Identification, characterization and expression of Toxocara canis nematode polyprotein allergen TBA-1

We have cloned the cDNA of TBA-1, the Nematode polyprotein allergen (NPA) of Toxocara canis and found it to be most similar to ABA-1, the Ascaris NPA, on the basis of amino acid sequence. We could study the antigenic properties of an E-coli synthesized fusion protein prepared with the cloned gene since no glycosylation site was expected from the deduced amino acid sequence. Although no IgE responses to TBA-1 were detected, recombinant TBA-1 was differently recognized by serum IgG antibodies when the recombinant TBA-1 was directly adsorbed vs when immobilized via a streptavidin linkage on polystyrene microtitre wells. One group of sera recognized TBA-1 directly immobilized while the second only recognized TBA-1 immobilized via streptavidin linkage. The former were from rodents immunized with a Toxocara sp. adult worm extract while the latter were obtained from rodents infected with T. canis larva or immunized with a Anisakis simplex L3 larval extract. These observations suggest that the two in vivo forms of TBA-1 are expressed, but during different stages of the parasite's life cycle.