Warning: mkdir(): Permission denied in /home/virtual/lib/view_data.php on line 81

Warning: fopen(upload/ip_log/ip_log_2024-03.txt): failed to open stream: No such file or directory in /home/virtual/lib/view_data.php on line 83

Warning: fwrite() expects parameter 1 to be resource, boolean given in /home/virtual/lib/view_data.php on line 84
Tissue origin of soluble component proteins in saline extract of adult Paragonimus westermani
| Home | E-Submission | Sitemap | Contact us |  
top_img
Korean J Parasito Search

CLOSE

Korean J Parasito > Volume 30(2):1992 > Article

Original Article
Korean J Parasitol. 1992 Jun;30(2):91-100. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1992.30.2.91
Copyright © 1992 by The Korean Society for Parasitology
Tissue origin of soluble component proteins in saline extract of adult Paragonimus westermani
Y Kong,C Y Park,**S Y Kang and S Y Cho
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.

***Junior medical student of Chung-Ang University at the time of the study

Abstract

Tissue origin of individual component proteins in crude extract of adult Paragonimus westermani was investigated. Major soluble component proteins were separated by disc-PAGE in 8% slab gel. By predefined Rf values, strips of gel containing each band protein was cut out. Each band protein was eluted by electrophoresis. Monospecific antibodies were prepared by immunizing rabbits with each band protein. When peroxidase-antiperoxidase (PAP) staining was done, antiserum to Band 1 reacted to content of eggs both in the worm and in the infected lung tissue. Antiserum to Band 2 reacted to parenchymal tissue of the worm. Antiserum to Band 4 showed the positive reaction at intestinal content while that to Band 5 reacted to the intestinal epithelial border. Antiserum to combined proteins of Bands 6/7 and that to Band 8 reacted to parenchymal tissue of the worm respectively. From the results, the origin of individual proteins in crude extract of adult P. westermani could be differentiated.

Figures


Fig. 1
Purity of the band proteins of adult P. westermani after the disc-PAGE and electrophoretic elution. The proteins were visualized in 8% disc-PAGE. Coomassie stained. Numericals stand for an each band protein.

Mr : Molecular mass in kDa, C: Crude extract.



Fig. 2
SDS-PAGE fingings of the band protein. A gradient separationg gel of 10~15% sas used. Markings and numericals are the same as described in Fig. 1.


Figs. 3-8
Figs. 3 & 4. Negative control I & II (×100). The worm sections were treated with PBS (I) and rabbit serum collected before immunization (II) as primary antibodies. No structure showed the positive reactions.

Fig. 5. Positive control (×100). Antiserum against the crude extract was reacted at intestinal epithelial border, intestinal content, eggs and parenchymal tissue. Cvary and vitellarian follicles showed equivocal reaction. Tegument, testes, basal layer of intestine and suckers were not stained positively with PAP.

Fig. 6. The rabbit antiserum to Band 1 reacted to the cells within egg shells in uterus (×100).

Fig. 7. Another PAP staining of antiserum to Band 1 (×100). The eggs which were scattered in cat lung tissue showed the stronger positive reaction.

Fig. 8. When the antiserum to Band 2 was reacted, positive reaction was observed at the parenchymal tissue of the worm (×100).



Figs. 9-12
Fig. 9. Intestinal content was stained positively with antiserum to Band 4(×100).

Fig. 10. Antiserum to Band 5 reacted to the intestinal epithelial border. Basal part of epithelium was not stained with chromogen (×100).

Fig. 11. Rabbit antiserum to Bands 6/7 showed the diffuse positive reaction to parenchymal tissue of the worm (×200).

Fig. 12. Parenchymal tissur of the worm was stained with antiserum to Band 8 (×100).


Tables


Table 1
Recovery of each Band protein when 45 mg of protein in the crude extract of adult P. westermani was purified by disc-PAGE and electrophoretic elution


Table 2
Specific IgG antibody levels (abs. by ELISA) in differently diluted rabbit antisera against respective antigens

References
1. Cho SY, Hong ST, Rho YH, Choi SY, Han YC. Application of micro-ELISA in serodiagnosis of Human paragonimiasis. Korean J Parasitol 1981;19(2):151–156.
 
2. Cho SY, Kim SI, Kang SY, Kong Y, Han SK, Shim YS, Han YC. Antibody changes in paragonimiasis patients after praziquantel treatment as observed by ELISA and immunoblot. Korean J Parasitol 1989;27(1):15–21.
 
3. Choi WY, Yoo JE, Nam HW, Choi HR. [Purification of antigenic proteins of Paragonimus westermani and their applicability to experimental cat paragonimiasis]. Korean J Parasitol 1986;24(2):177–186.
 
4. Damonneville M, Pierce RJ, Verwaerde C, Capron A. Allergens of Schistosoma mansoni. II. Fractionation and characterization of S. mansoni egg allergens. Int Arch Allergy Appl Immunol 1984;73(3):248–255.
  
5. Deelder AM, De Jonge N, Boerman OC, Fillie YE, Hilberath GW, Rotmans JP, Gerritse MJ, Schut DW. Sensitive determination of circulating anodic antigen in Schistosoma mansoni infected individuals by an enzyme-linked immunosorbent assay using monoclonal antibodies. Am J Trop Med Hyg 1989;40(3):268–272.
 
6. Imai J, et al. Jpn J Parasitol 1988;37:398–404.
7. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227(5259):680–685.
  
8. Joo KH, Hong SC, Chung MS, Rim HJ. [Analysis of antigenic specificities of Paragonimus westermani developmental stages using immunoblot technique]. Korean J Parasitol 1989;27(1):1–7.
 
9. Joo KH, Ahn H, Chung MS, Rim HJ. Demonstration of species-specific and cross reactive components of Paragonimus westermani crude worm antigen by EITB. Korean J Parasitol 1989;27(1):9–14.
 
10. Kang SY, Kong Y, Cho SY. Component proteins in crude extract of adult Paragonimus westermani purified by immunoaffinity chromatography using monoclonal antibodies. Korean J Parasitol 1991;29(4):363–369.
 
11. Kim DC, et al. Rep NIH Korea 1983;19:109–114.
12. Kim SH, Kong Y, Kim SI, Kang SY, Cho SY. Immunoblot observation of antigenic protein fractions in Paragonimus westermani reacting with human patients sera. Korean J Parasitol 1988;26(4):239–243.
 
13. Kim SI, Ko EK, Kang SY, Cho SY. [Antigenicity of the soluble egg antigen of Paragonimus westermani]. Korean J Parasitol 1986;24(1):49–54.
 
14. Kong Y, Kim WB, Kang SY, Cho SY. Molecular weight of major component proteins in crude saline extract of adult Paragonimus westermani. Korean J Parasitol 1991;29(2):113–120.
 
15. Kwon OS, Lee JS, Rim HJ, Kim SJ. [Antigenic localities in the tissues of the young adult worm of Paragonimus westermani using immunogold labeling method]. Korean J Parasitol 1991;29(1):31–41.
 
16. Lee OR, Chang JK. [ELISA of paragonimiasis in cat by crude and purified antigens of Paragonimus westermani]. Korean J Parasitol 1986;24(2):187–193.
 
17. Lee SH, Sung SH, Chai JY. [Immunohistochemical study on the antigenicity of body compartments of Paragonimus westermani]. Korean J Parasitol 1989;27(2):109–117.
 
18. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(1):265–275.
 
19. McKerrow JH, Doenhoff MJ. Schistosome proteases. Parasitol Today 1988;4(12):334–340.
  
20. Shim YS, et al. Seminars in Resp Med 1991;12:35–45.
 
21. Slemenda SB, Maddison SE, Jong EC, Moore DD. Diagnosis of paragonimiasis by immunoblot. Am J Trop Med Hyg 1988;39(5):469–471.
 
22. Sugiyama H, Hinoue H, Katahira J, Horiuchi T, Tomimura T, Kamata Y, Kozaki S. Production of monoclonal antibody to characterize the antigen of Paragonimus westermani. Parasitol Res 1988;75(2):144–147.
  
23. Sugiyama H, Sugimoto M, Akasaka K, Horiuchi T, Tomimura T, Kozaki S. Characterization and localization of Paragonimus westermani antigen stimulating antibody formation in both the infected cat and rat. J Parasitol 1987;73(2):363–367.
  
24. Waterborg JH, et al. Method in Molec Biol 1984;1:157–164.
25. Yokogawa M, et al. Jpn J Parasitol 1983;32:317–322.
Editorial Office
Department of Molecular Parasitology, Samsung Medical Center, School of Medicine, Sungkyunkwan University,
2066 Seobu-ro, Jangan-gu, Suwon 16419, Gyeonggi-do, Korea.
Tel: +82-31-299-6251   FAX: +82-1-299-6269   E-mail: kjp.editor@gmail.com
About |  Browse Articles |  Current Issue |  For Authors and Reviewers
Copyright © 2024 by The Korean Society for Parasitology and Tropical Medicine.     Developed in M2PI