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Amebic cysteine proteinase 2 (EhCP2) plays either a minor or no role in tissue damage in acute experimental amebic liver abscess in hamsters

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Abstract

Amebic cysteine protease 2 (EhCP2) was purified from ethyl ether extracts of axenically grown trophozoites of Entamoeba histolytica strain HM1-IMSS. The purification procedure involved molecular filtration and electroelution. Sequence analysis of the purified product revealed EhCP2 and ubiquitin(s). Electrophoretic migration patterns, isoelectric point determination and Western blot studies failed to reveal other EhCP molecules. Polyclonal antibodies against the purified EhCP2 prepared in rabbits either stabilized or enhanced the enzyme activity in a dose-response manner. Purified EhCP2 was enclosed within inert resin microspheres (22–44 μm in diameter) and injected into the portal vein of normal hamsters. In the liver, the microspheres caused mild acute inflammation and occasional minimal necrosis of short duration. Sections of the liver were immunohistochemically stained with the anti-EhCP2 antibody and the microspheres were positive for only a very short period (1 h) after injection. Sections of experimental acute (1 day, 5 days) amebic liver abscess produced in hamsters were also stained with the anti-EhCP2 antibody; and amebas were intensely positive but no staining was observed at any time in the surrounding necrotic structures. It is suggested that EhCP2 plays either a minor or no role in the causation of tissue damage in experimental acute liver amebiasis.

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Acknowledgements

We gratefully acknowledge the excellent technical help of Pedro Balderas Flores and the professional animal care of Ricardo Vargas Orozco and Daniel Sánchez Almaraz. Dr. Iris Bruchhaus kindly gave us a sample of recombinant EhCP5 prepared in her laboratory. This work includes part of the doctoral dissertation of A.O.-G. and was supported by CONACyT grants 30831-M and 25119-M and by DGAPA grant IN-225998. All experiments were conducted according to the directions of the General Health Law of Mexico.

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  1. Department of Experimental Medicine, National University of México Medical School, Ciudad Universitaria, Apartado Postal 70641, 04510, México, Districto Federal, México

    Alfonso Olivos-García, Augusto González-Canto, Rosario López-Vancell, Maria del Carmen García de León, Eusebio Tello, Mario Nequiz-Avendaño, Irmgard Montfort & Ruy Pérez-Tamayo

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  1. Alfonso Olivos-García
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  2. Augusto González-Canto
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  3. Rosario López-Vancell
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  4. Maria del Carmen García de León
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  8. Ruy Pérez-Tamayo
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Correspondence to Ruy Pérez-Tamayo.

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Olivos-García, A., González-Canto, A., López-Vancell, R. et al. Amebic cysteine proteinase 2 (EhCP2) plays either a minor or no role in tissue damage in acute experimental amebic liver abscess in hamsters. Parasitol Res 90, 212–220 (2003). https://doi.org/10.1007/s00436-002-0763-8

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