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

Christopher G Hosking; Patrick Driguez; Hamish E G McWilliam; Leodevico L Ilag; Simon Gladman; Yuesheng Li; David Piedrafita; Donald P McManus; Els N T Meeusen; Michael J de Veer

doi:10.1016/j.ijpara.2015.05.002

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

Int J Parasitol. 2015 Sep;45(11):729-40. doi: 10.1016/j.ijpara.2015.05.002. Epub 2015 Jun 24.

Affiliations

¹ Biotechnology Research Laboratories, Department of Physiology, Monash University, Victoria 3800, Australia.
² QIMR Berghofer Medical Research Institute, Brisbane, Queensland 4006, Australia.
³ Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Victoria 3000, Australia.
⁴ Bio21, Molecular Sciences and Biotechnology Institute, The University of Melbourne, Victoria 3052, Australia.
⁵ Victorian Life Sciences Computation Initiative, The University of Melbourne Parkville, Victoria 3052, Australia.
⁶ School of Applied and Biomedical Science, Federation University Australia, Gippsland Campus, Churchill, Victoria 3842, Australia; Department of Microbiology, Monash University, Victoria 3800, Australia.
⁷ Department of Microbiology, Monash University, Victoria 3800, Australia.
⁸ Biotechnology Research Laboratories, Department of Physiology, Monash University, Victoria 3800, Australia. Electronic address: [email protected].

PMID: 26116907
DOI: 10.1016/j.ijpara.2015.05.002

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.

Keywords: Buffalo; China; Phage-display; Protein microarray; Schistosoma japonicum; Schistosomule; Single-chain Fv antibody; scFv.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Helminth / immunology*
Antibodies, Helminth / isolation & purification
Antigens, Helminth / immunology
Buffaloes*
Larva / growth & development
Larva / immunology
Lymph Nodes / immunology
Protein Array Analysis
Schistosoma japonicum / growth & development*
Schistosoma japonicum / immunology*
Schistosomiasis japonica / immunology*
Schistosomiasis japonica / veterinary*
Single-Chain Antibodies / immunology*
Single-Chain Antibodies / isolation & purification

Substances

Antibodies, Helminth
Antigens, Helminth
Single-Chain Antibodies