Research Article: Generation of a Novel Bacteriophage Library Displaying scFv Antibody Fragments from the Natural Buffalo Host to Identify Antigens from Adult Schistosoma japonicum for Diagnostic Development

Date Published: December 18, 2015

Publisher: Public Library of Science

Author(s): Christopher G. Hosking, Hamish E. G. McWilliam, Patrick Driguez, David Piedrafita, Yuesheng Li, Donald P. McManus, Leodevico L. Ilag, Els N. T. Meeusen, Michael J. de Veer, John Pius Dalton. http://doi.org/10.1371/journal.pntd.0004280

Abstract: The development of effective diagnostic tools will be essential in the continuing fight to reduce schistosome infection; however, the diagnostic tests available to date are generally laborious and difficult to implement in current parasite control strategies. We generated a series of single-chain antibody Fv domain (scFv) phage display libraries from the portal lymph node of field exposed water buffaloes, Bubalus bubalis, 11–12 days post challenge with Schistosoma japonicum cercariae. The selected scFv-phages showed clear enrichment towards adult schistosomes and excretory-secretory (ES) proteins by immunofluorescence, ELISA and western blot analysis. The enriched libraries were used to probe a schistosome specific protein microarray resulting in the recognition of a number of proteins, five of which were specific to schistosomes, with RNA expression predominantly in the adult life-stage based on interrogation of schistosome expressed sequence tags (EST). As the libraries were enriched by panning against ES products, these antigens may be excreted or secreted into the host vasculature and hence may make good targets for a diagnostic assay. Further selection of the scFv library against infected mouse sera identified five soluble scFv clones that could selectively recognise soluble whole adult preparations (SWAP) relative to an irrelevant protein control (ovalbumin). Furthermore, two of the identified scFv clones also selectively recognised SWAP proteins when spiked into naïve mouse sera. These host B-cell derived scFvs that specifically bind to schistosome protein preparations will be valuable reagents for further development of a cost effective point-of-care diagnostic test.

Partial Text: Schistosomiasis is one of the most insidious of all the tropical parasitic infections and threatens the health of hundreds of millions of people worldwide [1]. The last 20 years has seen remarkable progress in disease control through the use of praziquantel (PZQ), but this drug does not protect against re-infection and mass drug administration programmes based around its use are probably untenable long term [2–4]. Recently there has been a major focus on the development of anti-schistosome vaccines, but to date a protective commercial vaccine remains elusive [1]. As mass drug administration decreases worm burdens within endemic areas, the need for improved diagnostic tests should be given research priority [5]. However, in countries where elimination of schistosomiasis has been given precedence, case detection of infected individuals remains problematic as the commonly used methods for diagnosis lack the necessary sensitivity and specificity to accurately determine parasite burden [6]. Although application of modern research laboratory techniques has seen improvements in the diagnosis of helminth infection, uptake has not been uniform and proof of concept studies that show promise have often not been followed through with much needed product development [7].

There has been considerable success at reducing transmission and infection rates of S. japonicum in the PR China [42], however, there is a need to ensure the parasite life cycle is completely broken and that the national elimination program does not falter due to a significant number of low-level infections not being detected [11]. In areas where schistosomiasis japonica remains a problem there is a need to measure and target treatments as well as to educate communities [43]. If a significant number of low-level infections go undetected, there is the risk that the efforts already employed to control transmission will be in vain [11].

Source:

http://doi.org/10.1371/journal.pntd.0004280

 

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