Discovery of novel Schistosoma japonicum antigens using a targeted protein microarray approach
- McWilliam, Hamish, Driguez, Patrick, Piedrafita, David, McManus, Donald, Meeusen, Els
- Authors: McWilliam, Hamish , Driguez, Patrick , Piedrafita, David , McManus, Donald , Meeusen, Els
- Date: 2014
- Type: Text , Journal article
- Relation: Parasites and Vectors Vol. 7, no. 1 (2014), p. 1-11
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- Description: Background: Novel vaccine candidates against Schistosoma japonicum are required, and antigens present in the vulnerable larval developmental stage are attractive targets. Post-genomic technologies are now available which can contribute to such antigen discovery. Methods. A schistosome-specific protein microarray was probed using the local antibody response against migrating larvae. Antigens were assessed for their novelty and predicted larval expression and host-exposed features. One antigen was further characterised and its sequence and structure were analysed in silico. Real-time polymerase chain reaction was used to analyse transcript expression throughout development, and immunoblotting and enzyme-linked immunosorbent assays employed to determine antigen recognition by antibody samples. Results: Several known and novel antigens were discovered, two of which showed up-regulated transcription in schistosomula. One novel antigen, termed S. japonicum Ly-6-like protein 1 (Sj-L6L-1), was further characterised and shown to share structural and sequence features with the Ly-6 protein family. It was found to be present in the worm tegument and expressed in both the larval and adult worms, but was found to be antigenic only in the lungs that the larvae migrate to and traverse. Conclusions: This study represents a novel approach to vaccine antigen discovery and may contribute to schistosome vaccine development against this important group of human and veterinary pathogens. © 2014 McWilliam et al.; licensee BioMed Central Ltd.
- Authors: McWilliam, Hamish , Driguez, Patrick , Piedrafita, David , McManus, Donald , Meeusen, Els
- Date: 2014
- Type: Text , Journal article
- Relation: Parasites and Vectors Vol. 7, no. 1 (2014), p. 1-11
- Full Text:
- Reviewed:
- Description: Background: Novel vaccine candidates against Schistosoma japonicum are required, and antigens present in the vulnerable larval developmental stage are attractive targets. Post-genomic technologies are now available which can contribute to such antigen discovery. Methods. A schistosome-specific protein microarray was probed using the local antibody response against migrating larvae. Antigens were assessed for their novelty and predicted larval expression and host-exposed features. One antigen was further characterised and its sequence and structure were analysed in silico. Real-time polymerase chain reaction was used to analyse transcript expression throughout development, and immunoblotting and enzyme-linked immunosorbent assays employed to determine antigen recognition by antibody samples. Results: Several known and novel antigens were discovered, two of which showed up-regulated transcription in schistosomula. One novel antigen, termed S. japonicum Ly-6-like protein 1 (Sj-L6L-1), was further characterised and shown to share structural and sequence features with the Ly-6 protein family. It was found to be present in the worm tegument and expressed in both the larval and adult worms, but was found to be antigenic only in the lungs that the larvae migrate to and traverse. Conclusions: This study represents a novel approach to vaccine antigen discovery and may contribute to schistosome vaccine development against this important group of human and veterinary pathogens. © 2014 McWilliam et al.; licensee BioMed Central Ltd.
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
- Hosking, Christopher, McWilliam, Hamish, Driguez, Patrick, Piedrafita, David, Li, Yuesheng, McManus, Donald, Ilag, Leodevico, Meeusen, Els, De Veer, Michael
- Authors: Hosking, Christopher , McWilliam, Hamish , Driguez, Patrick , Piedrafita, David , Li, Yuesheng , McManus, Donald , Ilag, Leodevico , Meeusen, Els , De Veer, Michael
- Date: 2015
- Type: Text , Journal article
- Relation: PLoS Neglected Tropical Diseases Vol. 9, no. 12 (2015), p. 1-20
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- Description: 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. © 2015 Hosking et al.
- Authors: Hosking, Christopher , McWilliam, Hamish , Driguez, Patrick , Piedrafita, David , Li, Yuesheng , McManus, Donald , Ilag, Leodevico , Meeusen, Els , De Veer, Michael
- Date: 2015
- Type: Text , Journal article
- Relation: PLoS Neglected Tropical Diseases Vol. 9, no. 12 (2015), p. 1-20
- Full Text:
- Reviewed:
- Description: 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. © 2015 Hosking et al.
Local immune responses of the Chinese water buffalo, Bubalus bubalis, against Schistosoma japonicum larvae: crucial insights for vaccine design
- McWilliam, Hamish, Piedrafita, David, Li, Yuesheng, Zheng, Mao, He, Yongkang, Yu, Xinling, McManus, Donald, Meeusen, Els
- Authors: McWilliam, Hamish , Piedrafita, David , Li, Yuesheng , Zheng, Mao , He, Yongkang , Yu, Xinling , McManus, Donald , Meeusen, Els
- Date: 2013
- Type: Text , Journal article
- Relation: PLoS Neglected Tropical Diseases. Vol.7, no.9(Art.No: e2460) (2013), p. 1-11
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- Description: Asian schistosomiasis is a zoonotic parasitic disease infecting up to a million people and threatening tens of millions more. Control of this disease is hindered by the animal reservoirs of the parasite, in particular the water buffalo (Bubalus bubalis), which is responsible for significant levels of human transmission. A transmission-blocking vaccine administered to buffaloes is a realistic option which would aid in the control of schistosomiasis. This will however require a better understanding of the immunobiology of schistosomiasis in naturally exposed buffaloes, particularly the immune response to migrating schistosome larvae, which are the likely targets of an anti-schistosome vaccine. To address this need we investigated the immune response at the major sites of larval migration, the skin and the lungs, in previously exposed and re-challenged water buffaloes. In the skin, a strong allergic-type inflammatory response occurred, characterised by leukocyte and eosinophil infiltration including the formation of granulocytic abscesses. Additionally at the local skin site, interleukin-5 transcript levels were elevated, while interleukin-10 levels decreased. In the skin-draining lymph node (LN) a predominant type-2 profile was seen in stimulated cells, while in contrast a type-1 profile was detected in the lung draining LN, and these responses occurred consecutively, reflecting the timing of parasite migration. The intense type-2 immune response at the site of cercarial penetration is significantly different to that seen in naive and permissive animal models such as mice, and suggests a possible mechanism for immunity. Preliminary data also suggest a reduced and delayed immune response occurred in buffaloes given high cercarial challenge doses compared with moderate infections, particularly in the skin. This study offers a deeper understanding into the immunobiology of schistosomiasis in a natural host, which may aid in the future design of more effective vaccines.
- Authors: McWilliam, Hamish , Piedrafita, David , Li, Yuesheng , Zheng, Mao , He, Yongkang , Yu, Xinling , McManus, Donald , Meeusen, Els
- Date: 2013
- Type: Text , Journal article
- Relation: PLoS Neglected Tropical Diseases. Vol.7, no.9(Art.No: e2460) (2013), p. 1-11
- Full Text:
- Reviewed:
- Description: Asian schistosomiasis is a zoonotic parasitic disease infecting up to a million people and threatening tens of millions more. Control of this disease is hindered by the animal reservoirs of the parasite, in particular the water buffalo (Bubalus bubalis), which is responsible for significant levels of human transmission. A transmission-blocking vaccine administered to buffaloes is a realistic option which would aid in the control of schistosomiasis. This will however require a better understanding of the immunobiology of schistosomiasis in naturally exposed buffaloes, particularly the immune response to migrating schistosome larvae, which are the likely targets of an anti-schistosome vaccine. To address this need we investigated the immune response at the major sites of larval migration, the skin and the lungs, in previously exposed and re-challenged water buffaloes. In the skin, a strong allergic-type inflammatory response occurred, characterised by leukocyte and eosinophil infiltration including the formation of granulocytic abscesses. Additionally at the local skin site, interleukin-5 transcript levels were elevated, while interleukin-10 levels decreased. In the skin-draining lymph node (LN) a predominant type-2 profile was seen in stimulated cells, while in contrast a type-1 profile was detected in the lung draining LN, and these responses occurred consecutively, reflecting the timing of parasite migration. The intense type-2 immune response at the site of cercarial penetration is significantly different to that seen in naive and permissive animal models such as mice, and suggests a possible mechanism for immunity. Preliminary data also suggest a reduced and delayed immune response occurred in buffaloes given high cercarial challenge doses compared with moderate infections, particularly in the skin. This study offers a deeper understanding into the immunobiology of schistosomiasis in a natural host, which may aid in the future design of more effective vaccines.
Specific humoral response of hosts with variable schistosomiasis susceptibility
- Driguez, Patrick, McWilliam, Hamish, Gaze, Soraya, Piedrafita, David, Pearson, Mark, Nakajima, Rie, Duke, Mary, Trieu, Angela, Doolan, Denise, Cardoso, Fernanda, Jasinskas, Algis, Gobert, Geoffrey, Felgner, Philip, Loukas, Alex, Meeusen, Els, McManus, Donald
- Authors: Driguez, Patrick , McWilliam, Hamish , Gaze, Soraya , Piedrafita, David , Pearson, Mark , Nakajima, Rie , Duke, Mary , Trieu, Angela , Doolan, Denise , Cardoso, Fernanda , Jasinskas, Algis , Gobert, Geoffrey , Felgner, Philip , Loukas, Alex , Meeusen, Els , McManus, Donald
- Date: 2016
- Type: Text , Journal article
- Relation: Immunology and Cell Biology Vol. 94, no. 1 (2016), p. 52-65
- Full Text:
- Reviewed:
- Description: The schistosome blood flukes are some of the largest global causes of parasitic morbidity. Further study of the specific antibody response during schistosomiasis may yield the vaccines and diagnostics needed to combat this disease. Therefore, for the purposes of antigen discovery, sera and antibody-secreting cell (ASC) probes from semi-permissive rats and sera from susceptible mice were used to screen a schistosome protein microarray. Following Schistosoma japonicum infection, rats had reduced pathology, increased antibody responses and broader antigen recognition profiles compared with mice. With successive infections, rat global serological reactivity and the number of recognized antigens increased. The local antibody response in rat skin and lung, measured with ASC probes, increased after parasite migration and contributed antigen-specific antibodies to the multivalent serological response. In addition, the temporal variation of anti-parasite serum antibodies after infection and reinfection followed patterns that appear related to the antigen driving the response. Among the 29 antigens differentially recognized by the infected hosts were numerous known vaccine candidates, drug targets and several S. japonicum homologs of human schistosomiasis resistance markers - the tegument allergen-like proteins. From this set, we prioritized eight proteins that may prove to be novel schistosome vaccine and diagnostic antigens. © 2016 Australasian Society for Immunology Inc. All rights reserved.
- Authors: Driguez, Patrick , McWilliam, Hamish , Gaze, Soraya , Piedrafita, David , Pearson, Mark , Nakajima, Rie , Duke, Mary , Trieu, Angela , Doolan, Denise , Cardoso, Fernanda , Jasinskas, Algis , Gobert, Geoffrey , Felgner, Philip , Loukas, Alex , Meeusen, Els , McManus, Donald
- Date: 2016
- Type: Text , Journal article
- Relation: Immunology and Cell Biology Vol. 94, no. 1 (2016), p. 52-65
- Full Text:
- Reviewed:
- Description: The schistosome blood flukes are some of the largest global causes of parasitic morbidity. Further study of the specific antibody response during schistosomiasis may yield the vaccines and diagnostics needed to combat this disease. Therefore, for the purposes of antigen discovery, sera and antibody-secreting cell (ASC) probes from semi-permissive rats and sera from susceptible mice were used to screen a schistosome protein microarray. Following Schistosoma japonicum infection, rats had reduced pathology, increased antibody responses and broader antigen recognition profiles compared with mice. With successive infections, rat global serological reactivity and the number of recognized antigens increased. The local antibody response in rat skin and lung, measured with ASC probes, increased after parasite migration and contributed antigen-specific antibodies to the multivalent serological response. In addition, the temporal variation of anti-parasite serum antibodies after infection and reinfection followed patterns that appear related to the antigen driving the response. Among the 29 antigens differentially recognized by the infected hosts were numerous known vaccine candidates, drug targets and several S. japonicum homologs of human schistosomiasis resistance markers - the tegument allergen-like proteins. From this set, we prioritized eight proteins that may prove to be novel schistosome vaccine and diagnostic antigens. © 2016 Australasian Society for Immunology Inc. All rights reserved.
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