Folia Parasitologica, vol. 54 (2007), issue 2

Folia Parasitologica 54[2] 94-98 (2007) | 10.14411/fp.2007.012

Peptidases of Trichobilharzia regenti (Schistosomatidae) and its molluscan host Radix peregra s. lat. (Lymnaeidae): construction and screening of cDNA library from intramolluscan stages of the parasite

Kateřina Dolečková1, Martin Kašný1, Libor Mikeš1, Francisca Mutapi2, Colin Stack3, Adrian P. Mountford4, Petr Horák1
1 Department of Parasitology, Faculty of Science, Charles University in Prague, Viničná 7, 128 44 Prague 2, Czech Republic
2 Institute for Infection and Immunology Research, University of Edinburgh, Ashworth Laboratories, King's Buildings, West Mains Rd., Edinburgh EH9 3JT, UK
3 Institute for the Biotechnology of Infectious Diseases, University of Technology, Sydney, P.O. Box 123, Broadway, NSW 2007 Sydney, Australia
4 Department of Biology (Area 5), The University of York, York YO10 5YW, UK

Trichobilharzia regenti is a neurotropic bird schistosome, causing cercarial dermatitis in humans. In this study, ZAP cDNA expression library from Radix peregra s. lat. hepatopancreases containing intramolluscan stages of T. regenti was constructed and screened using PCR with specific and degenerate primers, designed according to previously described serine and cysteine peptidases of other parasite species. Full-length sequences of cathepsins B1 and L, and two serine peptidases, named RpSP1 and RpSP2, were obtained. The protein-protein BLAST analysis and parallel control reactions with template from hepatopancreases of T. regenti non-infected snails revealed that only cathepsin B1 was of parasite origin. The remaining sequences were derived from the snail intermediate host, which implies that the initial source of parasite mRNA was contaminated by snail tissue. Regardless of this contamination, the cDNA library remains an excellent molecular tool for detection and identification of bioactive molecules in T. regenti cercariae.

Received: November 21, 2006; Accepted: January 30, 2007; Published: June 1, 2007


References

  1. BAHGAT M., FRANCKLOW K., DOENHOFF M.J., LI Y.L., RAMZY R.M., KIRSTEN C., RUPPEL A. 2001: Infection induces antibodies against the cercarial secretions, but not against the cercarial elastases of Schistosoma mansoni, Schistosoma haematobium, Schistosoma japonicum and Trichobilharzia ocellata. Parasite Immunol. 23: 557-565 Go to original source... Go to PubMed...
  2. BAHGAT M., RUPPEL A. 2002: Biochemical comparison of the serine protease (elastase) activities in cercarial secretions from Trichobilharzia ocellata and Schistosoma mansoni. Parasitol. Res. 88: 495-500 Go to original source... Go to PubMed...
  3. CAUMES E., FELDER-MOINET S., COUZIGOU C., DARRAS-JOLY C., LATOUR P., LEGER N. 2003: Failure of an ointment based on IR3535 (ethyl butylacetylaminopropionate) to prevent an outbreak of cercarial dermatitis during swimming races across Lake Annecy, France. Ann. Trop. Med. Parasitol. 97: 157- 163 Go to original source... Go to PubMed...
  4. CRISTOFOLETTI P.T., RIBEIRO A.F., TERRA W.R. 2005: The cathepsin L-like proteinases from the midgut of Tenebrio molitor larvae: sequence, properties, immunocytochemical localization and function. Insect Biochem. Mol. Biol. 35: 883-901 Go to original source... Go to PubMed...
  5. DALTON J.P., CLOUGH K.A., JONES M.K., BRINDLEY P.J. 1997: The cysteine proteinases of Schistosoma mansoni cercariae. Parasitology 114: 105-112 Go to original source... Go to PubMed...
  6. DE GENTILE L., PICOT H., BOURDEAU P., BARDET R., KERJAN A., PIRIOU M., LE GUENNIC A., BAYSSADE-DUFOUR C., CHABASSE D., MOTT K.E. 1996: Le dermatite cercarienne en Europe: un probleme de la sante publique nouveau? Bull. WHO 74: 159-163
  7. DVORAK J., DELCROIX M., ROSSI A., VOPALENSKY V., POSPISEK M., SEDINOVA M., MIKES L., SAJID M., SALI A., MCKERROW J.H., HORAK P., CAFFREY C.R. 2005: Multiple cathepsin B isoforms in schistosomula of Trichobilharzia regenti: identification, characterisation and putative role in migration and nutrition. Int. J. Parasitol. 35: 895-910 Go to original source... Go to PubMed...
  8. FAN J., MINCHELLA D.J., DAYS.R., MCMANUS D.P., TIU W.U., BRINDLEY P.J. 1998: Generation, identification, and evaluation of expressed sequence tags from different developmental stages of the Asian blood fluke Schistosoma japonicum. Biochem. Biophys. Res. Commun. 252: 348-356 Go to original source... Go to PubMed...
  9. FUNG M.C., LAUM.T., CHEN X.G. 2002: Expressed sequence tag (EST) analysis of a Schistosoma japonicum cercariae cDNA library. Acta Trop. 82: 215-224 Go to original source... Go to PubMed...
  10. HAAS W. 2003: Parasitic worms: strategies of host finding, recognition and invasion. Zoology 106: 349-364 Go to original source... Go to PubMed...
  11. HORAK P., KOLAROVA L., ADEMA C.M. 2002: Biology of the schistosome genus Trichobilharzia. Adv. Parasitol. 52: 155- 233 Go to original source... Go to PubMed...
  12. HORAK P., KOVAR L., KOLAROVA L., NEBESAROVA J. 1998: Cercaria-schistosomulum surface transformation of Trichobilharzia szidati and its putative immunological impact. Parasitology 116: 139-147 Go to original source... Go to PubMed...
  13. HU W., YAN Q., SHEN D.K., LIU F., ZHU Z.D., SONG H.D., XU X.R., WANG Z.J., RONG Y.P., ZENG L.C., WU J., ZHANG X., WANG J.J., XU X.N., WANG S.Y., FU G., ZHANG X.L., WANG Z.Q., BRINDLEY P.J., MCMANUS D.P., XUE C.L., FENG Z., CHEN Z., HAN Z.G. 2003: Evolutionary and biomedical implications of a Schistosoma japonicum complementary DNA resource. Nat. Genet. 35: 139-147 Go to original source... Go to PubMed...
  14. KOURILOVA P., HOGG K.G., KOLAROVA L., MOUNTFORD A.P. 2004: Cercarial dermatitis caused by bird schistosomes comprises both immediate and late phase cutaneous hypersensitivity reactions. J. Immunol. 172: 3766-3774 Go to original source... Go to PubMed...
  15. LARSEN A.H., BRESCIANI J., BUCHMANN K. 2004: Increasing frequency of cercarial dermatitis at higher latitudes. Acta Parasitol. 49: 217-221
  16. MEULEMAN E.A., HUYER A.R., LUUB T.W.J. 1984: Maintenance of the life cycle of Trichobilharzia ocellata via the duck Anas platyrhynchos and the pond snail Lymnaea stagnalis. Neth. J. Zool. 34: 414-417 Go to original source... Go to PubMed...
  17. MIKES L., ZIDKOVA L., KASNY M., DVORAK J., HORAK P. 2005: In vitro stimulation of penetration gland emptying by Trichobilharzia szidati and T. regenti (Schistosomatidae) cercariae. Quantitative collection and partial characterization of the products. Parasitol. Res. 96: 230-241 Go to original source... Go to PubMed...
  18. NEWPORT G.R., MCKERROW J.H., HEDSTROM R., PETITT M., MCGARRIGLE L., BARR P.J., AGABIAN N. 1988: Cloning of the proteinase that facilitates infection by schistosome parasites. J. Biol. Chem. 263: 13179-13184 Go to PubMed...
  19. PENG H.J., CHEN X.G., WANG X.Z., LUN Z.R. 2003: Analysis of the gene expression profile of Schistosoma japonicum cercariae by a strategy based on expressed sequence tags. Parasitol. Res. 90: 287-293 Go to original source... Go to PubMed...
  20. PRICE H.P., DOENHOFF M.J., SAYERS J.R. 1997: Cloning, heterologous expression and antigenicity of a schistosome cercarial protease. Parasitology 114: 447-453 Go to original source... Go to PubMed...
  21. RENARD G., GARCIA J.F., CARDOSO F.C., RICHTER M.F., SAKANARI J.A., OZAKI L.S., TERMINGNONI C., MASUDA A. 2000: Cloning and functional expression of a Boophilus microplus cathepsin L-like enzyme. Insect Biochem. Mol. Biol. 30: 1017-1026 Go to original source... Go to PubMed...
  22. RUDOLFOVA J., HAMPL V., BAYSSADE-DUFOUR C., LOCKYER A.E., LITTLEWOOD D.T., HORAK P. 2005: Validity reassessment of Trichobilharzia species using Lymnaea stagnalis as the intermediate host. Parasitol. Res. 95: 79-89 Go to original source... Go to PubMed...
  23. SAJID M., MCKERROW J.H., HANSELL E., MATHIEU M.A., LUCAS K.D., HSIEH I., GREENBAUM D., BOGYO M., SALTER J.P., LIM K.C., FRANKLIN C., KIM J.H., CAFFREY C.R. 2003: Functional expression and characterization of Schistosoma mansoni cathepsin B and its trans-activation by an endogenous asparaginyl endopeptidase. Mol. Biochem. Parasitol. 131: 65-75 Go to original source... Go to PubMed...
  24. SALTER J.P., CHOE Y., ALBRECHT H., FRANKLIN C., LIM K.C., CRAIK C.S., MCKERROW J.H. 2002: Cercarial elastase is encoded by a functionally conserved gene family across multi ple species of schistosomes. J. Biol. Chem. 277: 24618- 24624 Go to original source... Go to PubMed...
  25. SALTER J.P., LIM K.C., HANSELL E., HSIEH I., MCKERROW J.H. 2000: Schistosome invasion of human skin and degradation of dermal elastin are mediated by a single serine protease. J. Biol. Chem. 275: 38667-38673 Go to original source... Go to PubMed...
  26. SKELLY P.J., SHOEMAKER C.B. 2001: Schistosoma mansoni proteases Sm31 (cathepsin B) and Sm32 (legumain) are expressed in the cecum and protonephridia of cercariae. J. Parasitol. 87: 1218-1221 Go to original source... Go to PubMed...
  27. SKIRNISSON K., KOLAROVA L. 2005: [Swimmer's itch in Landmannalaugar, Iceland] Laeknabladid 91: 729-736. (In Icelandic.) Go to PubMed...