Abstract
Avian trichomonosis is a worldwide and cross-species epidemic, and the infection in pigeons is particularly severe. Although the disease causes a serious threat to poultry health resulting in significant economic losses, the relationship between Trichomonas gallinae (T. gallinae) and host innate immunity is still not clear. Extracellular traps (ETs) are an innate immunity response to parasitic infections. However, whether host cells can produce ETs after T. gallinae infection has not yet been reported. In the present study, the ability of T. gallinae to induce the production of heterophil extracellular traps (HETs) in pigeons was examined. T. gallinae-induced HETs were observed by scanning electron microscopy (SEM) and the main components of HETs were detected by fluorescence confocal microscopy. Changes in reactive oxygen species (ROS) and lactate dehydrogenase (LDH) were tested during the HETosis. A quantitative analysis of T. gallinae-induced HETs, the role of myeloperoxidase (MPO), store-operated Ca (2+) entry (SOCE), and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in T. gallinae-induced HET formation were conducted by inhibitor assays. The results showed that T. gallinae induced ET formation in pigeon heterophils. ETs consisted of a DNA skeleton, neutrophil elastase (NE), MPO, and Histone3 (H3). T. gallinae-induced HETs formation in a dose- and time-dependent process. The release of T. gallinae-induced HETs depends on MPO, SOCE, and NADPH oxidase. Furthermore, after T. gallinae stimulated pigeon heterophils, ROS production was significantly increased, while no significant differences in the LDH activity were observed.
Graphical Abstract
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The datasets generated during the current study are available from the corresponding author on reasonable request.
References
Abi Abdallah DS, Lin C, Ball CJ, King MR, Duhamel GE, Denkers EY (2012) Toxoplasma gondii triggers release of human and mouse neutrophil extracellular traps. Infect Immun 80:768–777. https://doi.org/10.1128/IAI.05730-11
Adinehbeigi K, Ebrahimi M, SoltaniEini M, Sameie A (2018) Prevalence of Haemoproteus columbae (Apicomplexa: Haemoproteidae) and Trichomonas gallinae (Metamonada: Trichomonadidae) infections among pigeons (Columba livia) in West Azerbaijan Province Iran. Arch Razi Inst 73:147–152. https://doi.org/10.22092/ari.2018.116619
Amin A, Bilic I, Liebhart D, Hess M (2014) Trichomonads in birds–a review. Parasitology 141:733–747. https://doi.org/10.1017/S0031182013002096
Andrieux P, Chevillard C, Cunha-Neto E, Nunes JPS (2021) Mitochondria as a cellular hub in infection and inflammation. Int J Mol Sci 22(21):11338. https://doi.org/10.3390/ijms222111338
Bennike TB, Carlsen TG, Ellingsen T, Bonderup OK, Glerup H, Bøgsted M, Christiansen G, Birkelund S, Stensballe A, Andersen V (2015) Neutrophil extracellular traps in Ulcerative Colitis: a proteome analysis of intestinal biopsies. Inflamm Bowel Dis 21:2052–2067. https://doi.org/10.1097/MIB.0000000000000460
Borji H, Razmi GH, Movassaghi AH, Moghaddas E, Azad M (2011) Prevalence and pathological lesion of Trichomonas gallinae in pigeons of Iran. J Parasit Dis 35:186–189. https://doi.org/10.1007/s12639-011-0047-2
Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, Weinrauch Y, Zychlinsky A (2004) Neutrophil extracellular traps kill bacteria. Science 303:1532–1535. https://doi.org/10.1126/science.1092385
Chhabra R, Chantrey J, Ganapathy K (2015) Immune responses to virulent and vaccine strains of infectious bronchitis viruses in chickens. Viral Immunol. 28(9):478–488. https://doi.org/10.1089/vim.2015.0027
Chi JF, Lawson B, Durrant C, Beckmann K, John S, Alrefaei AF, Kirkbride K, Bell DJ, Cunningham AA, Tyler KM (2013) The finch epidemic strain of Trichomonas gallinae is predominant in British non-passerines. Parasitology 140:1234–1245. https://doi.org/10.1017/S0031182013000930
Eruslanov E, Kusmartsev S (2010) Identification of ROS using oxidized DCFDA and flow-cytometry. Methods Mol Biol 594:57–72. https://doi.org/10.1007/978-1-60761-411-1_4
Genovese KJ, He H, Swaggerty CL, Kogut MH (2013) The avian heterophil. Dev Comp Immunol 41:334–340. https://doi.org/10.1007/978-1-60761-411-1_4
Gerhold RW, Yabsley MJ, Smith AJ, Ostergaard E, Mannan W, Cann JD, Fischer JR (2008) Molecular characterization of the Trichomonas gallinae morphologic complex in the United States. J Parasitol 94:1335–1341. https://doi.org/10.1645/GE-1585.1
Grob D, Conejeros I, Velásquez ZD, Preußer C, Gärtner U, Alarcón P, Burgos RA, Hermosilla C, Taubert A (2020) Trypanosoma brucei induces polymorphonuclear neutrophil activation and neutrophil extracellular traps release. Front Immunol 11:559561. https://doi.org/10.3389/fimmu.2020.559561
Grunenwald C, Sidor I, Mickley R, Dwyer C, Gerhold R (2018) Tetratrichomonas and Trichomonas spp.-associated disease in free-ranging common eiders (Somateria mollissima) from Wellfleet Bay, MA and description of ITS1 region genotypes. Avian Dis 62:117–123. https://doi.org/10.1637/11742-080817-Reg.1
Guo Q, Zhu Z, Wang J, Huang W, Zhang C, Zeng J, Zhao H, Qi T, Zhou W, Zhang T, Zhang C, Xiao F (2021) Preparation, stability and commutability of candidate reference materials for lactate dehydrogenase (LDH). Clin Biochem 91:45–51. https://doi.org/10.1016/j.clinbiochem.2021.02.002
Hemmers S, Teijaro JR, Arandjelovic S, Mowen KA (2011) PAD4-mediated neutrophil extracellular trap formation is not required for immunity against influenza infection. PLoS One 6:e22043. https://doi.org/10.1111/jth.15313
Jiang X, Sun J, Wang F, Li H, Zhao X (2016) Prevalence of Trichomonas spp. in domestic pigeons in Shandong Province, China, and genotyping by restriction fragment length polymorphism. Vet J 211:88–93. https://doi.org/10.1016/j.tvjl.2015.10.029
King PT, Sharma R, O’Sullivan K, Selemidis S, Lim S, Radhakrishna N, Lo C, Prasad J, Callaghan J, McLaughlin P, Farmer M, Steinfort D, Jennings B, Ngui J, Broughton BR, Thomas B, Essilfie AT, Hickey M, Holmes PW, Hansbro P, Bardin PG, Holdsworth SR (2015) Nontypeable Haemophilus influenzae induces sustained lung oxidative stress and protease expression. PLoS One 10:e0120371. https://doi.org/10.1371/journal.pone.0120371
Knackstedt SL, Georgiadou A, Apel F, Abu-Abed U, Moxon CA, Cunnington AJ, Raupach B, Cunningham D, Langhorne J, Krüger R, Barrera V, Harding SP, Berg A, Patel S, Otterdal K, Mordmüller B, Schwarzer E, Brinkmann V, Zychlinsky A, Amulic B (2019) Neutrophil extracellular traps drive inflammatory pathogenesis in malaria. Sci Immunol 4(40):eaaw0336. https://doi.org/10.1126/sciimmunol.aaw0336
Li L, Li X, Li G, Gong P, Zhang X, Yang Z, Yang J, Li J (2018) Mouse macrophages capture and kill Giardia lamblia by means of releasing extracellular trap. Dev Comp Immunol 88:206–212. https://doi.org/10.1016/j.dci.2018.07.024
Marx M, Reiner G, Willems H, Rocha G, Hillerich K, Masello JF, Mayr SL, Moussa S, Dunn JC, Thomas RC, Goodman SJ, Hamer KC, Metzger B, Cecere JG, Spina F, Koschkar S, Calderón L, Romeike T, Quillfeldt P (2017) High prevalence of Trichomonas gallinae in wild columbids across western and southern Europe. Parasit Vectors 10:242. https://doi.org/10.1186/s13071-017-2170-0
Matoszka N, Działo J, Tokarz-Deptuła B, Deptuła W (2012) NET and NETosis–new phenomenon in immunology. Postepy Hig Med Dosw 66:437–445. https://doi.org/10.5604/17322693.1001178
McBurney S, Kelly-Clark WK, Forzán MJ, Lawson B, Tyler KM, Greenwood SJ (2015) Molecular characterization of Trichomonas gallinae isolates recovered from the Canadian Maritime provinces’ wild avifauna reveals the presence of the genotype responsible for the European finch trichomonosis epidemic and additional strains. Parasitology 142(8):1053–1062. https://doi.org/10.1017/S0031182015000281
Merling de Chapa M, Auls S, Kenntner N, Krone O (2021) To get sick or not to get sick—Trichomonas infections in two Accipiter species from Germany. Parasitol Res 120:3555–3567. https://doi.org/10.1007/s00436-021-07299-1
Muñoz-Caro T, Lendner M, Daugschies A, Hermosilla C, Taubert A (2015) NADPH oxidase, MPO, NE, ERK1/2, p38 MAPK and Ca2+ influx are essential for Cryptosporidium parvum-induced NET formation. Dev Comp Immunol 52:245–254. https://doi.org/10.1016/j.dci.2015.05.007
Muñoz-Caro T, Mena Huertas SJ, Conejeros I, Alarcón P, Hidalgo MA, Burgos RA, Hermosilla C, Taubert A (2015) Eimeria bovis-triggered neutrophil extracellular trap formation is CD11b-, ERK 1/2-, p38 MAP kinase- and SOCE-dependent. Vet Res 46:23. https://doi.org/10.1186/s13567-015-0155-6
Muñoz-Caro T, Silva LM, Ritter C, Taubert A, Hermosilla C (2014) Besnoitia besnoiti tachyzoites induce monocyte extracellular trap formation. Parasitol Res 113:4189–4197. https://doi.org/10.1007/s00436-014-4094-3
Parhamifar L, Andersen H, Moghimi SM (2019) Lactate Dehydrogenase Assay for Assessment of Polycation Cytotoxicity. Methods Mol Biol 1943:291–299. https://doi.org/10.1007/978-1-4939-9092-4_18
Pilsczek FH, Salina D, Poon KK, Fahey C, Yipp BG, Sibley CD, Robbins SM, Green FH, Surette MG, Sugai M, Bowden MG, Hussain M, Zhang K, Kubes P (2010) A novel mechanism of rapid nuclear neutrophil extracellular trap formation in response to Staphylococcus aureus. J Immunol 185:7413–7425. https://doi.org/10.4049/jimmunol.1000675
Reichel M, Muñoz-Caro T, Sanchez Contreras G, Rubio García A, Magdowski G, Gärtner U, Taubert A, Hermosilla C (2015) Harbour seal (Phoca vitulina) PMN and monocytes release extracellular traps to capture the apicomplexan parasite Toxoplasma gondii. Dev Comp Immunol 50:106–115. https://doi.org/10.1016/j.dci.2015.02.002
Rochael NC, Guimarães-Costa AB, Nascimento MT, DeSouza-Vieira TS, Oliveira MP, Garcia e Souza LF, Oliveira MF, Saraiva EM (2015) Classical ROS-dependent and early/rapid ROS-independent release of neutrophil extracellular traps triggered by Leishmania parasites. Sci Rep 5:18302. https://doi.org/10.1038/srep18302
Saitoh T, Komano J, Saitoh Y, Misawa T, Takahama M, Kozaki T, Uehata T, Iwasaki H, Omori H, Yamaoka S, Yamamoto N, Akira S (2012) Neutrophil extracellular traps mediate a host defense response to human immunodeficiency virus-1. Cell Host Microbe 12:109–116. https://doi.org/10.1016/j.chom.2012.05.015
Scherz-Shouval R, Elazar Z (2011) Regulation of autophagy by ROS: physiology and pathology. Trends Biochem Sci 36:30–38. https://doi.org/10.1016/j.tibs.2010.07.007
Silva LM, Caro TM, Gerstberger R, Vila-Viçosa MJ, Cortes HC, Hermosilla C, Taubert A (2014) The apicomplexan parasite Eimeria arloingi induces caprine neutrophil extracellular traps. Parasitol Res 113:2797–2807. https://doi.org/10.1007/s00436-014-3939-0
Stoiber W, Obermayer A, Steinbacher P, Krautgartner WD (2015) The Role of reactive oxygen species (ROS) in the formation of extracellular traps (ETs) in humans. Biomolecules 5:702–723. https://doi.org/10.3390/biom5020702
Wei R, Li X, Wang X, Wang Y, Zhang X, Zhang N, Wang J, Yang J, Zhang X, Gong P, Li J (2021) Trypanosoma evansi triggered neutrophil extracellular traps formation dependent on myeloperoxidase, neutrophil elastase, and extracellular signal-regulated kinase 1/2 signaling pathways. Vet Parasitol 296:109502. https://doi.org/10.1016/j.vetpar.2021.109502
Wei Z, Hermosilla C, Taubert A, He X, Wang X, Gong P, Li J, Yang Z, Zhang X (2016) Canine neutrophil extracellular traps release induced by the apicomplexan parasite Neospora caninum in vitro. Front Immunol 7:436. https://doi.org/10.3389/fimmu.2016.00436
Wei Z, Wang Y, Zhang X, Wang X, Gong P, Li J, Taubert A, Hermosilla C, Zhang X, Yang Z (2018) Bovine macrophage-derived extracellular traps act as early effectors against the abortive parasite Neospora caninum. Veterinary parasitology 258:1–7. https://doi.org/10.1016/j.vetpar.2018.06.002
Wei Z, Zhao Y, Zhang N, Han Z, Liu X, Jiang A, Zhang Y, Wang C, Gong P, Li J, Zhang X, Yang Z (2019) Eimeria tenella induces the release of chicken heterophil extracellular traps. Veterinary parasitology 275:108931. https://doi.org/10.1016/j.vetpar.2019.108931
Yang Z, Wei Z, Hermosilla C, Taubert A, He X, Wang X, Gong P, Li J, Zhang X (2017) Caprine Monocytes Release Extracellular Traps against Neospora caninum In Vitro. Front Immunol 8:2016. https://doi.org/10.3389/fimmu.2017.02016
Acknowledgements
We thank Chengyao Li, Cong Ding, Mengge Chen, Heng Yang, and Xiaodan Yuan for the excellent laboratory work and technical support. We thank Li Pan and Yi Xin for helping us with the SEM analysis. We thank Xinrui Wang for the use of the Olympus FluoView FV1000 confocal microscope.
Funding
We thank the support from key projects of the national key R & D plan and intergovernmental international scientific and technological innovation cooperation (2022YFE0114500) .
Author information
Authors and Affiliations
Contributions
Jianhua Li, Yuru Wang, and Xin Li supervised the study and design. Hongyu Wang, Xuehan Wang, Ran Wei, and Nan Zhang conducted these experiments and wrote the manuscript. Xiaocen Wang, Xichen Zhang, and Pengtao Gong revised the manuscript. All authors have reviewed the manuscript.
Corresponding authors
Ethics declarations
Ethics approval
The authors confirm the ethical policies of the journal and complied with these policies in the experiment. These experiments were conducted as approved by the Ethics Committee of Jilin University on the Care and Use of Laboratory Animals (Project number: JLU20170330).
Consent to participate
Not applicable.
Consent for publication
Not applicable.
Conflict of interest
The authors declare no competing interests.
Additional information
Section Editor: Yaoyu Feng
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 (e.g. a society or other partner) 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
Wang, H., Wang, Y., Wang, X. et al. Trichomonas gallinae induces heterophil extracellular trap formation in pigeons. Parasitol Res 122, 527–536 (2023). https://doi.org/10.1007/s00436-022-07755-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-022-07755-6