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Immunoblot analysis of serum IgG, IgA and IgE responses against larval excretory-secretory antigens of Anisakis simplex in patients with gastric anisakiasis

Published online by Cambridge University Press:  05 June 2009

Nobuaki Akao ,
Taka-Aki Ohyama  and
Kaoru Kondo
Nobuaki Akao
Affiliation:
Department of Parasitology, Faculty of Medicine, Kanazawa University, Kanazawa City 920, Japan
Taka-Aki Ohyama
Affiliation:
Department of Parasitology, Faculty of Medicine, Kanazawa University, Kanazawa City 920, Japan
Kaoru Kondo
Affiliation:
Department of Parasitology, Faculty of Medicine, Kanazawa University, Kanazawa City 920, Japan
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Abstract

To increase our understanding of the immune response to Anisakis infection, antigen specific IgG, IgA and IgE responses were identified using an immunoblot technique after polyacrylamide gel electrophoresis of excretory-secretory products from the larval stage of Anisakis simplex. Nine sera were drawn from proven cases of gastric anisakiasis within 3 days after symptoms had developed. The molecular weight of the major antigenic bands were distributed between 50 kDa and 120 kDa of the antigens. In nine cases of gastric anisakiasis, three of them were positive for IgG response, five for IgE, and six for IgA, respectively. None of control sera recognized the antigenic bands in IgA and IgE responses. In contrast, two controls had IgG antibodies against 1–2 proteins in the 65–95 kDa region. The antigenicity of the excretory–secretory products was lost following treatment by 0·2% trypsin, but not by 0·2 M periodic acid. Based on the results of reactivity to lectins, antigenic bands of the ES products possessed mucin type glycoconjugate residues in their protein portion. This indicates that the humoral responses of IgA and IgE antibodies to the larval ES antigens are a more reliable index of infection than that of the IgG response.

Keywords

Anisakis simplexES antigensIgAIgEimmune responseimmunoblotting
Type
Research Article
Information
Journal of Helminthology , Volume 64 , Issue 4 , December 1990 , pp. 310 - 318
Copyright
Copyright © Cambridge University Press 1990

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References

REFERENCES

Akao, N. & Kondo, K. (1986) Glycoconjugates of excretory-secretory antigens of second stage larvae of Toxocara canis: Analysis of their reactivity to lectins. Japanese Journal of Parasitology, 35, 395–401.Google Scholar
Akao, N. & Yoshimura, H. (1989) Latex agglutination test for immunodiagnosis of gastric anisakiasis. In: Gastric anisakiasis in Japan, epidemiology, diagnosis, treatment (editors, Ishikura, H. and Namiki, M.), pp. 97–102. Springer-Verlag Ltd.: Tokyo.CrossRefGoogle Scholar
Desowitz, R. S., Raybourne, R. B., Ishikura, H. & Kliks, M. M. (1985) The radioallergosorbent test (RAST) for the serological diagnosis of human anisakiasis. Transactions of the Royal Society of Tropical Medicine & Hygiene, 79, 256–259.CrossRefGoogle ScholarPubMed
Hancock, K. & Tsang, V. C. W. (1983) India ink staining of proteins on nitrocellulose paper. Analytical Biochemistry, 133, 157–162.CrossRefGoogle ScholarPubMed
Kagei, N. & Kureha, N. (1970) List of the larvae of Anisakis spp. recorded from marine fish and squids caught off the Japan and its offshore islands. Bulletin of the Institute of Public Health, 19, 76–85.Google Scholar
Kawauchi, H., Namiki, M., Morooka, T., Nakagawa, K. & Oguro, T. (1973) Gastric anisakiasis presenting acute gastrointestinal symptoms—with special reference to the endoscopic and roentgenographic findings of Anisakis larva penetrating into the wall of the human stomach and to its clinical features. Stomach and Intestine, 8, 31–38.Google Scholar
Kijimoto, S., Katagiri, Y. & Ochiai, H. (1985) Analysis of N-linked oligosaccharide chains of glycoproteins on nitro-cellulose sheets using lectin-peroxidase reagents. Analytical Biochemistry, 147, 222–229.CrossRefGoogle Scholar
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.CrossRefGoogle ScholarPubMed
Lowry, O., Rosenbough, N. J., Farr, A. L. & Randall, R. J. (1951) Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193, 265–275.CrossRefGoogle ScholarPubMed
Namiki, M., Morooka, T., Kawauchi, H., Ueda, N., Sekiya, C, Nakagawa, K., Furuta, T., Oguro, T. & Kamata, H. (1970) Diagnosis of acute gastric anisakiasis. Stomach and Intestine, 5, 1437–1440.Google Scholar
Namiki, M. & Yazaki, Y. (1989) Treatment of gastric anisakiasis with acute symptoms. In: Gastric anisakiasis in Japan, epidemiology, diagnosis, treatment (editors, Ishikura, H. and Namiki, M.), pp. 129–131. Springer-Verlag Ltd.: Tokyo.CrossRefGoogle Scholar
Ohtaki, H. & Ohtaki, R. (1989) Clinical manifestation of gastric anisakiasis. In: Gastricanisakiasis in Japan, epidemiology, diagnosis, treatment (editors, Ishikura, H. and Namiki, M.), pp. 37–46. Springer-Verlag Ltd.: Tokyo.Google Scholar
Raybourne, R., Deardorff, T. L. & Bier, J. W. (1986) Anisakis simplex: Larval excretory secretory protein production and cytostatic action in mammalian cell cultures. Experimental Parasitology, 62, 92–97.CrossRefGoogle ScholarPubMed
Suzuki, T., Shiraki, T. & Otsuru, M. (1969) Studies on the immunological diagnosis of anisakiasis: II. Isolation and purification of anisakis antigen. Japanese Journal of Parasitology, 18, 232–240.Google Scholar
Takahashi, S., Sato, S. & Ishikura, H. (1986) Establishment of monoclonal antibodies that discriminate the antigen distribution specifically found in Anisakis larvae (type 1). Journal of Parasitology, 72, 960–962.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 of Sciences of the United States of America, 76, 4350–4354.CrossRefGoogle ScholarPubMed
Yagihashi, A., Sato, N., Takahashi, S., Ishikura, H., Kondo, A., Okubo, M., Torimoto, K., Hayasaka, H. & Kikuchi, K. (1989) Establishment of a sero-diagnostic assay system for anisakiasis using a monoclonal antibody specific for Anisakis larval antigen. Journal of Clinical & Experimental Medicine, 148, 213–214.Google Scholar
Yoshimura, H., Akao, N., Kondo, K. & Ohnishi, Y. (1979) Clinicopathological studies on larval anisakiasis, with special reference to the report of extra-gastrointestinal anisakiasis. Japanese Journal of Parasitology, 28, 347–354.Google Scholar
Yoshimura, H., Akao, N., Kondo, K., Onhishi, Y., Funaoka, H. & Yamane, K. (1980) Two cases of extra-gastrointestinal anisakiasis and evaluation of immunodiagnosis. Japanese Journal of Clinical Pathology, 28, 708–712.Google ScholarPubMed
Yoshimura, H., Kondo, K., Ohnishi, Y. & Akao, N. (1983) Studies on immunodiagnosis of anisakiasis. Report for Scientific Research Grant from Ministry of Education of Japan.Google Scholar