Characterization of a muscle-associated antigen from Wuchereria bancrofti.
A recombinant clone, WbN1, isolated from a genomic expression library of Wuchereria bancrofti and showing restricted specificity at the DNA level (Southern and PCR analyses) for Wuchereria bancrofti and Brugia malayi has been previously described. Sequence analysis of WbN1 indicated that it had notable similarity to myosin. Further characterization using in situ hybridization has localized the mRNA in the muscle of the adult parasite and in the microfilariae. Rabbit polyclonal antiserum, raised against the recombinant WbN1 fused to the maltose-binding protein, recognized a 200-kDa polypeptide in immunoblots containing B. malayi antigen extracts. The same antibody also recognized myosin extracted from Brugia pahangi, Onchocerca volvulus, and Caenorhabditis elegans. Localization using the rabbit antiserum revealed the presence of the antigen in the adult muscle tissue and in the microfilariae; the same antibody inhibited the binding of a monoclonal antibody 28.2 (directed toward MHC B of C. elegans myosin) to the recombinant WbN1 antigen and also to purified C. elegans myosin. Based on homology data, structural location, competitive ELISA, and immunoblot we conclude that WbN1 is related to myosin or a similar myofibrillar protein.
Raghavan
N
N
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Maina
C V
CV
Fitzgerald
P C
PC
Tuan
R S
RS
Slatko
B E
BE
Ottesen
E A
EA
Nutman
T B
TB
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry*; D004605:Elephantiasis, Filarial / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005260:Female; D005368:Filariasis / Q000276:immunology; D015801:Helminth Proteins / Q000032:analysis / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009218:Myosins / Q000032:analysis / Q000737:chemistry* / Q000276:immunology; D011657:Pulmonary Eosinophilia / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D014958:Wuchereria bancrofti / Q000276:immunology*
D000947:Antigens, Helminth; D000939:Epitopes; D015801:Helminth Proteins; D011993:Recombinant Fusion Proteins; C079595:myosin-like protein, Wuchereria bancrofti; D009218:Myosins