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Characterization of a major surface-associated excretory–secretory antigen of Trichinella spiralis larvae with antibodies to keyhole limpet haemocyanin

Published online by Cambridge University Press:  06 April 2009

J. Modha ,
W. M. Robertson ,
M. W. Kennedy  and
J. R. Kusel
J. Modha
Affiliation:
Department of Biochemistry, University of Glasgow, Glasgow G12 8QQ
W. M. Robertson
Affiliation:
Department of Zoology, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA
M. W. Kennedy
Affiliation:
Wellcome Laboratories for Experimental Parasitology, University of Glasgow, Bearsden, Glasgow G61 1QH
J. R. Kusel
Affiliation:
Department of Biochemistry, University of Glasgow, Glasgow G12 8QQ
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Summary

A multi-subunit antigen (native Mr > 200 kDa, reduced Mr 97–100 kDa) has been identified in homogenates of Trichinella spiralis larvae using affinity-purified rabbit anti-keyhole limpet haemocyanin (KLH) antibodies and its cross-reactivity with KLH was confirmed by competition blotting. The antigen was not present at the larval surface but was exposed after treatment of the larvae with the detergent cetyltrimethyl ammonium bromide (CTAB) which removed the surface coat. This correlated with a significant decrease in insertion of the surface-restricted fluorescent lipid probe AF18, indicating that the surface coat must be lipidic in nature. Unlike KLH, the larval antigen blotted onto nitrocellulose was itself periodate insensitive. Periodate treatment of whole larvae, however, resulted in shedding of the surface, to which anti-KLH antibodies then bound intensely. Anti-KLH antibodies also recognized three (49, 55, 108 kDa) of the four most dominant antigens in excretory–secretory (ES) products of cultured larvae, whose excretion–secretion was increased with CTAB. The nature, location and function of the antigen is discussed.

Keywords

Trichinella spiraliskeyhole limpet haemocyaninfluorescent lipid probeoctadecyl aminofluorescein (AF18)excretory–secretory antigens
Type
Research Article
Information
Parasitology , Volume 109 , Issue 4 , November 1994 , pp. 531 - 538
Copyright
Copyright © Cambridge University Press 1994

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