Abstract
Liver flukes, Fasciola spp., are veterinary and medically important parasites infecting numerous species of economically important animals in addition to humans on a global scale. The components of transforming growth factor beta (TGF-β) signalling are widely distributed throughout the animal kingdom and are considerably conserved. Through shared common signal transduction mechanisms, crosstalk of TGF-β signalling between a host and the parasite during infection is possible. Herein, we have identified and undertaken the molecular characterisation of a putative TGF-β homologue from the tropical liver fluke F. gigantica (FgTLM). A FgTLM cDNA was 3557 bp in length, it encoded for 620 amino acid polypeptide which consisted of 494 amino acids of prodomain and 126 amino acids comprising the mature protein. FgTLM displayed characteristic structures of mammalian TGF-β ligands that were unique to the inhibin-β chain, monomer of activin. A phylogenetic analysis revealed the high degree of conservation with TGF-β molecules from trematode species. Interestingly, the sequence of amino acid in the active domain of FgTLM was completely identical to FhTLM from F. hepatica. FgTLM expressed throughout the lifecycle of F. gigantica but was highly expressed in developmental active stages. The dynamics of expression of FgTLM during the developmental stages of F. gigantica was comparable to the pattern of TGF-β expression in F. hepatica. Our findings demonstrated that FgTLM exhibits a high level of similarity to FhTLM in the context of both amino acid sequence and the life stage expression patterns. These similarities underline the possibility that the FgTLM molecule might have the same properties and functions as FhTLM in biological processes of the immature parasites and host immune evasion. Consequently, the specific biological functions of FgTLM on either parasite or relevant hosts need to be defined experimentally.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data generated or analysed during this study are included in this published article.
References
Adalid-Peralta L, Rosas G, Arce-Sillas A, Bobes RJ (2017) Effect of transforming growth factor-β upon Taenia solium and Taenia crassiceps cysticerci. Sci Rep 7(1):12345. https://doi.org/10.1038/s41598-017-12202-z
Batlle E, Massagué J (2019) Transforming growth factor-β signaling in immunity and cancer. Immunity 50(4):924–940. https://doi.org/10.1016/j.immuni.2019.03.024
Beesley NJ, Caminade C, Charlier J, Flynn RJ, Hodgkinson JE, Martinez-Moreno A, Martinez-Valladares M, Perez J, Rinaldi L, Williams DJ (2018) Fasciola and fasciolosis in ruminants in Europe: identifying research needs. Transbound Emerg Dis 65(S1):199–216. https://doi.org/10.1111/tbed.12682
Donnelly S, Flynn R, Mulcahy G, O’Neill S (2021) Immunological interaction between Fasciola and its host Fasciolosis. CABI, Wallingford, pp 278–307
Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32(5):1792–1797. https://doi.org/10.1093/nar/gkh340
Flynn RJ, Mulcahy G, Elsheikha HM (2010) Coordinating innate and adaptive immunity in Fasciola hepatica infection: implications for control. Vet Parasitol 169(3):235–240. https://doi.org/10.1016/j.vetpar.2010.02.015
Flynn RJ, Mulcahy G, Welsh M, Cassidy JP, Corbett D, Milligan C, Andersen P, Strain S, McNair J (2009) Co-infection of cattle with Fasciola hepatica and Mycobacterium bovis- immunological consequences. Transbound Emerg Dis 56(6–7):269–274. https://doi.org/10.1111/j.1865-1682.2009.01075.x
Freitas TC, Jung E, Pearce EJ (2007) TGF-β signaling controls embryo development in the parasitic flatworm Schistosoma mansoni. PLoS Pathog 3(4):e52. https://doi.org/10.1371/journal.ppat.0030052
Gomez-Escobar N, Gregory WF, Maizels RM (2000) Identification of tgh-2, a filarial nematode homolog of Caenorhabditis elegans daf-7 and human transforming growth factor beta, expressed in microfilarial and adult stages of Brugia malayi. Infect Immun 68(11):6402–6410. https://doi.org/10.1128/IAI.68.11.6402-6410.2000
Gouy M, Guindon S, Gascuel O (2009) SeaView Version 4: a multiplatform graphical user interface for sequence alignment and phylogenetic tree building. Mol Biol Evol 27(2):221–224. https://doi.org/10.1093/molbev/msp259
Grainger JR, Smith KA, Hewitson JP, McSorley HJ, Harcus Y, Filbey KJ, Finney CA, Greenwood EJ, Knox DP, Wilson MS, Belkaid Y (2010) Helminth secretions induce de novo T cell Foxp3 expression and regulatory function through the TGF-β pathway. J Exp Med 207(11):2331–2341. https://doi.org/10.1084/jem.20101074
Hall T (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98
He L, Liu H, Zhang BY, Li FF, Di WD, Wang CQ, Zhou CX, Liu L, Li TT, Zhang T, Fang R (2020) A daf-7-related TGF-β ligand (Hc-tgh-2) shows important regulations on the development of Haemonchus contortus. Parasit Vectors 13(1):326. https://doi.org/10.1186/s13071-020-04196-x
Helmby H (2015) Human helminth therapy to treat inflammatory disorders - where do we stand? BMC Immunol 16:12. https://doi.org/10.1186/s12865-015-0074-3
Hewitson JP, Grainger JR, Maizels RM (2009) Helminth immunoregulation: the role of parasite secreted proteins in modulating host immunity. Mol Biochem Parasitol 167(1):1–11. https://doi.org/10.1016/j.molbiopara.2009.04.008
Hinck AP, Mueller TD, Springer TA (2016) Structural biology and evolution of the TGF-β family. Cold Spring Harb Perspect Biol 8(12). https://doi.org/10.1101/cshperspect.a022103
Howe KL, Bolt BJ, Cain S, Chan J, Chen WJ, Davis P, Done J, Down T, Gao S, Grove C, Harris TW (2015) WormBase 2016: expanding to enable helminth genomic research. Nucleic Acids Res 44(D1):D774–D780. https://doi.org/10.1093/nar/gkv1217
Howe KL, Bolt BJ, Shafie M, Kersey P, Berriman M (2017) WormBase ParaSite − a comprehensive resource for helminth genomics. Mol Biochem Parasitol 215:2–10. https://doi.org/10.1016/j.molbiopara.2016.11.005
Japa O, Hodgkinson JE, Emes RD, Flynn RJ (2015) TGF-β superfamily members from the helminth Fasciola hepatica show intrinsic effects on viability and development. Vet Res 46:29. https://doi.org/10.1186/s13567-015-0167-2
Johnston CJC, Smyth DJ, Kodali RB, White MP, Harcus Y, Filbey KJ, Hewitson JP, Hinck CS, Ivens A, Kemter AM, Kildemoes AO (2017) A structurally distinct TGF-β mimic from an intestinal helminth parasite potently induces regulatory T cells. Nat Commun 8(1):1741. https://doi.org/10.1038/s41467-017-01886-6
Kelley LA, Mezulis S, Yates CM, Wass MN, Sternberg MJE (2015) The Phyre2 web portal for protein modeling, prediction and analysis. Nat Protoc 10(6):845–858. https://doi.org/10.1038/nprot.2015.053
Kinjo AR, Nishikawa K (2004) Eigenvalue analysis of amino acid substitution matrices reveals a sharp transition of the mode of sequence conservation in proteins. Bioinformatics 20(16):2504–2508. https://doi.org/10.1093/bioinformatics/bth297
Lalor R, Cwiklinski K, Calvani NE, Dorey A, Hamon S, Corrales JL, Dalton JP, De Marco VC (2021) Pathogenicity and virulence of the liver flukes Fasciola hepatica and Fasciola gigantica that cause the zoonosis Fasciolosis. Virulence 12(1):2839–2867. https://doi.org/10.1080/21505594.2021.1996520
Maizels RM, Smits HH, McSorley HJ (2018) Modulation of host immunity by helminths: the expanding repertoire of parasite effector molecules. Immunity 49(5):801–818. https://doi.org/10.1016/j.immuni.2018.10.016
Massagué J, Chen YG (2000) Controlling TGF-β signaling. Genes Dev 14(6):627–644. https://doi.org/10.1101/gad.14.6.627
McVeigh P, McCammick EM, McCusker P, Morphew RM, Mousley A, Abidi A, Saifullah KM, Muthusamy R, Gopalakrishnan R, Spithill TW, Dalton JP (2014) RNAi dynamics in juvenile Fasciola spp. Liver flukes reveals the persistence of gene silencing in vitro. PLOS Negl Trop Dis 8(9):e3185. https://doi.org/10.1371/journal.pntd.0003185
Moxon JV, LaCourse EJ, Wright HA, Perally S, Prescott MC, Gillard JL, Barrett J, Hamilton JV, Brophy PM (2010) Proteomic analysis of embryonic Fasciola hepatica: characterization and antigenic potential of a developmentally regulated heat shock protein. Vet Parasitol 169(1):62–75. https://doi.org/10.1016/j.vetpar.2009.12.031
Musah-Eroje M, Hoyle RC, Japa O, Hodgkinson JE, Haig DM, Flynn RJ (2021) A host-independent role for Fasciola hepatica transforming growth factor-like molecule in parasite development. Int J Parasitol 51(6):481–492. https://doi.org/10.1016/j.ijpara.2020.11.005
Nono JK, Lutz MB, Brehm K (2019) A secreted Echinococcus multilocularis activin A homologue promotes regulatory T cell expansion. BioRxiv:618140. https://doi.org/10.1101/618140
Nyirenda SS, Sakala M, Moonde L, Kayesa E, Fandamu P, Banda F, Sinkala Y (2019) Prevalence of bovine fascioliasis and economic impact associated with liver condemnation in abattoirs in Mongu district of Zambia. BMC Vet Res 15(1):33–33. https://doi.org/10.1186/s12917-019-1777-0
Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, Ferrin TE (2004) UCSF Chimera–a visualization system for exploratory research and analysis. J Comput Chem 25(13):1605–1612. https://doi.org/10.1002/jcc.20084
Piedrafita D, Spithill TW, Smith RE, Raadsma HW (2010) Improving animal and human health through understanding liver fluke immunology. Parasite Immunol 32(8):572–581. https://doi.org/10.1111/j.1365-3024.2010.01223.x
Richards GS, Degnan BM (2009) The dawn of developmental signaling in the metazoa. Cold Spring Harb Symp Quant Biol 74:81–90. https://doi.org/10.1101/sqb.2009.74.028
Robinson MW, Dalton JP (2009) Zoonotic helminth infections with particular emphasis on fasciolosis and other trematodiases. Philos Trans R Soc B Biol Sci 364(1530):2763–2776. https://doi.org/10.1098/rstb.2009.0089
Rost B (1999) Twilight zone of protein sequence alignments. Protein Eng 12(2):85–94. https://doi.org/10.1093/protein/12.2.85
Ryan S, Shiels J, Taggart CC, Dalton JP, Weldon S (2020) Fasciola hepatica-derived molecules as regulators of the host immune response. 11. https://doi.org/10.3389/fimmu.2020.02182
Sargison ND, Scott PR (2011) Diagnosis and economic consequences of triclabendazole resistance in Fasciola hepatica in a sheep flock in south-east Scotland. Vet Rec 168(6):159. https://doi.org/10.1136/vr.c5332
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9(7):671–675. https://doi.org/10.1038/nmeth.2089
Sulaiman AA, Zolnierczyk K, Japa O, Owen JP, Maddison BC, Emes RD, Hodgkinson JE, Gough KC, Flynn RJ (2016) A trematode parasite derived growth factor binds and exerts influences on host immune functions via host cytokine receptor complexes. PLoS Pathogen 12(11):e1005991. https://doi.org/10.1371/journal.ppat.1005991
Teufel F, Almagro Armenteros JJ, Johansen AR, Gíslason MH, Pihl SI, Tsirigos KD, Winther O, Brunak S, von Heijne G, Nielsen H (2022) SignalP 6.0 predicts all five types of signal peptides using protein language models. Nat Biotechnol. https://doi.org/10.1038/s41587-021-01156-3
van der Zande HJP, Zawistowska-Deniziak A, Guigas B (2019) Immune regulation of metabolic homeostasis by helminths and their molecules. Trends Parasitol 35(10):795–808. https://doi.org/10.1016/j.pt.2019.07.014
Whelan S, Goldman N (2001) A general empirical model of protein evolution derived from multiple protein families using a maximum-likelihood approach. Mol Biol Evol 18(5):691–699. https://doi.org/10.1093/oxfordjournals.molbev.a003851
Wu MY, Hill CS (2009) TGF-β superfamily signaling in embryonic development and homeostasis. Dev Cell 16(3):329–343. https://doi.org/10.1016/j.devcel.2009.02.012
Zhang XX, Cwiklinski K, Hu RS, Zheng WB, Sheng ZA, Zhang FK, Elsheikha HM, Dalton JP, Zhu XQ (2019) Complex and dynamic transcriptional changes allow the helminth Fasciola gigantica to adjust to its intermediate snail and definitive mammalian hosts. BMC Genomics 20(1):729–729. https://doi.org/10.1186/s12864-019-6103-5
Acknowledgements
The authors are grateful to technical assistance provided by Ms. Chorpaka Puangsri.
Funding
This research project was supported by the Thailand Science Research and Innovation Fund and the University of Phayao (grant no. FF64-RIB007), Phayao, Thailand.
Author information
Authors and Affiliations
Contributions
Ornampai Japa and Robin J Flynn conceived and designed the study. Ornampai Japa collected and maintained the parasites. Ornampai Japa performed the experiments, data collection and bioinformatics analyses. Ornampai Japa, Khanuengnij Prakhammin and Robin J Flynn analysed and interpreted the data. Ornampai Japa and Robin J Flynn drafted and revised the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Ethics approval
Animal ethics and study protocol approval were granted by the Animal Ethics Committee of the University of Phayao, Thailand, approval number 63 01 04 006.
Consent to participate
All authors confirm their participation in the study.
Consent for publication
All authors consent to publication of study data.
Competing interests
The authors declare no competing interests.
Additional information
Handling Editor: Julia Walochnik
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Japa, O., Prakhammin, K. & Flynn, R.J. Identification and expression of a transforming growth factor beta (TGF-β) homologue in the tropical liver fluke Fasciola gigantica. Parasitol Res 121, 3547–3559 (2022). https://doi.org/10.1007/s00436-022-07679-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-022-07679-1