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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 ,
D. Acosta ,
A. Dowd ,
S. McGonicle ,
J. Dalton  and
C. Carmona
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.
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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
Information
Journal of Helminthology , Volume 71 , Issue 4 , December 1997 , pp. 333 - 338
Copyright
Copyright © Cambridge University Press 1997

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