title: Secreted proteomes of different developmental stages of the gastrointestinal nematode Nippostrongylus brasiliensis.
abstract: Hookworms infect more than 700 million people worldwide and cause more morbidity than most other human parasitic infections. Nippostrongylus brasiliensis (the rat hookworm) has been used as an experimental model for human hookworm because of its similar life cycle and ease of maintenance in laboratory rodents. Adult N. brasiliensis, like the human hookworm, lives in the intestine of the host and releases excretory/secretory products (ESP), which represent the major host-parasite interface. We performed a comparative proteomic analysis of infective larval (L3) and adult worm stages of N. brasiliensis to gain insights into the molecular bases of host-parasite relationships and determine whether N. brasiliensis could indeed serve as an appropriate model for studying human hookworm infections. Proteomic data were matched to a transcriptomic database assembled from 245,874,892 Illumina reads from different developmental stages (eggs, L3, L4, and adult) of N. brasiliensis yielding∼18,426 unigenes with 39,063 possible isoform transcripts. From this analysis, 313 proteins were identified from ESPs by LC-MS/MS-52 in the L3 and 261 in the adult worm. Most of the proteins identified in the study were stage-specific (only 13 proteins were shared by both stages); in particular, two families of proteins-astacin metalloproteases and CAP-domain containing SCP/TAPS-were highly represented in both L3 and adult ESP. These protein families are present in most nematode groups, and where studied, appear to play roles in larval migration and evasion of the host's immune response. Phylogenetic analyses of defined protein families and global gene similarity analyses showed that N. brasiliensis has a greater degree of conservation with human hookworm than other model nematodes examined. These findings validate the use of N. brasiliensis as a suitable parasite for the study of human hookworm infections in a tractable animal model.
authors.affiliation: From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia;
authors.forename: Javier
authors.initials: J
authors.lastname: Sotillo
authors.affiliation: §Parasite Genomics, Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK;
authors.forename: Alejandro
authors.initials: A
authors.lastname: Sanchez-Flores
authors.affiliation: From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia; ¶Department of Veterinary Medicine, University of Cambridge, Cambridge CB3 0ES, UK;
authors.forename: Cinzia
authors.initials: C
authors.lastname: Cantacessi
authors.affiliation: ‖Institute of Immunology and Infection Research, University of Edinburgh, Ashworth Laboratories, West Mains Road, Edinburgh EH9 3JT, UK;
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Harcus
authors.affiliation: From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia;
authors.forename: Darren
authors.initials: D
authors.lastname: Pickering
authors.affiliation: From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia;
authors.forename: Tiffany
authors.initials: T
authors.lastname: Bouchery
authors.affiliation: From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia;
authors.forename: Mali
authors.initials: M
authors.lastname: Camberis
authors.affiliation: From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia;
authors.forename: Shiau-Choot
authors.initials: SC
authors.lastname: Tang
authors.affiliation: From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia;
authors.forename: Paul
authors.initials: P
authors.lastname: Giacomin
authors.affiliation: From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia; ‡‡Queensland Institute of Medical Research, Brisbane, Queensland, Australia;
authors.forename: Jason
authors.initials: J
authors.lastname: Mulvenna
authors.affiliation: §§The Genome Institute, Washington University School of Medicine, St. Louis, Missouri; ¶¶Department of Medicine, Washington University School of Medicine, St. Louis, Missouri.
authors.forename: Makedonka
authors.initials: M
authors.lastname: Mitreva
authors.affiliation: §Parasite Genomics, Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1SA, UK;
authors.forename: Matthew
authors.initials: M
authors.lastname: Berriman
authors.affiliation: From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia;
authors.forename: Graham
authors.initials: G
authors.lastname: LeGros
authors.affiliation: ‖Institute of Immunology and Infection Research, University of Edinburgh, Ashworth Laboratories, West Mains Road, Edinburgh EH9 3JT, UK;
authors.forename: Rick M
authors.initials: RM
authors.lastname: Maizels
authors.affiliation: From the ‡Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute for Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia; alex.loukas@jcu.edu.au.
authors.forename: Alex
authors.initials: A
authors.lastname: Loukas
mesh_terms: D000723:Ancylostomatoidea / Q000254:growth & development* / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D017124:Conserved Sequence; D041981:Gastrointestinal Tract / Q000469:parasitology*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D015801:Helminth Proteins / Q000378:metabolism*; D008018:Life Cycle Stages; D010802:Phylogeny; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D017423:Sequence Analysis, RNA
chemical_list: D015801:Helminth Proteins; D020543:Proteome