title: Probing of a human proteome microarray with a recombinant pathogen protein reveals a novel mechanism by which hookworms suppress B-cell receptor signaling.
abstract: Na-ASP-2 is an efficacious hookworm vaccine antigen. However, despite elucidation of its crystal structure and studies addressing its immunobiology, the function of Na-ASP-2 has remained elusive. We probed a 9000-protein human proteome microarray with Na-ASP-2 and showed binding to CD79A, a component of the B-cell antigen receptor complex. Na-ASP-2 bound to human B lymphocytes ex vivo and downregulated the transcription of approximately 1000 B-cell messenger RNAs (mRNAs), while only approximately 100 mRNAs were upregulated, compared with control-treated cells. The expression of a range of molecules was affected by Na-ASP-2, including factors involved in leukocyte transendothelial migration pathways and the B-cell signaling receptor pathway. Of note was the downregulated transcription of lyn and pi3k, molecules that are known to interact with CD79A and control B-cell receptor signaling processes. Together, these results highlight a previously unknown interaction between a hookworm-secreted protein and B cells, which has implications for helminth-driven immunomodulation and vaccine development. Further, the novel use of human protein microarrays to identify host-pathogen interactions, coupled with ex vivo binding studies and subsequent analyses of global gene expression in human host cells, demonstrates a new pipeline by which to explore the molecular basis of infectious diseases.
authors.affiliation: Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns.
authors.forename: Leon
authors.initials: L
authors.lastname: Tribolet
authors.affiliation: Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns Department of Veterinary Medicine, University of Cambridge, United Kingdom.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Cantacessi
authors.affiliation: Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns.
authors.forename: Darren A
authors.initials: DA
authors.lastname: Pickering
authors.affiliation: Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns.
authors.forename: Severine
authors.initials: S
authors.lastname: Navarro
authors.affiliation: QIMR Berghofer Medical Research Institute.
authors.forename: Denise L
authors.initials: DL
authors.lastname: Doolan
authors.affiliation: QIMR Berghofer Medical Research Institute.
authors.forename: Angela
authors.initials: A
authors.lastname: Trieu
authors.affiliation: BGI-Shenzhen, China.
authors.forename: Huang
authors.initials: H
authors.lastname: Fei
authors.affiliation: BGI-Shenzhen, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Chao
authors.affiliation: Eskitis Institute, Griffith University, Brisbane.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hofmann
authors.affiliation: Faculty of Veterinary Science, University of Melbourne, Australia.
authors.forename: Robin B
authors.initials: RB
authors.lastname: Gasser
authors.affiliation: Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Giacomin
authors.affiliation: Centre for Biodiscovery and Molecular Development of Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns.
authors.forename: Alex
authors.initials: A
authors.lastname: Loukas
mesh_terms: D000328:Adult; D000723:Ancylostomatoidea / Q000276:immunology*; D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D051925:CD79 Antigens / Q000276:immunology; D002478:Cells, Cultured; D015536:Down-Regulation / Q000235:genetics / Q000276:immunology; D015801:Helminth Proteins / Q000276:immunology; D006725:Hookworm Infections / Q000276:immunology*; D054884:Host-Pathogen Interactions / Q000235:genetics / Q000276:immunology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008875:Middle Aged; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000276:immunology; D054420:Pre-B Cell Receptors / Q000276:immunology*; D040081:Protein Array Analysis / Q000379:methods; D020543:Proteome / Q000235:genetics / Q000276:immunology*; D012333:RNA, Messenger / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology*; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology*; D014158:Transcription, Genetic / Q000235:genetics / Q000276:immunology; D015854:Up-Regulation / Q000235:genetics / Q000276:immunology; D019061:src-Family Kinases / Q000235:genetics / Q000276:immunology
chemical_list: D000947:Antigens, Helminth; D051925:CD79 Antigens; D015801:Helminth Proteins; D054420:Pre-B Cell Receptors; D020543:Proteome; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019869:Phosphatidylinositol 3-Kinases; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases
keywords: B cell; CD79A; Na-ASP-2; Necator americanus; SCP/TAPS; antigen receptor; hookworm; host–pathogen interaction; protein microarray