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Immune reactions between excretory-secretory antigens and specific antibodies of Clonorchis sinensis before and after praziquantel treatment in experimentally infected rabbits
S I Kim*
Department of Parasitology, Chosun University Medical College, Kwangju 501-759, Korea.
Received January 03, 1994; Accepted February 28, 1994.
Abstract
This study was designed to evaluate the humoral immune reactions in clonorchiasis before and after praziquantel treatment. Rabbits were infected with 150 or 450 metacercariae, treated on 4 and 8½ months after infection, and observed for 13 months of posttreatment. Infection controls were maintained for 22 months. Antigen was the metabolic product of worms incubated in physiologic saline. The immune reactions of anti- Clonorchis IgG were observed using SDS-PAGE/immunoblot. During the infection and posttreatment, the antigenic proteins of 66, 63, 54, 52, 50, 47, 42, 40, 38, 34, 33, 30, 27, 25, 23, 20, 19, 18, 17, 16, 15, 14, 13, 12.5 and 11.5 kDa were detected. Of them, 33, 27, 13, and 12.5-kDa antigens were highly antigenic and observed predominantly in infection controls. After the treatment, 13 and 12.5-kDa antigens faded in 6 months after the second treatment, but 33 and 27-kDa antigens were detected until 13 months of posttreatment. The results clearly demonstrate that 13 and 12.5-kDa antigens represent attenuated host immune reactions by praziquantel treatment. As the 12.5-kDa antigen had a large amount of protein in SDS-PAGE, it was designated as 'K2-Ag' of C. sinensis.
Figures
Fig. 1 Electrophoretic pattern of component proteins of excretory-secretory antigen of Clonorchis sinensis (CsE). SDS-PAGE was conducted in 10-15% linear gradient gel (GEL). Each band was detected by silver staining. Proteins of CsE were electrophoretically transferred from gel to nitrocellulose (NC), and stained by 0.1% amido black 10B. Mr means standard marker proteins, numerals represent molecular masses in kDa.
Fig. 2 SDS-PAGE/immunoblot patterns in immune reactions of anti-Clonorchis rabbit IgG antibodies to excretory-secretory antigens of C. sinensis during infection and posttreatment. I means experimental infection of 450 metacercariae, t & T were the indication of the first & the second medication of praziquantel. As the first medication failed, the second treatment was needed. Beneath these markings, numerals represent months after infection, the first, and the second treatment. Mr indicates standard markers.
Fig. 3 Immune reactions between excretory-secretory antigens and specific IgG antibodies of C. sinensis before and after praziquantel treatment in experimentally infected rabbits. (1) 450 metacercariae-infected and treated, (2) 150 metacercariae-infected and treated, and (3) 450 or 150 metacercariae-infected and untreated rabbits were compared in their immune reactions according to SDS-PAGE/immunoblot. In the treated rabbits, 26 antigens were detected and grouped into A, B, C, D, E, and K. In untreated group, the most predominent antigens were 33, 27, and 12.5-kDa antigens, After the praziquantel treatment, 12,5-kDa antigen was faintly detected on the 6th month after the second treatment, while 33 and 37-kDa antigens were observed until 13 months after the second treatment.
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