Identification of excretory-secretory products of larval and adult Ostertagia ostertagi by immunoscreening of cDNA libraries.
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.
Vercauteren
Isabel
I
Department of Parasitology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, Merelbeke B-9820, Belgium. isabel.vercauteren@rug.ac.be
Geldhof
Peter
P
Peelaers
Iris
I
Claerebout
Edwin
E
Berx
Geert
G
Vercruysse
Jozef
J
D000818:Animals; D000909:Antibodies, Helminth; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling; D015723:Gene Library; D017238:Genes, Helminth; D015801:Helminth Proteins / Q000032:analysis / Q000235:genetics*; D008969:Molecular Sequence Data; D010028:Ostertagia / Q000235:genetics / Q000502:physiology*; D014819:Vitellogenins / Q000235:genetics
D000909:Antibodies, Helminth; D018076:DNA, Complementary; D015801:Helminth Proteins; D014819:Vitellogenins