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J. Biol. Chem., Vol. 278, Issue 19, 16941-16951, May 9, 2003
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From the Haemonchus contortus is a
nematode that infects small ruminants. It releases a variety of
molecules, designated excretory/secretory products (ESP), into the
host. Although the composition of ESP is largely unknown, it is a
source of potential vaccine components because ESP are able to induce
up to 90% protection in sheep. We used proteomic tools to analyze ESP
proteins and determined the recognition of these individual proteins by
hyperimmune sera. Following two-dimensional electrophoresis of ESP,
matrix-assisted laser desorption ionization time-of-flight and
liquid chromatography-tandem mass spectrometry were used for protein
identification. Few sequences of H. contortus have
been determined. Therefore, the data base of expressed sequence tags
(dbEST) and a data base consisting of contigs from
Haemonchus ESTs were also consulted for identification. Approximately 200 individual spots were observed in the two-dimensional gel. Comprehensive proteomics analysis, combined with bioinformatic search tools, identified 107 proteins in 102 spots. The data include known as well as novel proteins such as serine, metallo- and aspartyl proteases, in addition to H. contortus ESP components like
Hc24, Hc40, Hc15, and apical gut GA1 proteins. Novel proteins were
identified from matches with H. contortus ESTs displaying
high similarity with proteins like cyclophilins, nucleoside diphosphate
kinase, OV39 antigen, and undescribed homologues of
Caenorhabditis elegans. Of special note is the finding of
microsomal peptidase H11, a vaccine candidate previously regarded as a
"hidden antigen" because it was not found in ESP. Extensive
sequence variation is present in the abundant Hc15 proteins. The Hc15
isoforms are differentially recognized by hyperimmune sera, pointing to
a possible specific role of Hc15 in the infectious process and/or in
immune evasion. This concept and the identification of multiple
novel immune-recognized components in ESP should assist future
vaccine development strategies.
Comprehensive Analysis of the Secreted Proteins of the
Parasite Haemonchus contortus Reveals Extensive Sequence
Variation and Differential Immune Recognition*
§¶,
**,¶,**, and¶
Division of Parasitology and Tropical
Veterinary Medicine, Department of Infectious Diseases and Immunology,
Utrecht University, P.O. Box 80165, 3508 TD, Utrecht, The Netherlands
and the Department of Biomolecular Mass Spectrometry, Bijvoet
Center for Biomolecular Research and Utrecht Institute for
Pharmaceutical Sciences, Utrecht University, Sorbonnelaan 16, 3584 CA, Utrecht, The Netherlands
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.:
31-30-253-2582; Fax: 31-30-254-0784; E-mail: e.vries@vet.uu.nl.
Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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