Identification of excretory-secretory products of larval and adult Ostertagia ostertagi by immunoscreening of cDNA libraries

被引:49
作者
Vercauteren, I
Geldhof, P
Peelaers, I
Claerebout, E
Berx, G
Vercruysse, J
机构
[1] State Univ Ghent, Dept Parasitol, Fac Med Vet, B-9820 Merelbeke, Belgium
[2] State Univ Ghent, Dept Mol Biomed Res, Fac Sci, B-9000 Ghent, Belgium
[3] VIB, B-9000 Ghent, Belgium
关键词
Ostertagia ostertagi; immunoscreening; excretory-secretory products; candidate protective antigens;
D O I
10.1016/S0166-6851(02)00274-8
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Excretory-secretory (ES) products of Ostertagia ostertagi, an abomasal nematode of cattle, are considered to be important for the development and survival of the parasite within the host. To gain insight in the composition of these ES products of both larval (L3, L4) and adult life stages of Ostertagia cDNA libraries of the parasite were immunoscreened with polyclonal rabbit serum raised against these ES products. This approach led to the identification of 41 proteins, amongst which are structural proteins such as actin, kinesin and vitellogenin, housekeeping proteins such as those involved in protein folding, different metabolic pathways or mitochondrial functioning and proteins associated with stress (heat shock protein) or antioxidantia (thioredoxin peroxidase). A large number of the isolated proteins were similar to hypothetical proteins of the model nematode Caenorhabditis elegans. Because somatic proteins can be non-specifically released during in vitro culturing as nematodes deteriorate, it was checked if the isolated proteins are genuinely secreted. The amino acid sequences of the translated cDNAs were investigated for signal peptides and monospecific antibodies against the isolated proteins were purified and used to develop Western blots of ES and somatic extracts. In this manner it could be proven that 15 cDNAs code for genuine secreted proteins. The identification of these ES antigens allows to select proteins with potential protective capacities, which are targets for vaccine development. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:201 / 208
页数:8
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