Immunological studies on an Onchocerca volvulus intermediate-filament protein.
We report the complete sequence of the cDNA encoding an intermediate filament (IF) protein from Onchocerca volvulus. The OVIF cDNA encodes a protein of 613 amino acid residues, which has a predicted molecular weight of approximately 70 kD. The size of the protein encoded by the OVIF cDNA corresponds well with estimates obtained in Western blotting experiments, but these same experiments suggest that Onchocerca sp. may contain at least two IF proteins. To identify the domain(s) of the OVIF protein responsible for immunogenicity in human filariasis patients, fragments of the OVIF cDNA were subcloned into the appropriate pGEX vector and Western blots of the corresponding fusion proteins probed with human sera. Pooled sera from general onchocerciasis or Bancroftian filariasis patients reacted only with the C-terminal region of the OVIF protein, whereas sowda onchocerciasis sera reacted 1-2 orders of magnitude more strongly and with a number of other epitopes. Immunolocalization experiments implied that the OVIF protein is associated with both muscle and a number of specific membrane layers. These studies provide the basis for evaluation of the protective potential of the OVIF protein.
Zhang
D
D
Department of Molecular Sciences, James Cook University of North Queensland, Townsville, Australia.
Miller
D J
DJ
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology; D000947:Antigens, Helminth / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D003429:Cross Reactions / Q000276:immunology; D018076:DNA, Complementary / Q000032:analysis; D005368:Filariasis / Q000276:immunology; D005455:Fluorescent Antibody Technique; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D017181:Onchocerca volvulus / Q000235:genetics / Q000276:immunology*; D009855:Onchocerciasis / Q000276:immunology; D014958:Wuchereria bancrofti / Q000276:immunology
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018076:DNA, Complementary; D015801:Helminth Proteins; D016056:Immunodominant Epitopes; D007381:Intermediate Filament Proteins