Variation of antigenic proteins of eggs and developmental stages of Paragonimus westermani
Y Kong,1J Y Chung,1D H Yun,1L S Kim,2S Y Kang,2A Ito,3L Ma,3 and S Y Cho*4
1Department of Parasitology, Sungkyunkwan University, School of Medicine, Suwon, Korea.
Received June 13, 1997; Accepted August 14, 1997.
Diagnosis of early paragonimiasis is difficult because parasitological evidence is not easily obtained. Antibody tests have been proposed as a good substitute for classical diagnostic techniques. Using the crude extracts of Paragonimus westermani eggs, metacercariae, 4- and 7-week juveniles, and 16-week adults as antigens, we observed the early antibody responses. Sera were obtained from 4 experimental cats, fed 50 metacercariae each, at intervals until 13 weeks post-infection. Antibody (IgG) responses were identified by ELISA using extracts of 4-week juveniles, followed by those of 7- and 16-week worms. Antibody responses were minimal against the metacercarial extracts. Antibodies to P. westermani egg extracts were elevated after 10 weeks post-infection. In immunoblot analysis, more than nine protein bands in 4-week juveniles reacted with the early infection sera. Antigenic proteins in adult worms were different from those of juveniles. After four weeks of infection, 32 and 35 kDa bands in the adult extracts were increasingly reactive. Egg specific proteins at 28, 46 and 94 kDa were reactive only after 10 weeks. Antigenic components reacting to the early infection sera changed during the maturation stages of P. westermani; almost all juvenile antigens were replaced by adult antigen components.
Fig. 1 Antibody levels in the sera of early experimental cat paragonimiasis (mean-S.D., n=4) measured against five different antigenic preparations of P. westermani.
Fig. 2 Analysis of the crude extracts of different stages of P. westermani as seen by SDS-PAGE (Coomassie blue stained). Mr, molecular weight in kDa; MC*, metacercariae.
Fig. 3 Immunoblot analysis of the crude extracts of eggs (lanes Egg), 4-week old juveniles (lanes 4-wk) and 16-week old adults (lanes 16-wk) using the cat sera collected until 13-week of infection. Top indicates infection age of the examined sera. Mr, molecular weight in kDa. The experimental cat sera (n=4) were pooled, diluted at 1:400 in Tris-HCI containing 2% casein (w/v) and probed overnight. A 1:1,000 diluted peroxidase conjugated anti-cat IgG was used as a secondary antibody.
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