Hostname: page-component-7bb8b95d7b-s9k8s Total loading time: 0 Render date: 2024-09-12T03:58:04.987Z Has data issue: false hasContentIssue false

Proteinases secreted by Fasciola hepatica: time course of the inhibitory effect of serum from experimentally infected rabbits demonstrated by gelatin-substrate polyacrylamide gel electrophoresis

Published online by Cambridge University Press:  05 June 2009

L. Piacenza
Affiliation:
Unidad de Biología Parasitaria, Facultad de Ciencias, Institute de Higiene, Av. A. Navarro 3051, CP 11600, Montevideo, Uruguay:
D. Acosta
Affiliation:
Unidad de Biología Parasitaria, Facultad de Ciencias, Institute de Higiene, Av. A. Navarro 3051, CP 11600, Montevideo, Uruguay:
A. Dowd
Affiliation:
School of Biological Sciences, Dublin City University, Glasnevin, Dublin 9, Republic of Ireland
S. McGonicle
Affiliation:
School of Biological Sciences, Dublin City University, Glasnevin, Dublin 9, Republic of Ireland
J. Dalton
Affiliation:
School of Biological Sciences, Dublin City University, Glasnevin, Dublin 9, Republic of Ireland
C. Carmona*
Affiliation:
Unidad de Biología Parasitaria, Facultad de Ciencias, Institute de Higiene, Av. A. Navarro 3051, CP 11600, Montevideo, Uruguay:
*
* Author for correspondence.

Abstract

Fasciola hepatica secretes proteolytic enzymes to aid it to penetrate and migrate through the host tissues. Two of these proteinases have been purified and shown to be cathepsin L-like, and are termed, CL1 (27.5 kD) and CL2 (29 kD). The immunogenicity of these proteinases was investigated over the course of an experimental infection and following drug treatment. Four groups of rabbits were studied: group 1: orally infected with 50 metacercariae; group 2: infected and treated 8 weeks after infection; group 3: infected, treated at week 8 and reinfected at week 13 and group 4: non-infected control group. Sera were collected weekly from each group until week 20 postinfection. CL1 and CL2 were incubated with the different sera and then analysed by gelatin substrate polyacrylamide gel electrophoresis (GS-PAGE). Analysis of groups 1, 2 and 3 showed that CL1 and CL2 neutralizing antibodies appear at week 5 post-infection. In group 1, these remained throughout the 20 weeks of infection. In group 2, neutralizing antibodies disappeared at week 13, that is, 5 weeks after anti-Fasciola treatment. In group 3, CL1 and CL2 neutralizing antibodies disappeared at week 13 but reappeared by week 15, that is 2 weeks after reinfection. Pooled sera from group 4, showed no inhibitory capacity. ELISA results using CL1 and CL2 as antigen, correlate very well with the inhibitiory time course observed by GS-PAGE. These results suggest that purified cathepsin Ls are antigenic molecules recognized early in the infective process and capable of inducing a specific humoral response, strong enough to neutralize, at least partially, their enzymatic activity.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 1997

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

Araujo-Jorge, T., Lage, M., Riv, M., Carlier, Y. & van Leuven, F. (1992) Trypanosoma cruzi: enhanced alphamacroglogulin levels correlate with the resistance of BALB/cj mice to acute infection. Parasitology Research 78, 215221.CrossRefGoogle ScholarPubMed
Berasaín, P., Goñi, F., Mc.Gonicle, S., Dowd, A., Dalton, J.P., Frangione, B. & Carmona, C. (1997) Proteinases secreted by Fasciola hepatica degrade extracelullar matrix and basement membrane components. Journal of Parasitology 83,15.CrossRefGoogle Scholar
Britton, C., Knox, D.P., Canto, G., Uruquhart, G. & Kennedy, M.W. (1994) The secreted and somatic proteinases of the bovine lungworm Dictyocaulus viviparus and their inhibition by antibody from infected and vaccinated animals. Parasitology 105, 325333.CrossRefGoogle Scholar
Carmona, C., Dowd, A.J., Smith, A.M. & Dalton, J.P. (1993) Cathepsin L proteinase secreted by Fasciola hepatica in vitro prevents antibody-mediated eosinophil attachment to newly excysted juveniles. Molecular and Biochemical Parasitology 62, 918.CrossRefGoogle ScholarPubMed
Chappell, C.L., Dresden, M.H., Gryseels, B. & Deer, A.M. (1990) Antibody response to Schistosoma mansoni adult worm cysteine proteinases in infected individuals. American Journal of Tropical Medicine and Hygiene 42, 335341.CrossRefGoogle ScholarPubMed
Dalton, J.P. & Heffernan, M. (1989) Thiol proteases released in vitro by Fasciola hepatica. Molecular and Biochemical Parasitology 35, 161166.CrossRefGoogle ScholarPubMed
Dalton, J.P., Dowd, A.J. & Carmona, C. (1994) Cathepsin L proteinases secreted by the parasitic trematode Fasciola hepatica. pp. 923in Erlich, R. & Nieto, A. (Eds) Biology of parasitism. Montevideo, Uruguay, Trilce.Google Scholar
de Armas-Serra, C., Gimenez-Pardo, C., Bernardina, W. & Rodriguez-Caabeiro, F. (1995) Antibody response to a protease secreted by Trichinella spiralis muscle larvae. Parasitology Research 81, 540542.CrossRefGoogle ScholarPubMed
Dowd, A.J., McGonicle, S., Smith, A.M. & Dalton, J.P. (1994) Purification and characterization of a second cathepsin L proteinase secreted by the parasitic trematode Fasciola hepatica. European Journal of Biochemistry 223, 9198.CrossRefGoogle ScholarPubMed
El-Sahly, A.M., Adbel-Rahman, H.M. & Abdel-Wahab, M.F. (1980) Alpha-1-antitrypsin and alpha-2-macroglobulin: protease inhibitors in hepatosplenic schistosomiasis. Journal of the Egyptian Medical Association 63, 213222.Google Scholar
Isaac, L., Pereira, M., Santos, M., Sampaio, E.P., Lima, N., Lage, M. & Araujo-Jorge, T. (1990) Trypanosoma cruzi: plasma levels of alpha-2-macroglobulin during experimental murine infections with reticulotropic and myotropic strains. Parasitology Research 76, 726728.CrossRefGoogle ScholarPubMed
Knox, D.P. & Kennedy, M.W. (1988) Proteinases secreted by the parasitic larval stages of Ascaris suum and their inhibition by antibody. Molecular and Biochemical Parasitology 28, 207216.CrossRefGoogle ScholarPubMed
Redinbaugh, M.G. & Turley, R.B. (1986) Adaptation of the bicinchonic acid protein assay for use with microtiter plates and sucrose gradient fractions. Analytical Biochemistry 153, 267271.CrossRefGoogle Scholar
Rhoads, M.L. (1983) Trichinella spiralis: identification and purification of superoxide dismutase. Experimental Parasitology 56, 4145.CrossRefGoogle ScholarPubMed
Rothwell, T.L. & Merrit, G.C. (1975) Vaccination against the nematode Trichostrongylus colubriformis. II. Attempts to protect guinea pigs with worm acetylcholinesterase. International Journal for Parasitology 5, 453460.CrossRefGoogle ScholarPubMed
Sexton, J.L., Milner, A.R., Panaccio, M., Waddington, J, Wijffels, G., Chandler, D., Thompson, C., Wilson, L., Spithill, T.W., Mitchell, G.F. & Campbell, N.J. (1990) Glutathione S-transferase. Novel vaccine against Fasciola hepatica infection in sheep. Journal of Immunology 145, 39053910.CrossRefGoogle ScholarPubMed
Sharp, P.J., Smith, D., Bach, W., Wagland, B. & Cobon, G. (1991) Purified glutatione S-transferases from parasites as candidate protective antigens. International Journal for Parasitology 21, 839846.CrossRefGoogle Scholar
Smith, A.M., Dowd, A.J., McGonicle, M., Keegan, P.S., Brennan, G., Trudgett, A. & Dalton, J.P. (1993a) Purification of a cathepsin L-like proteinase secreted by adult Fasciola hepatica. Molecular and Biochemical Parasitology 62, 18.CrossRefGoogle ScholarPubMed
Smith, A.M., Dowd, A.J., Heffernan, M., Robertson, C.D. & Dalton, J.P. (1993b) Fasciola hepatica: a secreted cathepsin L-like proteinase cleaves host immunoglobulin. International Journal for Parasitology 23, 977983.CrossRefGoogle ScholarPubMed
Smith, A.M., Carmona, C., Dowd, A.J., McGonicle, S., Acosta, D. & Dalton, J.P. (1994) Neutralization of the activity of a Fasciola hepatica cathepsin L proteinase by anti-cathepsin L antibodies. Parasite Immunology 16, 325328.CrossRefGoogle ScholarPubMed