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- Title
Fasciola hepatica Serine Protease Inhibitor Family (Serpins): Purposely crafted for regulating host proteases.
- Authors
De Marco Verissimo, Carolina; Jewhurst, Heather L.; Tikhonova, Irina G.; Urbanus, Rolf T.; Maule, Aaron G.; Dalton, John P.; Cwiklinski, Krystyna
- Abstract
Serine protease inhibitors (serpins) regulate proteolytic events within diverse biological processes, including digestion, coagulation, inflammation and immune responses. The presence of serpins in Fasciola hepatica excretory-secretory products indicates that the parasite exploits these to regulate proteases encountered during its development within vertebrate hosts. Interrogation of the F. hepatica genome identified a multi-gene serpin family of seven members that has expanded by gene duplication and divergence to create an array of inhibitors with distinct specificities. We investigated the molecular properties and functions of two representatives, FhSrp1 and FhSrp2, highly expressed in the invasive newly excysted juvenile (NEJ). Consistent with marked differences in the reactive centre loop (RCL) that executes inhibitor-protease complexing, the two recombinant F. hepatica serpins displayed distinct inhibitory profiles against an array of mammalian serine proteases. In particular, rFhSrp1 efficiently inhibited kallikrein (Ki = 40 nM) whilst rFhSrp2 was a highly potent inhibitor of chymotrypsin (Ki = 0.07 nM). FhSrp1 and FhSrp2 are both expressed on the NEJ surface, predominantly around the oral and ventral suckers, suggesting that these inhibitors protect the parasites from the harmful proteolytic effects of host proteases, such as chymotrypsin, during invasion. Furthermore, the unusual inhibition of kallikrein suggests that rFhSrp1 modulates host responses such as inflammation and vascular permeability by interfering with the kallikrein-kinin system. A vaccine combination of rFhSrp1 and rFhSrp2 formulated in the adjuvant Montanide ISA 206VG elicited modest but non-significant protection against a challenge infection in a rat model, but did induce some protection against liver pathogenesis when compared to a control group and a group vaccinated with two well-studied vaccine candidates, F. hepatica cathepsin L2 and L3. This work highlights the importance of F. hepatica serpins to regulate host responses that enables parasite survival during infection and, coupled with the vaccine data, encourages future vaccine trials in ruminants. Author summary: Serpins are protease inhibitors that regulate various biological processes, including digestion, blood coagulation, inflammation and immune responses. The liver fluke, Fasciola hepatica, produces an array of inhibitors to regulate proteolytic enzymes they encounter during development within the mammalian host. In this study, we identified seven different serpins that have evolved to inhibit a range of host proteases. In particular, we characterized two representatives, FhSrp1 and FhSrp2, that we found highly expressed on the surface of the invasive newly excysted juvenile (NEJ), suggesting that they protect the parasites from harmful proteolytic effects during invasion. Contrasting inhibitory profiles were observed; while recombinant FhSrp1 inhibited kallikrein, recombinant FhSrp2 was a highly potent inhibitor of chymotrypsin. The unusual inhibition of kallikrein suggests that rFhSrp1 influences host responses such as inflammation and vascular permeability by interfering with the kallikrein-kinin system. Conversely, chymotrypsin is typically inhibited by trematode-specific serpins, implying a conserved mechanism to regulate digestive enzymes. The ability of the liver fluke serpin family to inhibit such an array of proteases highlights the importance of these inhibitors in parasite-host interactions and encourages future investigations of serpins as candidate anti-parasite vaccine targets for the control of fasciolosis in ruminants.
- Subjects
FASCIOLA hepatica; SERPINS; PROTEASE inhibitors; DIGESTIVE enzymes; HIV protease inhibitors; LIVER flukes
- Publication
PLoS Neglected Tropical Diseases, 2020, Vol 15, Issue 8, p1
- ISSN
1935-2727
- Publication type
Article
- DOI
10.1371/journal.pntd.0008510