title: Cathepsin B homologue at the interface between a parasitic nematode and its intermediate host.
abstract: Parelaphostrongylus tenuis is a parasitic nematode that causes a debilitating neurologic disease in many North American cervids and domestic livestock species. We produced a PCR-based cDNA library from infective larvae (L3) in order to identify molecules that mediate parasitism. A dominant 1,250-bp amplicon encoded a homologue of cathepsin B cysteine proteases. The sequence incorporated a C29G substitution in the putative active site. Antibodies generated against a recombinant form detected the native protein (PtCPR-1) in Western blot assays of L3, but not adult worm, extracts. Immunohistochemical methods revealed that PtCPR-1 synthesis was restricted to larval stages within the snail intermediate host (Triodopsis sp.), beginning as early as 2 days postinfection (dpi) of snails. The protein was present in the intestine and luminal contents and was lost from larvae over time. Concurrent studies showed that larvae induced an immune response in snails beginning at 1 dpi. Layers of hemocytes encapsulated larvae immediately after infection, and granuloma-like structures formed around parasites in chronic infections. Loss of PtCPR-1 from L3 and its accumulation in host tissues coincided with degeneration of granuloma architecture 90 to 105 dpi. Fully developed L3 emerged from the snail at this time. Our data implicate PtCPR-1 in larval development and possibly in the emergence of P. tenuis from the intermediate host. Emerged L3 survived desiccation and cold stress, suggesting that they could remain infectious in the environment. Molecules promoting emergence would facilitate dispersal of L3 and increase the likelihood of transmission to definitive hosts.
authors.affiliation: James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Duffy
authors.forename: Deanne K
authors.initials: DK
authors.lastname: Cevasco
authors.forename: Dante S
authors.initials: DS
authors.lastname: Zarlenga
authors.forename: Woraporn
authors.initials: W
authors.lastname: Sukhumavasi
authors.forename: Judith A
authors.initials: JA
authors.lastname: Appleton
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002401:Cathepsin B / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D006790:Host-Parasite Interactions; D007814:Larva / Q000201:enzymology; D008681:Metastrongyloidea / Q000201:enzymology / Q000254:growth & development* / Q000472:pathogenicity*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D012908:Snails / Q000469:parasitology*
chemical_list: D002401:Cathepsin B