScholarPubMed
Deelder, A. M., Reinders, P. N. & Rotmans, J. P. (1977) Purification studies on an acidic protease from adult Schistosoma mansoni. Acta Leidensia, 45, 91–103.Google Scholar
Hawkes, R., Niday, E. & Gordon, J. (1982) Dot immunobinding assay for monoclonal and other antibodies. Analytic Biochemistry, 119, 142–147.CrossRefGoogle ScholarPubMed
Kearney, J. F., Radbruch, A.Liesegang, B. & Rajewski, K. (1979) A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. Journal of Immunology, 123, 1548–1550.CrossRefGoogle ScholarPubMed
Kelsoe, G. H. & Weller, T. H. (1978) Immunodiagnosis of infection with Schistosoma mansoni: Enzyme-linked immunosorbent assay for detection of antibody to circulating antigen. Proceedings of the National Academy of Sciences, 75, 5715–5717.CrossRefGoogle ScholarPubMed
Mitchell, G. F. & Cruise, K. M. (1986) Schistosomiasis: antigens and host-parasite interactions. In: Parasite antigens (Editor Pearson, T. W.) pp. 257–316. Marcel Dekker Inc: New York and Basel.Google Scholar
Mott, K. E. & Dixon, H. (1982) Collaborative study on antigens for immunodiagnosis of schistosomiasis. Bulletin of the World Health Organization, 60, 729–753.Google Scholar
Nash, T. E. (1978) Antibody response to a polysaccharide antigen present in the schistosome gut. I Sensitivity and specificity. American Journal of Tropical Medicine and Hygiene, 27, 938–943.Google Scholar
Nash, T. E. & Deelder, A. M. (1985) Comparison of four schistosome excretory-secretory antigens: Phenol sulfuric test active Peak, cathodic circulating antigen, gut-associated Proteoglycan, and circulating anodic antigen. American Journal of Tropical Medicine and Hygiene, 34, 236–241.CrossRefGoogle ScholarPubMed
Nash, T. E., Lunde, M. N. & Cheever, A. W. (1981) Analysis and antigenic activity of a carbohyrate fraction derived from adult Schistosoma mansoni. Journal of Immunology, 126, 805–810.CrossRefGoogle Scholar
Norden, A. P. & Strand, M. (1984) Schistosoma Mansoni S. haematobium, and S. japonicum: Identification of genus-, species-, and gender-specific antigenic worm glycoproteins. Experimental Parasitology, 57, 110–123.CrossRefGoogle ScholarPubMed
Oi, V. T. & Herzenberg, L. A. (1980) Immunoglobulin producing hybrid cell lines. In: Selected methods in cellular immunology (Editors Mishell, B. B. & Shigii, S. M.) 1st Edition, pp. 351–372. Freeman and Co.: San Francisco.Google Scholar
Rotmans, J. P. (1978) Schistosoma mansoni: Antigenic characterization of malate dehydrogenase isoenzymes and use in the defined antigen substrate spheres (DASS) system. Experimental Parasitology, 46, 49–58.CrossRefGoogle ScholarPubMed
Ruppel, A., Diesfeld, H. J., Hempelmann, E., Janitschke, K., Karam, M., Breternitz, U., Rother, U. & Burger, R. (1985 a) Schistosoma mansoni: Characterization of a protein with diagnostic potential using monospecific patient sera and monoclonal antibodies. Immunobiology, 170, 76.Google Scholar
Ruppel, A., Diesfeld, H. J. & Rother, U. (1985 b) Immunoblot analysis of Schistosoma mansoni antigens with sera of schistosomiasis patients: diagnostic potential of an adult schistosome polypeptide. Clinical and Experimental Immunology, 62, 499–506.Google Scholar
Ruppel, A., Rother, U. & Diesfeld, H. J. (1982) Schistosoma mansoni: Development of primary infections in mice genetically deficient or intact in the fifth component of complement. Parasitology, 85, 315–323.CrossRefGoogle ScholarPubMed
Ruppel, A., Rother, U., Vongerichten, H., Lucius, R. & Diesfeld, H. J. (1985c) Schistosoma mansoni: immunoblot analysis of adult worm proteins. Experimental Parasitology, 60, 195–206.CrossRefGoogle ScholarPubMed
Sauer, M. C. V. & Senft, A. W. (1972) Properties of a proteolytic enzyme from Schistosoma mansoni. Comparative Biochemistry and Parasitology, 42B, 205–220.Google Scholar
Senft, A. W., Goldberg, M. W. & Byram, J. B. (1981) Hemoglobinolytic activity of serum in mice infected with Schistosoma mansoni. American Journal of Tropical Medicine and Hygiene, 30, 96–101.CrossRefGoogle ScholarPubMed
Smithers, S. R. & Terry, R. J. (1965) The infection of laboratory hosts with cercariae of Schistosoma mansoni and the recovery of adult worms. Parasitology, 55, 695–700.CrossRefGoogle ScholarPubMed
Timms, A. R. & Bueding, E. (1959) Studies of a proteolytic enzyme from Schistosoma mansoni. British Journal of Pharmacology, 14, 68–73.Google ScholarPubMed