title: Using the local immune response from the natural buffalo host to generate an antibody fragment library that binds the early larval stages of Schistosoma japonicum.
abstract: Antibodies isolated from the local draining inguinal lymph node of field exposed-water buffaloes following challenge with Schistosoma japonicum cercariae showed high reactivity towards S. japonicum antigen preparations and bound specifically to formaldehyde-fixed S. japonicum schistosomules. Using this specific local immune response we produced a series of single-chain antibody Fv domain libraries from the same lymph nodes. Removal of phage that cross reacted with epitopes on adult parasites yielded a single-chain antibody Fv domain-phage library that specifically bound to whole formaldehyde-fixed and live S. japonicum schistosomules. DNA sequencing indicated clear enrichment of the single-chain antibody Fv domain library for buffalo B-cell complementarity determining regions post-selection for schistosomule binding. This study also revealed that long heavy chain complementarity determining regions appear to be an important factor when selecting for antibody binding fragments against schistosomule proteins. The selected single-chain antibody Fv domain-phage were used to probe a schistosome-specific protein microarray, which resulted in the recognition of many proteins expressed across all schistosome life-cycle stages. Following absorption to adult worms, the single-chain antibody Fv domain-phage library showed significantly reduced binding to most proteins, whilst two proteins (NCBI GenBank accession numbers AY915878 and AY815196) showed increased binding. We have thus developed a unique set of host derived single-chain antibody Fv domains comprising buffalo B-cell variable regions that specifically bind to early S. japonicum life-stages.
authors.affiliation: Biotechnology Research Laboratories, Department of Physiology, Monash University, Victoria 3800, Australia.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Hosking
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, Queensland 4006, Australia.
authors.forename: Patrick
authors.initials: P
authors.lastname: Driguez
authors.affiliation: Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Victoria 3000, Australia.
authors.forename: Hamish E G
authors.initials: HE
authors.lastname: McWilliam
authors.affiliation: Bio21, Molecular Sciences and Biotechnology Institute, The University of Melbourne, Victoria 3052, Australia.
authors.forename: Leodevico L
authors.initials: LL
authors.lastname: Ilag
authors.affiliation: Victorian Life Sciences Computation Initiative, The University of Melbourne Parkville, Victoria 3052, Australia.
authors.forename: Simon
authors.initials: S
authors.lastname: Gladman
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, Queensland 4006, Australia.
authors.forename: Yuesheng
authors.initials: Y
authors.lastname: Li
authors.affiliation: School of Applied and Biomedical Science, Federation University Australia, Gippsland Campus, Churchill, Victoria 3842, Australia; Department of Microbiology, Monash University, Victoria 3800, Australia.
authors.forename: David
authors.initials: D
authors.lastname: Piedrafita
authors.affiliation: QIMR Berghofer Medical Research Institute, Brisbane, Queensland 4006, Australia.
authors.forename: Donald P
authors.initials: DP
authors.lastname: McManus
authors.affiliation: Department of Microbiology, Monash University, Victoria 3800, Australia.
authors.forename: Els N T
authors.initials: EN
authors.lastname: Meeusen
authors.affiliation: Biotechnology Research Laboratories, Department of Physiology, Monash University, Victoria 3800, Australia. Electronic address: michael.deveer@monash.edu.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: de Veer
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology* / Q000302:isolation & purification; D000947:Antigens, Helminth / Q000276:immunology; D002020:Buffaloes; D007814:Larva / Q000254:growth & development / Q000276:immunology; D008198:Lymph Nodes / Q000276:immunology; D040081:Protein Array Analysis; D012549:Schistosoma japonicum / Q000254:growth & development* / Q000276:immunology*; D012554:Schistosomiasis japonica / Q000276:immunology* / Q000662:veterinary*; D057127:Single-Chain Antibodies / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D057127:Single-Chain Antibodies
keywords: Buffalo; China; Phage-display; Protein microarray; Schistosoma japonicum; Schistosomule; Single-chain Fv antibody; scFv