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Analysis of cDNAs coding for immunologically dominant antigens from an oncosphere-specific cDNA library of Echinococcus multilocularis

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Abstract

A cDNA library based on mRNA from oncospheres of Echinococcus multilocularis was constructed and screened with an oncosphere-specific rabbit serum. cDNA sequences of three clones that were isolated out of this library are discussed: one codes for a serpin-like proteinase inhibitor, the first isolated from cestodes. Two other clones code for dominant oncosphere antigens and represent homologues of known genes: one is known from several taeniid cestodes as a protective antigen containing fibronectin III domains, the second is related to genes of small heat shock proteins. It contains an internal duplication that might be specific for platyhelminths.

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References

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1987) Current protocols in molecular biology. Greene Publishing Associates and Wiley Interscience, New York

  • Bairoch A (1992) PROSITE: a dictionary of sites and patterns in proteins. Nucleic Acids Res 20:2013–2018

    CAS  PubMed  Google Scholar 

  • Benitez L, Harrison LJS, Parkhouse RME, Garate T (1998) Sequence and preliminary characterisation of a Taenia saginata oncosphere gene homologue of the small heat-shock protein family. Parasitol Res 84:423–425

    Google Scholar 

  • Bilger B, Veit P, Müller V, Merckelbach A, Kersten D, Stöppler H, Lucius R (1995) Weitere Untersuchungen zum Befall des Rotfuchses mit Echinococcus multilocularis im Regierungsbezirk Tübingen. Tierärztl Umschau 50:465–470

    Google Scholar 

  • Blank CA, Brantly M (1994) Clinical features and molecular characteristics of α1-antitrypsin deficiency. Ann Allerg 72:105–121

    CAS  Google Scholar 

  • Blanton RE, Licate LS, Aman RA (1994) Characterization of a native and recombinant Schistosoma haematobium serine protease inhibitor gene product. Mol Biochem Parasitol 63:1–11

    Google Scholar 

  • Bork P, Doolittle RF (1993) Fibronectin type III modules in the receptor phosphatase CD45 and tapeworm antigen. Protein Sci 2:1185–1187

    CAS  PubMed  Google Scholar 

  • Byars NE, Allison AC (1987) Adjuvant formulation for use in vaccines to elicit both cell-mediated and humoral immunity. Vaccine 5:223–228

    Article  CAS  PubMed  Google Scholar 

  • Cai Y, Langley JG, Smith DI, Boros DL (1996) A cloned major Schistosoma mansoni egg antigen with homologies to small heat shock proteins elicits Th1 responsiveness. Infect Immun 64:1750–1755

    CAS  PubMed  Google Scholar 

  • Cao M, Chao H, Doughty BL (1993) Cloning of a cDNA encoding an egg antigen homologue from Schistosoma mansoni. Mol Biochem Parasitol 58:169–172

    Google Scholar 

  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299

    CAS  PubMed  Google Scholar 

  • Cohen FE, Gregoret LM, Amiri P, Aldape K, Railey J, McKerrow JH (1991) Arresting tissue invasion of a parasite by protease inhibitors chosen with the aid of computer modeling. Biochemistry 30:11221–11229

    CAS  PubMed  Google Scholar 

  • Curtis D, Lehmann R, Zamore PD (1995) Translational regulation in development. Cell 81:171–178

    CAS  PubMed  Google Scholar 

  • Dai WJ, Hemphill A, Waldvogel A, Ingold K, Deplazes P, Mossmann H, Gottstein B (2001) Major carbohydrate antigen of Echinococcus multilocularis induces an immunglobulin G response independent of αβ+ CD4+ T cells. Infect Immun 69:6074–6083

    CAS  PubMed  Google Scholar 

  • De Jong WW, Leunissen JAM, Voorter CEM (1993) Evolution of the α-crystallin/small heat-shock protein family. Mol Biol Evol 10:103–126

    PubMed  Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395

    PubMed  Google Scholar 

  • Eckert J, Deplazes P (1999) Alveolar echinococcosis in humans: the current situation in central Europe and the need for countermeasures. Parasitol Today 15:315–319

    Google Scholar 

  • Gaestel M, Vierling E, Buchner J (1999) The small heat shock protein (sHSP) family—an overview. In: Kreis T, Vale R (eds) Extracellular matrix, anchor, and adhesion proteins. Oxford University Press, Oxford, pp 269–272

  • Gauci CG, Lightowlers MW (1995) Developmental regulation of Taenia ovis 45 W gene expression. Mol Biochem Parasitol 73:263–266

    Google Scholar 

  • Gauci CG, Lightowlers MW (2001) Alternative splicing and sequence diversity of transcripts from the oncosphere stage of Taenia solium with homology to the 45 W antigen of Taenia ovis. Mol Biochem Parasitol 112:173–181

    Google Scholar 

  • Gauci C, Merli M, Müller V, Chow C, Yagi K, Mackenstedt U, Lightowlers MW (2002) Molecular cloning of a vaccine antigen against infection with the larval stage of Echinococcus multilocularis. Infect Immun 70:3969–3972

    Article  CAS  PubMed  Google Scholar 

  • Gerard GF, D'Alessio JM (1993) Reverse transcriptase (EC 2.7.7.49): the use of cloned MMLV-RT to synthesize DNA from RNA. Meth Mol Biol 16:73–93

    CAS  Google Scholar 

  • Gotoh O (1996) Significant improvements in accuracy of multiple protein sequence alignments by iterative refinement as assessed by reference to structural alignments. J Mol Biol 264:823–838

    Article  CAS  PubMed  Google Scholar 

  • Gottstein B, Hemphill A (1997) Immunpathology of echinococcosis. In: Freedman DO (ed) Immunopathogenetic aspects of disease induced by helminth parasites. Chem Immunol 66:177–208

    CAS  PubMed  Google Scholar 

  • Gottstein B, Saucy F, Deplazes P, Reichen J, Demierre G, Busato A, Zuercher C, Pugin P (2001) Is high prevalence of Echinococcus multilocularis in wild and domestic animals associated with disease incidence in humans? Emerg Infect Dis 7:408–412

    Google Scholar 

  • Gottstein B, Dai WJ, Walker M, Stettler M, Müller N, Hemphill A (2002) An intact laminated layer is important for the establishment of secondary Echinococcus multilocularis infection. Parasitol Res 88:822–828

    Google Scholar 

  • Huber R, Carrell RW (1989) Implications of the three-dimensional structure of α1-antitrypsin for structure and function of serpins. Biochemistry 28:8951–8966

    CAS  PubMed  Google Scholar 

  • Irving JA, Pike RN, Lesk AM, Whisstock JC (2000) Phylogeny of the serpin superfamily: implications of patterns of amino acid conservation for structure and function. Genome Res 10:1845–1864

    Article  CAS  PubMed  Google Scholar 

  • Ito A, Hashimoto A (1993) Vaccination with hatched but non-activated, non-viable oncospheres of Taenia taeniaeformis in rats. J Helminthol 67:165–168

    CAS  PubMed  Google Scholar 

  • Ito A, Asano K, Okamoto K-I (1994) Protective antibodies against Taenia taeniaeformis in rats infected with eggs or injected with non-viable oncospheres or recombinant antigens of oncospheres. Parasite Immunol 16:463–468

    Google Scholar 

  • Jeannin P, Lecoanet-Henchoz S, Delneste Y, Gauchat J-F, Bonnefoy J-Y (1998) Alpha-1 antitrypsin up-regulates human B cell differentiation selectively into IgE-and IgG4-secreting cells. Eur J Immunol 28:1815–1822

    Article  CAS  PubMed  Google Scholar 

  • Jones DT, Taylor WR, Thornton JM (1992) The rapid generation of mutation data matrices from protein sequences. Comput Appl Biosci 8:275–282

    CAS  PubMed  Google Scholar 

  • Krüger M, Osterburg G (1983) On the alignment of two or more molecular sequences. Comput Prog Biomed 16:68–69

    Google Scholar 

  • Langridge J, Langridge P, Bergquist PL (1980) Extraction of nucleic acids from agarose gels. Anal Biochem 103:264–271

    CAS  PubMed  Google Scholar 

  • Leboulle G, Rochez C, Louahed J, Rutti B, Brossard M, Bollen A, Godfroid E (2002) Isolation of Ixodes ricinus salivary gland mRNAs encoding factors induced during blood feeding. Am J Trop Med Hyg 66:225–233

    CAS  PubMed  Google Scholar 

  • Leid RW, Suquet CM, Bouwer HGA, Hinrichs DJ (1986) Interleukin inhibition by a parasite proteinase inhibitor, taeniaestatin. J Immunol 137:2700–2702

    CAS  PubMed  Google Scholar 

  • Leid RW, Grant RF, Suquet CM (1987) Inhibition of neutrophil aggregation by taeniaestatin, a cestode proteinase inhibitor. Int J Parasitol 17:1349–1353

    Google Scholar 

  • Li Z, King CL, Ogundipe JO, Licate LS, Blanton RE (1995) Preferential recognition by human IgE and IgG4 of a species-specific Schistosoma haematobium serine protease inhibitor. J Infect Dis 171:416–422

    CAS  PubMed  Google Scholar 

  • Lightowlers MW, Mitchell GF, Bowtell DDL, Anders RF, Rickard MD (1984) Immunization against Taenia taeniaeformis in mice: studies on the characterization of antigens from oncospheres. Int J Parasitol 14:321–333

    Google Scholar 

  • Lightowlers MW, Flisser A, Gauci CG, Heath DD, Jensen O, Rolfe R (2000) Vaccination against cysticercosis and hydatid disease. Parasitol Today 16:191–196

    Google Scholar 

  • Müller-Schollenberger V, Beyer W, Schnitzler P, Merckelbach A, Roth S, Kalinna BH, Lucius R (2001) Immunisation with Salmonella typhimurium-delivered glyceraldehyde-3-phosphate dehydrogenase protects mice against challenge infection with Echinococcus multilocularis eggs. Int J Parasitol 31:1441–1449

    Google Scholar 

  • Nene V, Dunne DW, Johnson KS, Taylor DW, Cordingley JS (1986) Sequence and expression of a major egg antigen from Schistosoma mansoni. Homologies to heat shock proteins and alpha-crystallins. Mol Biochem Parasitol 21:179–188

    Article  CAS  PubMed  Google Scholar 

  • Puissant C, Houdebine L-M (1990) An improvement of the single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. BioTechniques 8:148–149

    CAS  PubMed  Google Scholar 

  • Rajasekariah GR, Rickard MD, Mitchell GF (1980) Density-gradient separation of Taenia pisiformis oncospheres. J Parasitol 66:355–356

    Google Scholar 

  • Remold-O'Donnell E, Chin J, Alberts M (1992) Sequence and molecular characterization of human monocyte/neutrophil elastase inhibitor. Proc Natl Acad Sci U S A 89:5635–5639

    CAS  PubMed  Google Scholar 

  • Rogan MT, Craig PS, Zehyle E, Masinde G, Wen H, Zhou P (1992) In vitro killing of taeniid oncospheres, mediated by human sera from hydatid endemic areas. Acta Trop 51:291–296

    CAS  PubMed  Google Scholar 

  • Simon R, Richter JD (1994) Further analysis of cytoplasmic polyadenylation in Xenopus embryos and identification of embryonic cytoplasmic polyadenylation element-binding proteins. Mol Cell Biol 14:7867–7875

    CAS  PubMed  Google Scholar 

  • Sprecher CA, Morgenstern KA, Mathewes S, Dahlen JR, Schrader SK, Foster DC, Kisiel W (1995) Molecular cloning, expression, and partial characterization of two novel members of the ovalbumin family of serine proteinase inhibitors. J Biol Chem 270:29854–29861

    Article  CAS  PubMed  Google Scholar 

  • Strimmer K, von Haeseler A (1996) Quartet puzzling: a quartet maximum-likelihood method for reconstructing tree topologies. Mol Biol Evol 13:964–969

    CAS  Google Scholar 

  • Sun J, Rose JB, Bird P (1995) Gene structure, chromosomal localization, and expression of the murine homologue of human proteinase inhibitor 6 (PI-6) suggests divergence of PI-6 from the ovalbumin serpins. J Biol Chem 270:16089–16096

    Article  CAS  PubMed  Google Scholar 

  • Suquet C, Green Edwards C, Leid RW (1984) Isolation and partial characterization of a Taenia taeniaeformis metacestode proteinase inhibitor. Int J Parasitol 14:165–172

    Google Scholar 

  • Veit P, Bilger B, Schad V, Schäfer J, Frank W, Lucius R (1995) Influence of environmental factors on the infectivity of Echinococcus multilocularis eggs. Parasitology 110:79–86

    Google Scholar 

  • Zorio DAR, Cheng NN, Blumenthal T, Spieth J (1994) Operons as a common form of chromosomal organization in C. elegans. Nature 372:270–272

    CAS  PubMed  Google Scholar 

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Acknowledgements

We gratefully acknowledge the help of D. Kersten and H. Stöppler (Aulendorf) and U. Eskens (Gießen), as well as their assistants in collecting fox intestines. Many thanks to B. Bilger and V. Müller-Schollenberger for their help in the inspection of this material and for providing anti-metacestode serum and the GAPDH probe; to B. Kübler for technical assistance, to U. Schneider (Aachen) for financial support, to S. Meuer and A. Ruppel (Heidelberg) for disposition of infrastructure for DNA sequencing and completion of this manuscript. We thank G. Roger, A. Ruppel and U. Vettel for critically reading the manuscript. The project was supported by a DFG-grant to R. Lucius (Lu 325/4-5); by U. Schneider (Orthopädische Universitätsklinik Aachen) and by the Arbeitsamt Mannheim. The experiments described in this publication have been done in compliance with German legislation. The DNA sequences discussed in this paper have been submitted to the EMBL data bank under the following accession numbers: aj420235—ONCO1, aj420236—ONCO2, aj420237—SERP; the corresponding derived amino acid sequences are available in the SPTREMBL data base under the respective accession numbers: q8wpi6, q8wt42, q8wt41.

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Authors and Affiliations

  1. Department of Parasitology, Institute of Zoology, University of Hohenheim, 70599, Stuttgart, Germany

    Armin Merckelbach, Martina Wager & Richard Lucius

  2. Department of Tropical Hygiene, University of Heidelberg, Im Neuenheimer Feld 324, 69120, Heidelberg, Germany

    Armin Merckelbach

  3. Department of Molecular Parasitology (Institute of Biology), Humboldt University, Philippstrasse 13, 10115 , Berlin, Germany

    Richard Lucius

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  1. Armin Merckelbach
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Correspondence to Armin Merckelbach.

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Merckelbach, A., Wager, M. & Lucius, R. Analysis of cDNAs coding for immunologically dominant antigens from an oncosphere-specific cDNA library of Echinococcus multilocularis . Parasitol Res 90, 493–501 (2003). https://doi.org/10.1007/s00436-003-0888-4

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  • DOI: https://doi.org/10.1007/s00436-003-0888-4

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