Recombinant dissection of myosin heavy chain of Toxocara canis shows strong clustering of antigenic regions.
Myosins from nematode parasites elicit strong humoral and cellular immune responses and have been investigated as vaccine candidates. In this study we cloned and sequenced a cDNA coding for myosin heavy chain from Toxocara canis, a nematode parasite of canids which may also infect humans and cause various unspecific symptoms. To determine the major antigenic regions the myosin heavy chain was systematically dissected into ten overlapping recombinant fusion polypeptides which were purified by metal chelate chromatography. Single fragments were then tested for their IgG reactivity in sera from toxocarosis patients and healthy probands. Two regions, one region at the mid to carboxy-terminal end of the head domain and one region in the rod domain, were identified as major antigens, which in combination were positive with 86% of the sera. The other domains were less reactive. This shows that the patients' IgG reactivity was not directed evenly against all parts of the molecule, but was rather clustered in few regions.
Obwaller
A
A
Department of Pathophysiology, University of Vienna, Austria.
Duchêne
M
M
Bruhn
H
H
Steipe
B
B
Tripp
C
C
Kraft
D
D
Wiedermann
G
G
Auer
H
H
Aspöck
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000032:analysis; D018509:DNA, Helminth / Q000032:analysis; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D018995:Myosin Heavy Chains / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D017422:Sequence Analysis, DNA; D017166:Toxocara canis / Q000276:immunology*; D014120:Toxocariasis / Q000097:blood
D000947:Antigens, Helminth; D017931:DNA Primers; D018076:DNA, Complementary; D018509:DNA, Helminth; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D018995:Myosin Heavy Chains