Molecular characterisation and vaccine efficacy of two novel developmentally regulated surface tegument proteins of Fasciola hepatica

Paul McCusker; Hayley Toet; Vignesh Rathinasamy; Neil Young; Travis Beddoe; Glenn Anderson; Robert Dempster; Paul McVeigh; Erin McCammick; Duncan Wells; Angela Mousley; Nikki J. Marks; Aaron G. Maule; Terry W. Spithill

doi:10.1016/j.vetpar.2020.109244

Molecular characterisation and vaccine efficacy of two novel developmentally regulated surface tegument proteins of Fasciola hepatica

Paul McCusker, Hayley Toet, Vignesh Rathinasamy, Neil Young, Travis Beddoe, Glenn Anderson, Robert Dempster, Paul McVeigh, Erin McCammick, Duncan Wells, Angela Mousley, Nikki J. Marks, Aaron G. Maule, Terry W. Spithill^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

The surface tegument of Fasciola hepatica is a crucial tissue due to its key role at the host-parasite interface. We characterised three novel proteins, termed Fhteg1, Fhteg5 and Fhteg8, that are found in the tegument membrane fraction of adult F. hepatica. Bioinformatic analysis of proteomic datasets identified Fhteg5 and Fhteg8 as tegumental glycoproteins and revealed that Fhteg1, Fhteg5 and Fhteg8 are associated with exosomes of adult F. hepatica. Fhteg1, Fhteg5 and Fhteg8 appear to be related to uncharacterised sequences in F. gigantica, Fasciolopsis buski, Echinostoma caproni, Clonorchis sinensis, Opisthorchis viverrini, Schistosoma japonicum and S. mansoni, although F. hepatica appears to have expanded this family. Fhteg1 and Fhteg5 were characterised in detail. The Fhteg1 and Fhteg5 gene transcripts each demonstrate significant upregulation in juvenile fluke 2–4 days post-excystment, with transcript levels maintained during development over 3 weeks in vitro. RNAseq data showed that both Fhtegs are expressed in the adult life stage, although the transcript levels were about 8 fold lower than those in juveniles (3 week post infection). Using immunocytochemistry, Fhteg1 and Fhteg5 were each shown to be expressed in cells adjacent to the muscle layer as well as on the surface of 1 week old juveniles, whilst Fhteg5 was also present in cells at the base of the pharynx. RNAi mediated knockdown of Fhteg1 and Fhteg5 transcripts in 4–10 day old juveniles had no effect on parasite survival, movement or growth in vitro. Although no IgG responses were observed for Fhteg1 or Fhteg5 during infection in sheep and cattle, both proteins elicited a low IgG response in a proportion of infected rats. Rats vaccinated with Fhteg1 and Fhteg5 showed good IgG responses to both proteins and a mean 48.2 % reduction in worm burden following parasite challenge. Although vaccination of cattle with both proteins induced a range of IgG responses, no protection was observed against parasite challenge. This is the first study to provide insights into the molecular properties of two novel, developmentally regulated surface tegument proteins in F. hepatica.

Original language	English
Article number	109244
Journal	Veterinary Parasitology
Volume	286
Early online date	12 Sep 2020
DOIs	https://doi.org/10.1016/j.vetpar.2020.109244
Publication status	Published - Oct 2020

Bibliographical note

Funding Information:
The work in the T.W.S. laboratory was supported by funds from Linkage Project LP130100943 ?Development of a novel vaccine targeting parasite tegument proteins for liver fluke disease in livestock? and Linkage Project LP160100442 ?Optimisation of a novel hybrid vaccine for liver fluke disease in cattle? funded by the Australian Research Council, in collaboration with Virbac (Australia) Pty. Ltd. In addition, support was obtained from La Trobe University, Melbourne, Australia. V.R. was supported by a Victorian-India Doctoral Scholarship from the Australia India Institute, a PhD scholarship from La Trobe University and a La Trobe University School of Life Sciences Postgraduate Publication Award. T.B. was supported by Pfizer Australia Research Fellowship. Funding from the National Health Medical Research Council (NHMRC) of Australia is gratefully acknowledged (N.D.Y.). Work at the A.G.M. lab in QUB was supported by the Professor John Glover Memorial Fund awarded to P.M. and the National Centre for the 3Rs grant NC/N001486/1. PM received an Australian Bicentennial Scholarship. We thank Prof Mike Stear for statistical analysis and Dr Wynne Gibbison (Virbac Australia) for oversight and management of the cattle vaccine trial.

Funding Information:
The work in the T.W.S. laboratory was supported by funds from Linkage Project LP130100943 “Development of a novel vaccine targeting parasite tegument proteins for liver fluke disease in livestock” and Linkage Project LP160100442 “Optimisation of a novel hybrid vaccine for liver fluke disease in cattle” funded by the Australian Research Council, in collaboration with Virbac (Australia) Pty. Ltd. . In addition, support was obtained from La Trobe University, Melbourne, Australia. V.R. was supported by a Victorian-India Doctoral Scholarship from the Australia India Institute , a PhD scholarship from La Trobe University and a La Trobe University School of Life Sciences Postgraduate Publication Award . T.B. was supported by Pfizer Australia Research Fellowship . Funding from the National Health Medical Research Council (NHMRC) of Australia is gratefully acknowledged (N.D.Y.). Work at the A.G.M. lab in QUB was supported by the Professor John Glover Memorial Fund awarded to P.M. and the National Centre for the 3Rs grant NC/N001486/1 . PM received an Australian Bicentennial Scholarship . We thank Prof Mike Stear for statistical analysis and Dr Wynne Gibbison (Virbac Australia) for oversight and management of the cattle vaccine trial.

Publisher Copyright:
© 2020 Elsevier B.V.

Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.

Keywords

Cattle
Exosome
Fasciola hepatica
Juvenile fluke
qPCR
Rat
RNAi
Tegument membrane proteins
Vaccine

ASJC Scopus subject areas

Parasitology
veterinary(all)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Cite this

McCusker, P., Toet, H., Rathinasamy, V., Young, N., Beddoe, T., Anderson, G., Dempster, R., McVeigh, P., McCammick, E., Wells, D., Mousley, A., Marks, N. J., Maule, A. G., & Spithill, T. W. (2020). Molecular characterisation and vaccine efficacy of two novel developmentally regulated surface tegument proteins of Fasciola hepatica. Veterinary Parasitology, 286, [109244]. https://doi.org/10.1016/j.vetpar.2020.109244

@article{3a36543cf43843c1960ef684afdf6f7e,

title = "Molecular characterisation and vaccine efficacy of two novel developmentally regulated surface tegument proteins of Fasciola hepatica",

abstract = "The surface tegument of Fasciola hepatica is a crucial tissue due to its key role at the host-parasite interface. We characterised three novel proteins, termed Fhteg1, Fhteg5 and Fhteg8, that are found in the tegument membrane fraction of adult F. hepatica. Bioinformatic analysis of proteomic datasets identified Fhteg5 and Fhteg8 as tegumental glycoproteins and revealed that Fhteg1, Fhteg5 and Fhteg8 are associated with exosomes of adult F. hepatica. Fhteg1, Fhteg5 and Fhteg8 appear to be related to uncharacterised sequences in F. gigantica, Fasciolopsis buski, Echinostoma caproni, Clonorchis sinensis, Opisthorchis viverrini, Schistosoma japonicum and S. mansoni, although F. hepatica appears to have expanded this family. Fhteg1 and Fhteg5 were characterised in detail. The Fhteg1 and Fhteg5 gene transcripts each demonstrate significant upregulation in juvenile fluke 2–4 days post-excystment, with transcript levels maintained during development over 3 weeks in vitro. RNAseq data showed that both Fhtegs are expressed in the adult life stage, although the transcript levels were about 8 fold lower than those in juveniles (3 week post infection). Using immunocytochemistry, Fhteg1 and Fhteg5 were each shown to be expressed in cells adjacent to the muscle layer as well as on the surface of 1 week old juveniles, whilst Fhteg5 was also present in cells at the base of the pharynx. RNAi mediated knockdown of Fhteg1 and Fhteg5 transcripts in 4–10 day old juveniles had no effect on parasite survival, movement or growth in vitro. Although no IgG responses were observed for Fhteg1 or Fhteg5 during infection in sheep and cattle, both proteins elicited a low IgG response in a proportion of infected rats. Rats vaccinated with Fhteg1 and Fhteg5 showed good IgG responses to both proteins and a mean 48.2 % reduction in worm burden following parasite challenge. Although vaccination of cattle with both proteins induced a range of IgG responses, no protection was observed against parasite challenge. This is the first study to provide insights into the molecular properties of two novel, developmentally regulated surface tegument proteins in F. hepatica.",

keywords = "Cattle, Exosome, Fasciola hepatica, Juvenile fluke, qPCR, Rat, RNAi, Tegument membrane proteins, Vaccine",

author = "Paul McCusker and Hayley Toet and Vignesh Rathinasamy and Neil Young and Travis Beddoe and Glenn Anderson and Robert Dempster and Paul McVeigh and Erin McCammick and Duncan Wells and Angela Mousley and Marks, {Nikki J.} and Maule, {Aaron G.} and Spithill, {Terry W.}",

note = "Funding Information: The work in the T.W.S. laboratory was supported by funds from Linkage Project LP130100943 ?Development of a novel vaccine targeting parasite tegument proteins for liver fluke disease in livestock? and Linkage Project LP160100442 ?Optimisation of a novel hybrid vaccine for liver fluke disease in cattle? funded by the Australian Research Council, in collaboration with Virbac (Australia) Pty. Ltd. In addition, support was obtained from La Trobe University, Melbourne, Australia. V.R. was supported by a Victorian-India Doctoral Scholarship from the Australia India Institute, a PhD scholarship from La Trobe University and a La Trobe University School of Life Sciences Postgraduate Publication Award. T.B. was supported by Pfizer Australia Research Fellowship. Funding from the National Health Medical Research Council (NHMRC) of Australia is gratefully acknowledged (N.D.Y.). Work at the A.G.M. lab in QUB was supported by the Professor John Glover Memorial Fund awarded to P.M. and the National Centre for the 3Rs grant NC/N001486/1. PM received an Australian Bicentennial Scholarship. We thank Prof Mike Stear for statistical analysis and Dr Wynne Gibbison (Virbac Australia) for oversight and management of the cattle vaccine trial. Funding Information: The work in the T.W.S. laboratory was supported by funds from Linkage Project LP130100943 “Development of a novel vaccine targeting parasite tegument proteins for liver fluke disease in livestock” and Linkage Project LP160100442 “Optimisation of a novel hybrid vaccine for liver fluke disease in cattle” funded by the Australian Research Council, in collaboration with Virbac (Australia) Pty. Ltd. . In addition, support was obtained from La Trobe University, Melbourne, Australia. V.R. was supported by a Victorian-India Doctoral Scholarship from the Australia India Institute , a PhD scholarship from La Trobe University and a La Trobe University School of Life Sciences Postgraduate Publication Award . T.B. was supported by Pfizer Australia Research Fellowship . Funding from the National Health Medical Research Council (NHMRC) of Australia is gratefully acknowledged (N.D.Y.). Work at the A.G.M. lab in QUB was supported by the Professor John Glover Memorial Fund awarded to P.M. and the National Centre for the 3Rs grant NC/N001486/1 . PM received an Australian Bicentennial Scholarship . We thank Prof Mike Stear for statistical analysis and Dr Wynne Gibbison (Virbac Australia) for oversight and management of the cattle vaccine trial. Publisher Copyright: {\textcopyright} 2020 Elsevier B.V. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

year = "2020",

month = oct,

doi = "10.1016/j.vetpar.2020.109244",

language = "English",

volume = "286",

journal = "Veterinary Parasitology",

issn = "0304-4017",

publisher = "Elsevier",

}

McCusker, P, Toet, H, Rathinasamy, V, Young, N, Beddoe, T, Anderson, G, Dempster, R, McVeigh, P, McCammick, E, Wells, D , Mousley, A , Marks, NJ , Maule, AG & Spithill, TW 2020, 'Molecular characterisation and vaccine efficacy of two novel developmentally regulated surface tegument proteins of Fasciola hepatica', Veterinary Parasitology, vol. 286, 109244. https://doi.org/10.1016/j.vetpar.2020.109244

TY - JOUR

T1 - Molecular characterisation and vaccine efficacy of two novel developmentally regulated surface tegument proteins of Fasciola hepatica

AU - McCusker, Paul

AU - Toet, Hayley

AU - Rathinasamy, Vignesh

AU - Young, Neil

AU - Beddoe, Travis

AU - Anderson, Glenn

AU - Dempster, Robert

AU - McVeigh, Paul

AU - McCammick, Erin

AU - Wells, Duncan

AU - Mousley, Angela

AU - Marks, Nikki J.

AU - Maule, Aaron G.

AU - Spithill, Terry W.

N1 - Funding Information: The work in the T.W.S. laboratory was supported by funds from Linkage Project LP130100943 ?Development of a novel vaccine targeting parasite tegument proteins for liver fluke disease in livestock? and Linkage Project LP160100442 ?Optimisation of a novel hybrid vaccine for liver fluke disease in cattle? funded by the Australian Research Council, in collaboration with Virbac (Australia) Pty. Ltd. In addition, support was obtained from La Trobe University, Melbourne, Australia. V.R. was supported by a Victorian-India Doctoral Scholarship from the Australia India Institute, a PhD scholarship from La Trobe University and a La Trobe University School of Life Sciences Postgraduate Publication Award. T.B. was supported by Pfizer Australia Research Fellowship. Funding from the National Health Medical Research Council (NHMRC) of Australia is gratefully acknowledged (N.D.Y.). Work at the A.G.M. lab in QUB was supported by the Professor John Glover Memorial Fund awarded to P.M. and the National Centre for the 3Rs grant NC/N001486/1. PM received an Australian Bicentennial Scholarship. We thank Prof Mike Stear for statistical analysis and Dr Wynne Gibbison (Virbac Australia) for oversight and management of the cattle vaccine trial. Funding Information: The work in the T.W.S. laboratory was supported by funds from Linkage Project LP130100943 “Development of a novel vaccine targeting parasite tegument proteins for liver fluke disease in livestock” and Linkage Project LP160100442 “Optimisation of a novel hybrid vaccine for liver fluke disease in cattle” funded by the Australian Research Council, in collaboration with Virbac (Australia) Pty. Ltd. . In addition, support was obtained from La Trobe University, Melbourne, Australia. V.R. was supported by a Victorian-India Doctoral Scholarship from the Australia India Institute , a PhD scholarship from La Trobe University and a La Trobe University School of Life Sciences Postgraduate Publication Award . T.B. was supported by Pfizer Australia Research Fellowship . Funding from the National Health Medical Research Council (NHMRC) of Australia is gratefully acknowledged (N.D.Y.). Work at the A.G.M. lab in QUB was supported by the Professor John Glover Memorial Fund awarded to P.M. and the National Centre for the 3Rs grant NC/N001486/1 . PM received an Australian Bicentennial Scholarship . We thank Prof Mike Stear for statistical analysis and Dr Wynne Gibbison (Virbac Australia) for oversight and management of the cattle vaccine trial. Publisher Copyright: © 2020 Elsevier B.V. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.

PY - 2020/10

Y1 - 2020/10

N2 - The surface tegument of Fasciola hepatica is a crucial tissue due to its key role at the host-parasite interface. We characterised three novel proteins, termed Fhteg1, Fhteg5 and Fhteg8, that are found in the tegument membrane fraction of adult F. hepatica. Bioinformatic analysis of proteomic datasets identified Fhteg5 and Fhteg8 as tegumental glycoproteins and revealed that Fhteg1, Fhteg5 and Fhteg8 are associated with exosomes of adult F. hepatica. Fhteg1, Fhteg5 and Fhteg8 appear to be related to uncharacterised sequences in F. gigantica, Fasciolopsis buski, Echinostoma caproni, Clonorchis sinensis, Opisthorchis viverrini, Schistosoma japonicum and S. mansoni, although F. hepatica appears to have expanded this family. Fhteg1 and Fhteg5 were characterised in detail. The Fhteg1 and Fhteg5 gene transcripts each demonstrate significant upregulation in juvenile fluke 2–4 days post-excystment, with transcript levels maintained during development over 3 weeks in vitro. RNAseq data showed that both Fhtegs are expressed in the adult life stage, although the transcript levels were about 8 fold lower than those in juveniles (3 week post infection). Using immunocytochemistry, Fhteg1 and Fhteg5 were each shown to be expressed in cells adjacent to the muscle layer as well as on the surface of 1 week old juveniles, whilst Fhteg5 was also present in cells at the base of the pharynx. RNAi mediated knockdown of Fhteg1 and Fhteg5 transcripts in 4–10 day old juveniles had no effect on parasite survival, movement or growth in vitro. Although no IgG responses were observed for Fhteg1 or Fhteg5 during infection in sheep and cattle, both proteins elicited a low IgG response in a proportion of infected rats. Rats vaccinated with Fhteg1 and Fhteg5 showed good IgG responses to both proteins and a mean 48.2 % reduction in worm burden following parasite challenge. Although vaccination of cattle with both proteins induced a range of IgG responses, no protection was observed against parasite challenge. This is the first study to provide insights into the molecular properties of two novel, developmentally regulated surface tegument proteins in F. hepatica.

AB - The surface tegument of Fasciola hepatica is a crucial tissue due to its key role at the host-parasite interface. We characterised three novel proteins, termed Fhteg1, Fhteg5 and Fhteg8, that are found in the tegument membrane fraction of adult F. hepatica. Bioinformatic analysis of proteomic datasets identified Fhteg5 and Fhteg8 as tegumental glycoproteins and revealed that Fhteg1, Fhteg5 and Fhteg8 are associated with exosomes of adult F. hepatica. Fhteg1, Fhteg5 and Fhteg8 appear to be related to uncharacterised sequences in F. gigantica, Fasciolopsis buski, Echinostoma caproni, Clonorchis sinensis, Opisthorchis viverrini, Schistosoma japonicum and S. mansoni, although F. hepatica appears to have expanded this family. Fhteg1 and Fhteg5 were characterised in detail. The Fhteg1 and Fhteg5 gene transcripts each demonstrate significant upregulation in juvenile fluke 2–4 days post-excystment, with transcript levels maintained during development over 3 weeks in vitro. RNAseq data showed that both Fhtegs are expressed in the adult life stage, although the transcript levels were about 8 fold lower than those in juveniles (3 week post infection). Using immunocytochemistry, Fhteg1 and Fhteg5 were each shown to be expressed in cells adjacent to the muscle layer as well as on the surface of 1 week old juveniles, whilst Fhteg5 was also present in cells at the base of the pharynx. RNAi mediated knockdown of Fhteg1 and Fhteg5 transcripts in 4–10 day old juveniles had no effect on parasite survival, movement or growth in vitro. Although no IgG responses were observed for Fhteg1 or Fhteg5 during infection in sheep and cattle, both proteins elicited a low IgG response in a proportion of infected rats. Rats vaccinated with Fhteg1 and Fhteg5 showed good IgG responses to both proteins and a mean 48.2 % reduction in worm burden following parasite challenge. Although vaccination of cattle with both proteins induced a range of IgG responses, no protection was observed against parasite challenge. This is the first study to provide insights into the molecular properties of two novel, developmentally regulated surface tegument proteins in F. hepatica.

KW - Cattle

KW - Exosome

KW - Fasciola hepatica

KW - Juvenile fluke

KW - qPCR

KW - Rat

KW - RNAi

KW - Tegument membrane proteins

KW - Vaccine

U2 - 10.1016/j.vetpar.2020.109244

DO - 10.1016/j.vetpar.2020.109244

M3 - Article

C2 - 32971381

AN - SCOPUS:85091353535

VL - 286

JO - Veterinary Parasitology

JF - Veterinary Parasitology

SN - 0304-4017

M1 - 109244

ER -

Molecular characterisation and vaccine efficacy of two novel developmentally regulated surface tegument proteins of Fasciola hepatica

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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Cite this