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Korean J Parasitol > Volume 31(2):1993 > Article

Original Article
Korean J Parasitol. 1993 Jun;31(2):117-127. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1993.31.2.117
Copyright © 1993 by The Korean Society for Parasitology
Proteinase activity in the isolates of Trichomonas vaginalis according to their pathogenicity
Y K Shim,1K H Park,1P R Chung,2 and K I Im*1
1Department of Parasitology, College of Medicine & Institute of Tropical Medicine, Yonsei University, Seoul 120-752, Korea.
Received January 14, 1993; Accepted March 02, 1993.

Abstract

Ten axenic isolates of Trichomonas vaginalis were subcutaneously injected to the BALB/c mice in order to assess their pathogenicity by means of so-called "mouse assay" method. All the isolates revealed neutral and acid proteinase activities both in their lysates and in culture media, but the specific activities of both proteinases in the severely pathogenic group were significantly higher than the mildly pathogenic group (p < 0.05). In the SDS-PAGE system in which the electrophoretic gels contained 0.4% gelatin as the substrate, five different banding patterns of trichomonal proteinases were detected, and the patterns were closely related with the pathogenicity of the isolates of T. vaginalis. All five bands might be regarded as cysteine proteinases group in the inhibitor assays. The cytotoxicity of the lysates of T. vaginalis to the target Chinese hamster ovarian (CHO) cell line was also significantly different according to the pathogenicity of the isolates, and generally lower in the lysates treated with cysteine proteinase inhibitors than in the control lysates. In summarizing the results, it might be considered that the proteinases of T. vaginalis showing five electrophoretic banding patterns are closely related with the pathogenicity and cytotoxicity of the isolates of T. vaginalis.

Figures


Fig. 1
Confidence intervals (5% significance level) of mean areas of 6-day subcutaneous lesions in mice caused by Trichomonas vaginalis inoculation.


Fig. 2
Specific activities of neutral proteinase in the mild-, moderate-, severe-pathogenic groups of Trichomonas vaginalis.


Fig. 3
Specific activities of acid proteinase in the mild-, moderate-, severe-pathogenic groups of Trichomonas vaginalis.


Fig. 4
Diagrammatic representation of proteinase banding patterns of Trichomonas vaginalis.


Fig. 5
Sodium dodesyl sulfate polyacrylamide gel electrophoresis banding patterns of proteinases in Trichomonas vaginalis.


Fig. 6
The densitometrics scan of Trichomonas vaginalis proteinase banding patterns is shown. Gels were incubated at pH 5.5 in the presense of 1 mmol dithiothreitol. Samples were run in the direction cathode (top) to anode (bottom).


Fig. 7
Cytotoxicity of Trichomonas vaginalis lysates to the Chinese hamster ovary cell line.


Fig. 8
Mean cytotoxicities of the pathogenic groups of Trichomonas vaginalis lysate to Chinese hamster ovary cell line.

Tables


Table 1
Medical history of patients from whom Trichomonas vaginalis were isolateda)


Table 2
Neutral and acid proteinase activities in Trichomonas vaginalis lysate and cultured mediaa)


Table 3
The effect of inhibitors on electrophoretic banding patterns of Trichomonas vaginalis proteinasesa)


Table 4
Effect of proteinase inhibitors on the cytotoxicity of Trichomonas vaginalis lysate

References
5. Alderete JF, Pearlman E. Pathogenic Trichomonas vaginalis cytotoxicity to cell culture monolayers. Br J Vener Dis 1984;60(2):99–105.
 
6. Alfieri SC, et al. Exp Parasit 1989;98:423–431.
 
7. Asch HL, Dresden MH. Acidic thiol proteinase activity of Schistosoma mansoni egg extracts. J Parasitol 1979;65(4):543–549.
  
8. Coombs GH. Proteinases of Leishmania mexicana and other flagellate protozoa. Parasitology 1982;84(1):149–155.
  
9. Coombs GH, North MJ. An analysis of the proteinases of Trichomonas vaginalis by polyacrylamide gel electrophoresis. Parasitology 1983;86(Pt 1):1–6.
  
10. Diamond LS. Techniques of axenic cultivation of Entamoeba histolytica Schaudinn, 1903 and E. histolytica-like amebae. J Parasitol 1968;54(5):1047–1056.
  
11. Dluzewski AR, Rangachari K, Wilson RJ, Gratzer WB. Plasmodium falciparum: protease inhibitors and inhibition of erythrocyte invasion. Exp Parasitol 1986;62(3):416–422.
  
12. Dougherty TJ, Gomer CJ, Weishaupt KR. Energetics and efficiency of photoinactivation of murine tumor cells containing hematoporphyrin. Cancer Res 1976;36(7 PT 1):2330–2333.
 
13. Fouts AC, Kraus SJ. Trichomonas vaginalis: reevaluation of its clinical presentation and laboratory diagnosis. J Infect Dis 1980;141(2):137–143.
  
14. Fulford DE, Marciano-Cabral F. Cytolytic activity of Naegleria fowleri cell-free extract. J Protozool 1986;33(4):498–502.
 
15. Gillies RJ, Didier N, Denton M. Determination of cell number in monolayer cultures. Anal Biochem 1986;159(1):109–113.
  
16. HARTLEY BS. Proteolytic enzymes. Annu Rev Biochem 1960;29:45–72.
  
17. Heath JP. Br J Vener Dis 1981;57:106–117.
 
18. Honigberg BM, Livingston MC, Frost JK. Pathogenicity of fresh isolates of Trichomonas vaginalis: "the mouse assay" versus clinical and pathologic findings. Acta Cytol 1966;10(5):353–361.
 
19. Kulda J, Honigberg BM, Frost JK, Hollander DH. Pathogenicity of Trichomonas vaginalis. a clinical and biologic study. Am J Obstet Gynecol 1970;108(6):908–918.
 
20. Laemmli UV. Nature 1970;227:680–683.
  
21. Lockwood BC, North MJ, Scott KI, Bremner AF, Coombs GH. The use of a highly sensitive electrophoretic method to compare the proteinases of trichomonads. Mol Biochem Parasitol 1987;24(1):89–95.
  
22. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(1):265–275.
 
23. Lushbaugh WB, Hofbauer AF, Pittman FE. Entamoeba histolytica: purification of cathepsin B. Exp Parasitol 1985;59(3):328–336.
  
24. Maki J, Furuhashi A, Yanagisawa T. The activity of acid proteases hydrolysing haemoglobin in parasitic helminths with special reference to interspecific and intraspecific distribution. Parasitology 1982;84(1):137–147.
  
25. McDonald JK. An overview of protease specificity and catalytic mechanisms: aspects related to nomenclature and classification. Histochem J 1985;17(7):773–785.
  
26. McLaughlin J, Faubert G. Partial purification and some properties of a neutral sulfhydryl and an acid proteinase from Entamoeba histolytica. Can J Microbiol 1977;23(4):420–425.
  
27. McLaughlin J, Muller M. Purification and characterization of a low molecular weight thiol proteinase from the flagellate protozoon Tritrichomonas foetus. J Biol Chem 1979;254(5):1526–1533.
 
28. North MJ. Comparative biochemistry of the proteinases of eucaryotic microorganisms. Microbiol Rev 1982;46(3):308–340.
 
29. Ravdin JI. Pathogenesis of disease caused by Entamoeba histolytica: studies of adherence, secreted toxins, and contact-dependent cytolysis. Rev Infect Dis 1986;8(2):247–260.
  
30. Root RK, Metcalf J, Oshino N, Chance B. H2O2 release from human granulocytes during phagocytosis. I. Documentation, quantitation, and some regulating factors. J Clin Invest 1975;55(5):945–955.
  
31. Soh CT, et al. Yonsei Med J 1961;2:31–41.
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