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Molecular cloning and characterization of a 21 kDa protein secreted from Trichinella pseudospiralis

Published online by Cambridge University Press:  12 April 2024

I. Nagano ,
Z. Wu ,
T. Nakada ,
A. Matsuo  and
Y. Takahashi
I. Nagano
Affiliation:
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan
Z. Wu
Affiliation:
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan
T. Nakada
Affiliation:
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan
A. Matsuo
Affiliation:
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan
Y. Takahashi*
Affiliation:
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan
*
*Author for correspondence Fax: 058 267 2960 E-mail: yu3@cc.gifu-u.ac.jp
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Abstract

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Recombinant protein was produced from the cDNA library of Trichinella pseudospiralis, which seemed to form part of the excretory–secretory (ES) products. The library was constructed from cDNA of muscle larvae at 1 month post-infection, and immunoscreened with antibody against T. pseudospiralis ES products. A clone, designated Tp21-3, contained a cDNA transcript of 657 bp in length with a single open reading frame, which encoded 172 amino acids (19617 Da in the estimated molecular mass). The predicted amino acid sequence of clone Tp21-3 had a similarity of 76% to that of clone ORF 17.20 (GenBank under accession number U88239) from T. spiralis. The recombinant fusion proteins encoded by clone Tp21-3 were produced in an Escherichia coli expression system and affinity purified. On Western blotting analysis, Tp21-3 recombinant proteins migrated at 40 kDa and reacted to antibody against T. pseudospiralis ES products and T. pseudospiralis-infected sera. Sera were developed against Tp 21-3 recombinant proteins, which reacted to a single band migrating at 21 kDa in crude worm extract and ES products from T. pseudospiralis on Western blotting analysis, and reacted with stichocytes of T. pseudospiralis on immunohistochemical staining.

Type
Research Article
Information
Journal of Helminthology , Volume 75 , Issue 3 , September 2001 , pp. 273 - 278
Copyright
Copyright © Cambridge University Press 2001

References

Alkarmi, T.O. & Faubert, G.M. (1981) Comparative analysis of mobility and ultrastructure of intramuscular larvae of Trichinella spiralis and Trichinella pseudospiralis . Journal of Parasitology 57, 685–691.Google Scholar
Almond, N.M., Mclaren, D.J. & Parkhouse, R.M.E. (1986) A comparison of the surface secretions of Trichinella pseudospiralis and T. spiralis . Parasitology 93, 163–176.CrossRefGoogle ScholarPubMed
Appleton, J.A., Bell, R.G.M., Homan, W. & Van, Knapen F. (1991) Consensus on Trichinella spiralis antigens and antibody. Parasitology Today 7, 190–192.CrossRefGoogle Scholar
Bove, S.N., Britovi, V.A. & Orluv, I.V. (1979) Species composition of Trichinella . Wiadomosci Parazytologiczne 25, 495–503.Google Scholar
Chung, Y.Y. & Ko, R.C. (1999) A novel cDNA clone encoding a specific excretory/secretory antigen of larval Trichinella pseudospiralis . Parasitology Research 85, 685–691.CrossRefGoogle ScholarPubMed
Denkers, E.Y., Hayes, C.E. & Wassom, D.L. (1991) Trichinella spiralis: influence of an immunodominant carbohydrate-associated determinant on the host antibody response. Experimental Parasitology 72, 403–410.CrossRefGoogle ScholarPubMed
Despommier, D.D. (1983) Biology pp. 31–73 in Campbell, W.C. (Ed) Trichinella and trichinosis . New York, Plenum Press.Google Scholar
Despommier, D.D., Gold, A.M., Buck, S.W., Capo, V. & Silberstein, D. (1990) Trichinella spiralis: secreted antigen of the infective L1 larva localizes to the cytoplasm and nucleoplasm of infected host cells. Experimental Parasitology 71, 27–38.CrossRefGoogle Scholar
Jasmer, D.P. (1990) Trichinella spiralis: altered expression of muscle proteins in trichinosis. Experimental Parasitology 70, 452–465.CrossRefGoogle ScholarPubMed
Kehayov, I., Tankov, C., Komandarev, S. & Kyurkchiev, S. (1991) Antigenic differences between Trichinella spiralis and T. pseudospiralis detected by monoclonal antibodies. Parasitology Research 77, 72–76.CrossRefGoogle Scholar
Ko, R.C. & Fan, L. (1996) Heat shock response of Trichinella spiralis and T. pseudospiralis . Parasitology 112, 89–95.CrossRefGoogle ScholarPubMed
Ko, R.C. & Mak, C.H. (1999) Trichinellosis as a model of new frontier research on parasitic infection. International Medical Research Journal 3, 21–31.Google Scholar
Ko, R.C., Fan, L., Lee, D.L. & Compton, H. (1994) Changes in host muscles induced by excretory/secretory products of larval Trichinella spiralis and Trichinella pseudospiralis . Parasitology 108, 195–203.CrossRefGoogle ScholarPubMed
Lee, D.L., Ko, R.C., Yi, X.Y. & Yeung, M.H.F. (1991) Trichinella spiralis: antigenic epitopes from the stichocytes detected in the hypertrophic nuclei and cytoplasm of the parasitized muscle fibre (nurse cell) of the host. Parasitology 102, 117–123.CrossRefGoogle ScholarPubMed
Mak, C.H. & Ko, R.C. (1999) Characterization of endonuclease activity from excretory/secretory products of a parasitic nematode, Trichinella spiralis . European Journal of Biochemistry 260, 477–481.CrossRefGoogle ScholarPubMed
Matsuo, A., Wu, Z., Nagano, I. & Takahashi, Y. (2000) Five types of nuclei present in the capsule of Trichinella spiralis . Parasitology 121, 203–210.CrossRefGoogle ScholarPubMed
Moczon, T. & Wranicz, M. (1999) Trichinella spiralis: proteinases in the larvae. Parasitology Research 85, 47–58.CrossRefGoogle ScholarPubMed
Nagano, I., Wu, Z., Nakada, T., Matsuo, A. & Takahashi, Y. (2001) Molecular cloning and characterization of a serine proteinase inhibitor from Trichinella spiralis Parasitology, in press.Google Scholar
Sambrook, J., Fritsch, E.F. & Maniatis, T. (1989) Screening expression libraries with antibodies and oligonucleotides Molecular cloning. A laboratory manual 2nd edn. pp. 12.1–12.44. New York, Cold Spring Harbor Laboratory Press.Google Scholar
Stewart, G.L., Wood, B.G. & Boley, R.B. (1985) Modulation of host response by Trichinella pseudospiralis . Parasite Immunology 7, 223–233.CrossRefGoogle ScholarPubMed
Stewart, G.L., Mann, M.A., Ubelaker, J.E., McCarthy, J.L. & Wood, B.G. (1988) A role for elevated plasma corticosterone in modulation of host response during infection with Trichinella pseudospiralis . Parasite Immunology 10, 139–150.CrossRefGoogle ScholarPubMed
Su, X.Z., Prestwood, A.K. & Mcgraw, R.A. (1991) Cloning and expression of complementary DNA encoding an antigen of Trichinella spiralis . Molecular and Biochemical Parasitology 45, 331–336.CrossRefGoogle ScholarPubMed
Todorova, V.K., Knox, D.P. & Kennedy, M.W. (1995) Proteinase in the excretory/secretory products (ES) of adult Trichinella spiralis . Parasitology 111, 201–208.CrossRefGoogle Scholar
Towbin, H., Staehelin, T. & Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy Sciences of the USA 76, 4350–4354.CrossRefGoogle ScholarPubMed
Vassilatis, D.K., Despommier, D.D., Misek, D.E., Polvere, R.I., Gold, A.M. & Van Der Ploeg, A.H.T. (1992) Analysis of a 43-kDa glycoprotein from the intracellular parasitic nematode Trichinella spiralis . Journal of Biological Chemistry 267, 18459–18465.CrossRefGoogle ScholarPubMed
Vassilatis, D.K., Despommier, D.D., Polvere, R.I., Gold, A.M. & Van Der Ploeg, A.H.T. (1996) Trichinella pseudospiralis secretes a protein related to the Trichinella spiralis 43-kDa glycoprotein. Molecular and Biochemical Parasitology 78, 25–31.CrossRefGoogle Scholar
Wakelin, D., Goyal, P.K., Dehlaw, M.S. & Hermanek, J. (1994) Immune responses to Trichinella spiralis and T. pseudospiralis in mice. Immunology 81, 475–479.Google Scholar
Wu, Z., Nagano, I. & Takahashi, Y. (1998) Differences and similarities between Trichinella spiralis and T. pseudospiralis in morphology of stichocyte granules, peptide maps of excretory and secretory (ES) products and messenger RNA of stichosomal glycoproteins. Parasitology 116, 61–66.CrossRefGoogle Scholar
Wu, Z., Nagano, I. & Takahashi, Y. (1999) A panel of antigens of muscle larvae of Trichinella spiralis and T. pseudospiralis as revealed by two-dimensional Western blot and immunoelectron microscopy. Parasitology 118, 615–622.CrossRefGoogle Scholar
Xu, D.,Wu, Z., Nagano, I. & Takahashi, Y. (1997) A muscle larva of Trichinella pseudospiralis is intracellular, but does not form a typical cyst wall. Parasitology International 46, 1–5.CrossRefGoogle Scholar
Yamasaki, H., Taib, R., Watanabe, Y., Mak, J.W., Zasmy, N., Araki, K., Chooi, L.P.K., Kita, K. & Aoki, T. (1998) Molecular characterization of a cDNA encoding an excretory-secretory antigen from Toxocara canis second stage larvae and its application to the immunodiagnosis of human toxocariasis. Parasitology International 47, 171–181.CrossRefGoogle Scholar
Zarlenga, D.S. & Gamble, H.R. (1990) Molecular cloning and expression of an immunodominant 53-Kda excretory-secretory antigen from Trichinella spiralis muscle larvae. Molecular and Biochemical Parasitology 42, 165–174.CrossRefGoogle ScholarPubMed
Zhang, Y.W., Lee, D.L. & Smith, J.E. (1993) Biochemical characterization of Trichinella spiralis and T. pseudospiralis stichocyte antigens. Applied Parasitology 34, 291–294.Google Scholar