Hostname: page-component-cd9895bd7-p9bg8 Total loading time: 0 Render date: 2024-12-18T08:04:26.441Z Has data issue: false hasContentIssue false

Cysteine proteinase activity is required for survival of the parasite in experimental acute amoebic liver abscesses in hamsters

Published online by Cambridge University Press:  10 June 2004

A. OLIVOS-GARCÍA
Affiliation:
Department of Experimental Medicine, National Autonomous University of Mexico Medical School, México City, México
E. TELLO
Affiliation:
Department of Experimental Medicine, National Autonomous University of Mexico Medical School, México City, México
M. NEQUIZ-AVENDAÑO
Affiliation:
Department of Experimental Medicine, National Autonomous University of Mexico Medical School, México City, México
A. GONZÁLEZ-CANTO
Affiliation:
Department of Experimental Medicine, National Autonomous University of Mexico Medical School, México City, México
R. LÓPEZ-VANCELL
Affiliation:
Department of Experimental Medicine, National Autonomous University of Mexico Medical School, México City, México
M. C. GARCÍA DE LEÓN
Affiliation:
Department of Experimental Medicine, National Autonomous University of Mexico Medical School, México City, México
I. MONTFORT
Affiliation:
Department of Experimental Medicine, National Autonomous University of Mexico Medical School, México City, México
R. PÉREZ-TAMAYO
Affiliation:
Department of Experimental Medicine, National Autonomous University of Mexico Medical School, México City, México

Abstract

Axenic trophozoites of Entamoeba histolytica strain HM1-IMSS grown in vitro in the presence of E-64, a potent irreversible inhibitor of cysteine proteinases, preserved their viability, their rate of replication, their resistance to complement, their haemolytic capacity and their ability to destroy target cells, despite complete inhibition of total cysteine proteinase activity. On the other hand, their erythrophagocytic capacity and their ability to decrease TER of MDCK cells was partially decreased. The same trophozoites injected into the portal vein of hamsters receiving a maintaining dose of E-64 failed to cause tissue damage and were rapidly eliminated. Our results suggest that amoebic cysteine proteinase activity is not required for amoebic functions in in vitro conditions, but that it becomes necessary for survival of trophozoites in in vivo conditions, whatever other role (if any) it may play in the parasite's virulence.

Type
Research Article
Copyright
© 2004 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

ANKRI, S., STOLARSKY, T., BRACHA, R., PADILLA-VACA, F. & MIRELMAN, D. (1999). Antisense inhibition of expression of cysteine proteinases affects Entamoeba histolytica-induced formation of liver abscess in hamsters. Infection and Immunity 67, 421422.Google Scholar
ANKRI, S., STOLARSKY, T. & MIRELMAN, D. (1998). Antisense inhibition of expression of cysteine proteinases does not affect Entamoeba histolytica cytopathic or haemolytic activity but inhibits phagocytosis. Molecular Microbiology 28, 777785.CrossRefGoogle Scholar
BRUCHHAUS, I., LOFTUS, B. J., HALL, N. & TANNICH, E. (2003). The intestinal protozoan parasite Entamoeba histolytica contains 20 cysteine protease genes, of which only a small subset is expressed during in vitro cultivation. Eukaryotic Cell 2, 501509.CrossRefGoogle Scholar
DE MEESTER, F., SHAW, E., SCHOLZE, H., STOLARSKY, T. & MIRELMAN, D. (1990). Specific labeling of cysteine proteinases in pathogenic and nonpathogenic Entamoeba histolytica. Infection and Immunity 58, 13961401.Google Scholar
DIAMOND, L. S., HARLOW, D. R. & CUNNICK, C. C. (1978). A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Transactions of the Royal Society of Tropical Medicine and Hygiene 72, 431432.CrossRefGoogle Scholar
ENGEL, J. C., DOYLE, P. S., PALMER, J., HSIEH, I., BAINTON, D. F. & McKERROW, J. H. (1998). Cysteine protease inhibitors alter Golgi complex ultrastructure and function in Trypanosoma cruzi. Journal of Cell Science 111, 597606.Google Scholar
HELLBERG, A., NICKEL, R., LOTTER, H., TANNICH, E. & BRUCHHAUS, I. (2001). Overexpression of cysteine proteinase 2 in Entamoeba histolytica or Entamoeba dispar increases amoeba-induced monolayer destruction in vitro but does not augment amoebic liver abscess formation in gerbils. Cellular Microbiology 3, 1320.CrossRefGoogle Scholar
KATUNUMA, N. & KOMINAMI, E. (1995). Structure, properties, mechanisms, and assays of cysteine protease inhibitors: cystatins and E-64 derivatives. Methods in Enzymology 251, 382397.CrossRefGoogle Scholar
KEENE, W. E., PETITT, M. G., ALLEN, S. & McKERROW, J. H. (1986). The major neutral proteinase of Entamoeba histolytica. Journal of Experimental Medicine 163, 536549.CrossRefGoogle Scholar
KELLER, F., WALTER, C., LÖHDEN, U., HANKE, W., BAKKER-GRUNWALD, T. & TRISSL, D. (1988). Pathogenic and non-pathogenic Entamoeba: pore formation and hemolytic activity. Journal Protozoology 35, 359365.CrossRefGoogle Scholar
LAEMMLI, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680685.CrossRefGoogle Scholar
LI, E., YANG, W. G., ZHANG, T. & STANLEY, S. L. (1995). Interaction of laminin with Entamoeba histolytica cysteine proteinases and its effect on amebic pathogenesis. Infection and Immunity 63, 41504153.CrossRefGoogle Scholar
LÓPEZ-VANCELL, R., MONTFORT, I. & PÉREZ-TAMAYO, R. (2000). Galactose-specific adhesion and cytotoxicity of Entamoeba histolytica. Parasitology Research 86, 226231.CrossRefGoogle Scholar
LUACES, A. L. & BARRETT, A. J. (1988). Affinity purification and biochemical characterization of histolysin, the major cysteine proteinase of Entamoeba histolytica. The Biochemical Journal 250, 903909.CrossRefGoogle Scholar
LUSHBAUGH, W. B., HOFBAUER, A. F. & PITTMAN, F. E. (1985). Entamoeba histolytica: Purification of cathepsin B. Experimental Parasitology 59, 328336.CrossRefGoogle Scholar
MONTFORT, I., PÉREZ-TAMAYO, R., GONZÁLEZ, C. A., GARCÍA DE LEÓN, M. C., OLIVOS, A. & TELLO, E. (1993). Role of cysteine proteinases of Entamoeba histolytica on the cytopathogenicity of axenic trophozoites on rat and hamster hepatocytes in vitro. Journal of Parasitology 79, 98105.CrossRefGoogle Scholar
NAVARRO-GARCÍA, F., CHÁVEZ-DUEÑAS, L., TSUTSUMI, V., POSADAS DEL RIO, F. & LÓPEZ-REVILLA, R. (1995). Entamoeba histolytica: increase of enterotoxicity and of 53- and 75-kDa cysteine proteinases in a clone of higher virulence. Experimental Parasitology 80, 361372.CrossRefGoogle Scholar
NORTH, M. J., MOTTRAM, J. C. & COOMBS, G. H. (1990). Cysteine proteinases of parasitic protozoa. Parasitology Today 6, 270275.CrossRefGoogle Scholar
OLIVOS-GARCÍA, A., GONZÁLEZ-CANTO, A., LÓPEZ-VANCELL, R., GARCÍA DE LEÓN, M. C., TELLO, E., NEQUIZ-AVENDAÑO, M., MONTFORT, I. & PÉREZ-TAMAYO, R. (2003). Amebic cysteine proteinase 2 (EhCP2) plays either a minor or no role in tissue damage in acute experimental amebic liver abscess in hamsters. Parasitology Research 90, 212220.CrossRefGoogle Scholar
PÉREZ-MONTFORT, R., OSTOA-SALOMA, P., VELÁZQUEZ-MEDINA, L., MONTFORT, I. & BECKER, I. (1987). Catalytic classes of proteinases of Entamoeba histolytica. Molecular and Biochemical Parasitology 26, 8798.CrossRefGoogle Scholar
PÉREZ-TAMAYO, R., MONTFORT, I., TELLO, E. & OLIVOS, A. (1992). Ischemia in experimental acute amebic liver abscess in hamsters. International Journal for Parasitology 22, 125129.CrossRefGoogle Scholar
QUE, X. & REED, S. L. (2000). Cysteine proteinases and the pathogenesis of amebiasis. Clinical Microbiology Reviews 13, 196206.CrossRefGoogle Scholar
SAJID, M. & McKERROW, J. H. (2002). Cysteine proteases of parasitic organisms. Molecular and Biochemical Parasitology 120, 121.CrossRefGoogle Scholar
SCHOLZE, H. & TANNICH, E. (1994). Cysteine endopeptidases of Entamoeba histolytica. Methods in Enzymology 244, 512523.CrossRefGoogle Scholar
STANLEY, S. L. & REED, S. L. (2001). Microbes and microbial toxins: paradigms for microbial mucosal interactions VI. Entamoeba histolytica: parasite–host interactions. American Journal of Physiology, Gastrointestinal and Liver Physiology 280, G1049G1054.Google Scholar
STANLEY, S. L., ZHANG, T., RUBIN, D. & LI, E. (1995). Role of the Entamoeba histolytica cysteine proteinase in amebic liver abscess formation in severe combined immunodeficient mice. Infection and Immunity 63, 15871590.Google Scholar
WILCOX, D. & MASON, W. (1992). Inhibition of cysteine proteinases in lysosomes and whole cells. Biochemistry Journal 285, 495502.CrossRefGoogle Scholar
ZHANG, Z., YAN, L., WANG, L., SEYDEL, K. B., LI, E., ANKRI, S., MIRELMAN, D. & STANLEY, S. L. (2000). Entamoeba histolytica cysteine proteinases with interleukin-1 beta converting enzyme (ICE) activity cause intestinal inflammation and tissue damage in amoebiasis. Molecular Microbiology 37, 542548.CrossRefGoogle Scholar