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Korean J Parasitol > Volume 17(1):1979 > Article

Original Article
Korean J Parasitol. 1979 Jun;17(1):73-80. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1979.17.1.73
Copyright © 1979 by The Korean Society for Parasitology
Immunoelectrophoresis for Fasciola hepatica
Won-Young Choi and Ok-Ran Lee
Department of Parasitology, Catholic Medical College and Catholic Institute For Parasitic Diseases, Korea.
Abstract

In an attempt to investigate the specific antigenic substance of Fasciola hepatica, Ouchterlony tests and immunoelectrophoretic analyses were carried out. Crude Fasciola antigen was prepared and fractionated by Sephadex G-200 column to Antigen I, II and III according to protein content. Crude antigens of Paragonimus westermani, Clonorchis sinensis and Paramphistomum sp. were also prepared for control and absorption study. Antiserum was prepared by injecting 0.5 ml of crude Fasciola antigen with same amount of complete Freund's adjuvant in rabbits, 10 times at an interval of l week.

The results obtained in this study were as follows:

1. Crude Fasciola antigen reacted with antiserum with 9 precipitin bands by Ouchterlony test and with 11 bands by immunoelectrophoresis.

2. Cross reaction was observed between Paragonimus, Clonorchis and Paramphistomum antigens and anti-Fasciola rabbit serum respectively. By Ouchterlony test, 3-4 cross reacting bands were found.

3. Anti-Fasciola sera which were absorbed with respective Paragonimus, Clonorchis and Paramphistomum antigens, reacted with Fasciola crude antigen. Ouchterlony test gave 5-6 precipitin bands. Further reaction between Fasciola antigen and antiserum absorbed with the above 3 antigens concomitantly gave 5 precipitin bands by Ouchterlony test and 7 bands by immunoelectrophoretic analyses.

4. Fractionated Fasciola antigens (Antigens I, II and III) reacted with anti-Fasciola rabbit serum in immunoelectrophoresis. Antigen I, II and III gave 2, 3 and 5 precipitin bands respectively. Anti-Fasciola rabbit serum which was absorbed with 3 trematodes antigens gave, by immunoelectrophoresis, 1 band with Antigen I, 2 bands with Antigen II and III of Fasciola hepatica.

From the above results, it is concluded that Fasciola hepatica possessed the specific antigenic substance not cross-reacted with other trematodes.

Figures


Fig. 1
Fractionation of Fasciola hepatica crude antigen on Sephadex G-200.


Fig. 2
Central well: F. hepatica crude antigen

Outer wells Control: normal rabbit serum

1,2,3,4 & 5: Sensitized anti-F. hepatica rabbit sera.



Fig. 3
Central well: F. hepatica crude antigen

Trough: Sensitized anti-F. hepatica rabbit sera



Fig. 4
Central well: Sensitized anti-F. hepatica rabbit serum

1 & 4: C. sinensis antigen

2 & 5: Paramphistomum antigen

3 & 6: P. westermani antigen



Figs. 5-7
Fig. 5. Central well: anti-F. hepatica rabbit absorbed with P. westermani antigen, Outer well: F. hepatica crude antigen

Fig. 6. Central well: anti-F. hepatica serum absorbed with P. westermani antigen, Outer well: F. hepatica crude antigen

Fig. 7. Central well: anti-F. hepatica serum absorbed with Paramphistomum antigen, Outer well: F. hepatica crude antigen



Fig. 8
Central well: anti-F. hepatica serum absorbed with P. westermani, C. sinensis and Paramphistomum

Outer wells: F. hepatica crude antigen



Fig. 9
Well: F. hepatica crude antigen

Trough: anti-F. hepatica serum absorbed with P. westermani, C. sinensis and Paramphistomum

Outer wells: F. hepatica crude antigen



Figs. 10-12
Fig. 10. Central well: anti-F. hepatica serum, Outer wells: 1st fraction of F. hepatica antigen

Fig. 11. Central well: anti-F. hepatica serum, Outer wells: 2nd fraction of F. hepatica antigen

Fig. 12. Central well: anti-F. hepatica serum, Outer wells: 3rd fraction of F. hepatica antigen



Figs. 13-15
Fig. 13. Rt well: 1st fraction of F. hepatica antigen, Lt well: F. hepatica crude antigen, Trough: anti-F. hepatica serum

Fig. 14. Rt well: 2nd fraction of F. hepatica antigen, Lt well: F. hepatica crude antigen, Trough: anti-F. hepatica serum

Fig. 15. Rt well: 3rd fraction of F. hepatica antigen, Lt well: F. hepatica crude antigen, Trough: anti-F. hepatica serum



Figs. 16-18
Fig. 16. Rt well: 1st fraction of F. hepatica antigen, Lt well: F. hepatica crude antigen, Trough: absorbed anti-F. hepatica serum

Fig. 17. Rt well: 2nd fraction of F. hepatica antigen, Lt well: F. hepatica crude antigen, Trough: absorbed anti-F. hepatica serum

Fig. 18. Rt well: 3rd fraction of F. hepatica antigen, Lt well: F. hepatica crude antigen, Trough: absorbed anti-F. hepatica serum


References
1. Choi WY, Kimura K, Tsuji M. [Immunoelectrophoresis For Anthelmintics Evaluation Against Experimental Paragonimiasis]. Korean J Parasitol 1976;14(2):94–102.
 
2. Kagan IG, Norman L. Analysis of Helminth Antigens (Echinococcus Granulosus and Schistosoma Mansoni) by Agar Gel Methods. Ann N Y Acad Sci 1963;113:130–153.
  
3. Kent NH. Comparative Immunochemistry of Larval and Adult from of Schistosoma Mansoni. Ann N Y Acad Sci 1963;113:100–113.
  
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