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

Hosking, Christopher G.; Driguez, Patrick; McWilliam, Hamish E G; Ilag, Leodevico L.; Gladman, Simon; Li, Yuesheng; Piedrafita, David; McManus, Donald P.; Meeusen, Elza Nicole Theresia; Veer, Michael John de

doi:10.1016/j.ijpara.2015.05.002

Cited by 8 publications

(9 citation statements)

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“…B cells from the tumor-draining lymph node were also shown to inhibit tumors and transfer of both B cells and T cells further enhanced anticancer activity. Multiple studies have demonstrated both in humans and animals that B cells recovered from a tumor-draining lymph node are a source for antitumor antibodies [20–22]. Transfer of serum antibodies induced by a vaccine could inhibit established tumors in a mouse model [8].…”

Section: Discussionmentioning

confidence: 99%

Vaccine-draining lymph nodes of cancer patients for generating anti-cancer antibodies

et al. 2017

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BackgroundOur research is focused on using the vaccine draining lymph node to better understand the immune response to cancer vaccines and as a possible source of anti-cancer reagents. We evaluated vaccine draining lymph nodes archived from a clinical study in melanoma patients and determined the reaction of B cells to the vaccine peptides.MethodsMononuclear cells (MNCs) were recovered from cryopreserved lymph nodes that were directly receiving drainage from multi-peptide melanoma vaccine. The patients were enrolled on a vaccine study (NCT00089219, FDA, BB-IND No. 10825). B cell responses in the vaccine-draining lymph nodes were studied under both stimulated and un-stimulated conditions. Cryopreserved cells were stimulated with CD40L, stained with multiple human cell-surface markers (CD19, CD27, IgM) to identify different categories of B cell sub populations with flow cytometry. Hybridomas were generated from the lymph node cells after CD40L-stimulation. Cells were fused to murine plasmacytoma P3X63.Ag8.653 using Helix electrofusion chamber. ELISA was used to evaluate hybridoma derived antibody binding to vaccine peptides.ResultsViable MNCs were satisfactorily recovered from lymph nodes cryopreserved from six vaccine study patients 8–14 years previously. B cell ELISPOT demonstrated responses for each patient to multiple vaccine peptides. CD40L stimulation of lymph node cells increased the proportion of CD19+ CD27+ cells from 12 to 65% of the sample and increased the proportion of class-switched cells. Screening of IgG secreting clones demonstrated binding to melanoma vaccine peptides.ConclusionsB cells were successfully recovered and expanded from human cryopreserved vaccine-draining lymph nodes. Individual B cells were identified that secreted antibodies that bound to cancer vaccine peptides. The ability to reliably generate in vitro the same antibodies observed in the blood of vaccinated patients will facilitate research to understand mechanisms of human antibody activity and possibly lead to therapeutic antibodies.

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Section: Discussionmentioning

confidence: 99%

Vaccine-draining lymph nodes of cancer patients for generating anti-cancer antibodies

et al. 2017

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“…The matured buffalo antibody fragments should show little cross-reactivity with buffalo proteins and may help form the basis of a reagent to diagnose infection in buffalo. Furthermore, we have recently shown that buffalo CDRH3 regions selected for binding to schistosomules using the antibody phage display system are longer than those from non-selected libraries, suggesting that long CDRH3 regions may be important in host parasite antigen recognition [ 22 ]. Recent studies have shown that long CDRH3 regions are pivotal in the successful defence against dangerous viral infections such as HIV [ 60 ] and although the functional mechanisms for ultra-long CDRH3 regions in cattle has yet to be elucidated [ 61 ] they may offer advantages for diagnostic development.…”

Section: Discussionmentioning

confidence: 99%

“…The preparation and characterisation of a scFv-phage display library similar to that used in this study has been described previously [ 22 ]. Briefly, full length variable light (VL) and variable heavy (VH) chain genes were amplified by PCR from portal-lymph node RNA derived from buffalo 11–12 days following an experimental S .…”

Section: Methodsmentioning

confidence: 99%

“…Recently McWilliam et al, demonstrated, in a rat model of schistosomiasis, that the developing schistosome worm can elicit a distinct immune response in discrete tissue sites [ 21 ]. Building on this concept we previously published the construction of an scFv-phage library for the detection of larval stage antigens as potential vaccine candidates [ 22 ]. However, the S .…”

Section: Introductionmentioning

confidence: 99%

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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

Hosking¹,

McWilliam²,

Driguez³

et al. 2015

PLoS Negl Trop Dis

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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.

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“…The eggs lodged in the colon submucosa and mucous layers, prior to rupturing into the lumen of the bowel and passing out with the feces, can be used for clinical diagnosis. In China, field and clinical diagnosis techniques for S. japonicum are mainly dependent on stool examination of eggs and immunological detection of specific antibodies [6, 7]. Methods of schistosomiasis diagnosis using the detection of eggs include the Kato-Katz thick smear technique, a nylon screen method, the miracidium hatching test and rectal biopsy [8].…”

Section: Introductionmentioning

confidence: 99%

Clinical diagnostic value of viable Schistosoma japonicum eggs detected in host tissues

Driguez

et al. 2017

BMC Infect Dis

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Background Schistosomiasis, one of the neglected tropical diseases, is endemic in more than 70 countries. However, the clinical diagnosis of patients with a low degree of infection is an unsolved technical problem. In areas endemic for schistosomiasis japonica, proctoscopy detection of eggs has been one method used for clinical diagnosis. However, it is often a challenge to find typical live eggs and it is difficult to distinguish live eggs from large numbers of partially degraded and/or completely degraded eggs within colon biopsy tissue. To address this problem, we tested six different morphological and biochemical/molecular markers (ALP; morphological characteristics of egg; CalS (calcified substance); AOS (antioxidase); SDHG (succinic dehydrogenase) and SjR2 mRNA (retrotransposons 2 of S.japonicum genome mRNA)), including four new markers (CalS; AOS; SDHG and SjR2 mRNA.), to determine the viability of S. japonicum eggs deposited in human and mouse colon tissues. Our ultimate aim is to obtain a new method that is more sensitive, practical and accurate to clinically diagnose schistosomiasis.MethodsTissue samples were collected from mice at six different time points during S. japonicum infection with or without treatment with praziquantel (PZQ). Four new biochemical or molecular markers were used for the detection of egg viability from mouse liver and intestinal samples: CalS; AOS; SDHG and SjR2 mRNA. Subsequently, all markers were employed for the detection and analysis of eggs deposited in biopsy materials from patients with suspected schistosomiasis japonica for clinical evaluation. Microscopic examination of the egg morphology, worm burden in vivo and ALP (alkaline phosphatase) levels were used as a reference standard to evaluate the sensitivity and reliability of four new markers detecting egg viability.ResultsThe results of the study showed that the morphology of S. japonicum eggs deposited in tissues of hosts with schistosomiasis, especially cases with chronic schistosomiasis, is complex and egg viability is difficult to judge morphologically, particularly eggs with a fuzzy structure or partially modified eggs. We found that the majority of the viable schistosome eggs determined by four new markers (CalS, AOS, SDHG and SjR2 mRNA) were morphologically difficult to identify.ConclusionsAmong the markers, the most sensitive and specific method was the detection of SjR2 mRNA and the most simple, rapid and practical method was the detection of SDHG. Therefore, the detection of SDHG is the most practical for clinical application and its use could improve the accuracy in diagnosing active schistosome infection.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-017-2362-4) contains supplementary material, which is available to authorized users.

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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

Cited by 8 publications

References 58 publications

Vaccine-draining lymph nodes of cancer patients for generating anti-cancer antibodies

Vaccine-draining lymph nodes of cancer patients for generating anti-cancer antibodies

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

Clinical diagnostic value of viable Schistosoma japonicum eggs detected in host tissues

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