A new multi-domain member of the cystatin superfamily expressed by Fasciola hepatica

Cystatins are cysteine protease inhibitors that are widespread in the plant and animal kingdoms. Cystatins are expressed by helminth parasites that may employ these proteins to regulate parasite cysteine protease activity and to modulate host immune responses. Here, we describe the cloning of a cDNA encoding a high molecular weight protein of Fasciola hepatica that contains two domains with significant identity to the cardinal cystatin signatures and four domains with degenerated cystatin signatures. This is the first report of a multi-domain cystatin in an invertebrate species. While cystatins are divided into three evolutionary related families, our phylogenetic analysis shows that all cystatin domains within this protein, like several other helminth cystatins, belong to the cystatin family 2. The DNA region encoding the domain 4 that is the best conserved at the level of its cystatin signatures was expressed in Drosophila cells and a recombinant protein was produced and purified. This protein was a potent inhibitor of the papain and of the major cysteine protease of F. hepatica, the cathepsin L1. © 2005 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved

Neogene felsic volcanic rocks in the Hoggar province: Volcanology, geochemistry and age of the Azrou trachyte-phonolite association (Algerian Sahara)

International audienceThe Azrou volcanic district, located to the south-east to the Atakor district in the Hoggar, has a landscape is governed by a number of felsic volcanic highs and dissected mafic plateau lavas.Our new Rb-Sr age (i.e. 23.1 ± 1.6 Ma) indicates that the Azrou felsic lavas are contemporaneous with the Achkal ring complexes (Anahef region). The Azrou felsic lavas (mainly trachyte and phonolite) show remarkably homogeneous compositions both in major elements (57.5 ≤ SiO2≤ 63.1 wt%; 10.8 ≤(Na2O + K2O)≤12.4 wt%), trace elements (33.2 ≤ Th ≤ 107 ppm; 170 ≤ La≤472 ppm; 8.7<(La/Yb)N < 27.3) and radiogenic isotopes (0.703359 < 87Sr/86Sr < 0.706539; 0.512727 <143Nd/144Nd < 0.512925; 2<εNd <5.84. These data indicate that the lavas have been only very weakly contaminated by the Precambrian basement. Geodynamically, this genesis coupled with the low volume of both trachytic and phonolitic trends implies the reworking of pre-existing shear-zones allowing the rapid ascent of these small batches of magmas. This is in agreement with the general model of linear delamination along these shear zones due to the Africa-Europe convergence developed by Liégeois et al. (2005) and recently imaged by the magneto-telluric investigation of Bouzid et al. (2015)

<i>Fasciola hepatica</i> cathepsin L-like proteases: biology, function and potential in the development of first generation liver fluke vaccines.

Fasciola hepatica secretes cathepsin L proteases that facilitate the penetration of the parasite through the tissues of its host, and also participate in functions such as feeding and immune evasion. The major proteases, cathepsin L1 (FheCL1) and cathepsin L2 (FheCL2) are members of a lineage that gave rise to the human cathepsin Ls, Ks and Ss, but while they exhibit similarities in their substrate specificities to these enzymes they differ in having a wider pH range for activity and an enhanced stability at neutral pH. There are presently 13 Fasciola cathepsin L cDNAs deposited in the public databases representing a gene family of at least seven distinct members, although the temporal and spatial expression of each of these members in the developmental stage of F. hepatica remains unclear. Immunolocalisation and in situ hybridisation studies, using antibody and DNA probes, respectively, show that the vast majority of cathepsin L gene expression is carried out in the epithelial cells lining the parasite gut. Within these cells the enzyme is packaged into secretory vesicles that release their contents into the gut lumen for the purpose of degrading ingested host tissue and blood. Liver flukes also express a novel multi-domain cystatin that may be involved in the regulation of cathepsin L activity. Vaccine trials in both sheep and cattle with purified native FheCL1 and FheCL2 have shown that these enzymes can induce protection, ranging from 33 to 79%, to experimental challenge with metacercariae of F. hepatica, and very potent anti-embryonation/hatch rate effects that would block parasite transmission. In this article we review the vaccine trials carried out over the past 8 years, the role of antibody and T cell responses in mediating protection and discuss the prospects of the cathepsin Ls in the development of first generation recombinant liver fluke vaccines. © 2003 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved

Enhancement of the protective efficacy of a ROP18 vaccine against chronic toxoplasmosis by nasal route

International audienceInfection with the parasite Toxoplasma gondii causes serious public health problems and is of great economic importance worldwide. No vaccine is currently available, so the design of efficient vaccine strategies is still a topical question. In this study, we evaluated the immunoprophylactic potential of a T. gondii virulence factor, the rhoptry kinase ROP18, in a mouse model of chronic toxoplasmosis: first using a recombinant protein produced in Schneider insect cells adjuvanted with poly I:C emulsified in Montanide SV71 by a parenteral route or adjuvanted with cholera toxin by the nasal route and second using a DNA plasmid encoding ROP18 adjuvanted with GM-CSF ± IL-12 DNA. If both intranasal and subcutaneous recombinant ROP18 immunizations induced predominantly anti-ROP18 IgG1 antibodies and generated a mixed systemic Th1-/Th2-type cellular immune response characterized by the production of IFN-γ, IL-2, Il-10 and IL-5, only intranasal vaccination induced a mucosal (IgA) humoral response in intestinal washes associated with a significant brain cyst reduction (50 %) after oral challenge with T. gondii cysts. DNA immunization induced antibodies and redirected the cellular immune response toward a Th1-type response (production of IFN-γ and IL-2) but did not confer protection. These results suggest that ROP18 could be a component of a subunit vaccine against toxoplasmosis and that strategies designed to enhance mucosal protective immune responses could lead to more encouraging results