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The effect of iron on Trichomonas vaginalis TvCP2: a cysteine proteinase found in vaginal secretions of trichomoniasis patients

Published online by Cambridge University Press:  16 March 2020

Luis Alberto Rivera-Rivas
Affiliation:
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), CP 07360, Mexico City, Mexico
Sebastián Lorenzo-Benito
Affiliation:
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), CP 07360, Mexico City, Mexico
Diana Belén Sánchez-Rodríguez
Affiliation:
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), CP 07360, Mexico City, Mexico
Jesús FT Miranda-Ozuna
Affiliation:
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), CP 07360, Mexico City, Mexico
Esly Alejandra Euceda-Padilla
Affiliation:
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), CP 07360, Mexico City, Mexico
Jaime Ortega-López
Affiliation:
Departamento de Biotecnología y Bioingeniería, CINVESTAV-IPN, CP 07360, Mexico City, Mexico
Bibiana Chávez-Munguía
Affiliation:
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), CP 07360, Mexico City, Mexico
Anel Lagunes-Guillén
Affiliation:
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), CP 07360, Mexico City, Mexico
Beatriz Velázquez-Valassi
Affiliation:
Departamento de Vigilancia Epidemiológica del Hospital General de México ‘Eduardo Liceaga’, CP 06720, Mexico City, Mexico
Lidia Jasso-Villazul
Affiliation:
Unidad de Medicina Preventiva del Hospital General de México ‘Eduardo Liceaga’, CP 06720, Mexico City, Mexico
Rossana Arroyo*
Affiliation:
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), CP 07360, Mexico City, Mexico
*
Author for correspondence: Rossana Arroyo, E-mail: [email protected]

Abstract

Trichomonas vaginalis (Tv) induces host cell damage through cysteine proteinases (CPs) modulated by iron. An immunoproteomic analysis showed that trichomoniasis patient sera recognize various CPs, also some of them are present in vaginal washes (VWs). Thus, the goal of this work was to determine whether TvCP2 is expressed during infection and to assess the effect of iron on TvCP2 expression, localization and contribution to in vitro cellular damage. Western-blotting (WB) assays using TvCP2r and vaginitis patient serum samples showed that 6/9 Tv (+) but none of the Tv (−) patient sera recognized TvCP2r. WB using an anti-TvCP2r antibody and VWs from the same patients showed that in all of the Tv (+) but none of the Tv (−) VWs, the anti-TvCP2r antibody detected a 27 kDa protein band that corresponded to the mature TvCP2, which was confirmed by mass spectrometry analysis. Iron decreased the amount of TvCP2 mRNA and the protein localized on the parasite surface and cytoplasmic vesicles concomitant with the cytotoxic effect of TvCP2 on HeLa cells. Parasites pretreated with the anti-TvCP2r antibody also showed reduced levels of cytotoxicity and apoptosis induction in HeLa cell monolayers. In conclusion, these results show that TvCP2 is expressed during trichomonal infection and plays an important role in the in vitro HeLa cell cytotoxic damage under iron-restricted conditions.

Type
Research Article
Copyright
Copyright © The Author(s) 2020. Published by Cambridge University Press

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References

Adler, J and Parmryd, I (2010) Quantifying colocalization by correlation: the Pearson correlation coefficient is superior to the Mander's overlap coefficient. Cytometry Part A, 77, 732742. doi: 10.1002/cyto.a.20896.Google ScholarPubMed
Alderete, JF, Provenzano, D and Lehker, MW (1995) Iron mediates Trichomonas vaginalis resistance to complement lysis. Microbial Pathogens 19, 93103.10.1006/mpat.1995.0049CrossRefGoogle ScholarPubMed
Alvarez-Sánchez, ME, Ávila-González, L, Becerril-García, C, Fattel-Facenda, LV, Ortega-López, J and Arroyo, R (2000) A novel cysteine proteinase (CP65) of Trichomonas vaginalis involved in cytotoxicity. Microbial Pathogens 28, 193202.10.1006/mpat.1999.0336CrossRefGoogle ScholarPubMed
Alvarez-Sánchez, ME, Solano-González, E, Yáñez-Gómez, C and Arroyo, R (2007) Negative iron regulation of the CP65 cysteine proteinase cytotoxicity in Trichomonas vaginalis. Microbes and Infection 9, 15971605.10.1016/j.micinf.2007.09.011CrossRefGoogle ScholarPubMed
Arroyo, R and Alderete, JF (1989) Trichomonas vaginalis surface proteinase activity is necessary for adherence to epithelial cells. Infection and Immunity 57, 29912997.10.1128/IAI.57.10.2991-2997.1989CrossRefGoogle ScholarPubMed
Arroyo, R and Alderete, JF (1995) Two Trichomonas vaginalis surface proteinases bind to host epithelial cells and are related to levels of cytoadherence and cytotoxicity. Archives of Medical Research 26, 279285.Google ScholarPubMed
Arroyo, R, Cárdenas-Guerra, RE, Figueroa-Angulo, EE, Puente-Rivera, J, Zamudio-Prieto, O and Ortega-López, J (2015) Trichomonas vaginalis cysteine proteinases: iron response in gene expression and proteolytic activity. Biomed Research International 2015, 946787. doi: 10.1155/2015/946787.CrossRefGoogle ScholarPubMed
Benchimol, M (2004) Trichomonads under microscopy. Microscopy and Microanalysis 10, 528550.10.1017/S1431927604040905CrossRefGoogle ScholarPubMed
Cárdenas-Guerra, RE, Arroyo, R, Rosa de Andrade, I, Benchimol, M and Ortega-López, J (2013) The iron-induced cysteine proteinase TvCP4 plays a key role in Trichomonas vaginalis haemolysis. Microbes and Infection 15, 958968.10.1016/j.micinf.2013.09.002CrossRefGoogle Scholar
Cárdenas-Guerra, RE, Ortega-López, J, Flores-Pucheta, CI, Benítez-Cardoza, CG and Arroyo, R (2015) The recombinant prepro region of TvCP4 is an inhibitor of cathepsin L-like cysteine proteinases of Trichomonas vaginalis that inhibits trichomonal haemolysis. International Journal of Biochemistry and Cell Biology 59, 7383.10.1016/j.biocel.2014.12.001CrossRefGoogle ScholarPubMed
Carlton, JM, Hirt, RP, Silva, JC, Delcher, AL, Schatz, M, Zhao, Q, Wortman, JR, Bidwell, SL, Alsmark, UC, Besteiro, S, Sicheritz-Ponten, T, Noel, CJ, Dacks, JB, Foster, PG, Simillion, C, Van de Peer, Y, Miranda-Saavedra, D, Barton, GJ, Westrop, GD, Müller, S, Dessi, D, Fiori, PL, Ren, Q, Paulsen, I, Zhang, H, Bastida-Corcuera, FD, Simoes-Barbosa, A, Brown, MT, Hayes, RD, Mukherjee, M, Okumura, CY, Schneider, R, Smith, AJ, Vanacova, S, Villalvazo, M, Haas, BJ, Pertea, M, Feldblyum, TV, Utterback, TR, Shu, CL, Osoegawa, K, de Jong, PJ, Hrdy, I, Horvathova, L, Zubacova, Z, Dolezal, P, Malik, SB, Logsdon, JM Jr, Henze, K, Gupta, A, Wang, CC, Dunne, RL, Upcroft, JA, Upcroft, P, White, O, Salzberg, SL, Tang, P, Chiu, CH, Lee, YS, Embley, TM, Coombs, GH, Mottram, JC, Tachezy, J, Fraser-Liggett, CM and Johnson, PJ (2007) Draft genome sequence of the sexually transmitted pathogen Trichomonas vaginalis. Science (New York, N.Y.) 315, 207212.10.1126/science.1132894CrossRefGoogle ScholarPubMed
Carvajal-Gámez, BI, Quintas-Granados, LI, Arroyo, R, Vázquez-Carrillo, LI, Ramón-Luing, LL, Carrillo-Tapia, E and Álvarez-Sánchez, ME (2014) Putrescine-dependent re-localization of TvCP39, a cysteine proteinase involved in Trichomonas vaginalis cytotoxicity. PLoS ONE 9, e107293.10.1371/journal.pone.0107293CrossRefGoogle ScholarPubMed
Conrad, MD, Gorman, AW, Schillinger, JA, Fiori, PL, Arroyo, R, Malla, N, Dubey, ML, González, J, Blank, S, Secor, WE and Carlton, JM (2012) Extensive genetic diversity, unique population structure and evidence of genetic exchange in the sexually transmitted parasite Trichomonas vaginalis. PLoS Neglected Tropical Diseases 6, e1573.10.1371/journal.pntd.0001573CrossRefGoogle ScholarPubMed
Dailey, DC, Chang, TH and Alderete, JF (1990) Characterization of Trichomonas vaginalis haemolysis. Parasitology 101, 171175.10.1017/S0031182000063204CrossRefGoogle ScholarPubMed
De Jesus, JB, Ferreira, MA, Cuervo, P, Britto, C, Silva-Filho, FC and Meyer-Fernandes, JR (2006) Iron modulates ecto-phosphohydrolase activities in pathogenic trichomonads. Parasitoloy Internacional 55, 285290.Google ScholarPubMed
De Jesus, JB, Cuervo, P, Junqueira, M, Britto, C, Silva-Filho, FC, Soares, MJ, Cupolillo, E, Fernandes, O and Domont, GB (2007) A further proteomic study on the effect of iron in the human pathogen Trichomonas vaginalis. Proteomics 7, 19611972.10.1002/pmic.200600797CrossRefGoogle Scholar
Diamond, LS (1957) The establishment of various trichomonads of animals and man in axenic cultures. Journal of Parasitology 43, 488490.10.2307/3274682CrossRefGoogle ScholarPubMed
Dias-Lopes, G, Saboia-Vahia, L, Margotti, ET, Fernandes, NS, Castro, CLF, Oliveira, FO, Peixoto, JF, Britto, C, Silva-Filho, FCE, Cuervo, P and Jesus, JB (2017) Morphologic study of the effect of iron on pseudocysts formation un Trichomonas vaginalis and its interaction with human epithelial cells. Memórias do Instituto Oswaldo Cruz 112, 664673.10.1590/0074-02760170032CrossRefGoogle Scholar
Figueroa-Angulo, EE, Rendón-Gandarilla, FJ, Puente-Rivera, J, Calla-Choque, JS, Cárdenas-Guerra, RE, Ortega-López, J, Quintas-Granados, LI, Álvarez-Sánchez, ME and Arroyo, R (2012) The effects of environmental factors on the virulence of Trichomonas vaginalis. Microbes and Infection 14, 14111427.10.1016/j.micinf.2012.09.004CrossRefGoogle ScholarPubMed
Fiori, PL, Rappelli, P, Rocchigiani, AM and Cappuccinelli, P (1993) Trichomonas vaginalis haemolysis: evidence of functional pores formation on red cell membranes. FEMS Microbiology 109, 1318.10.1111/j.1574-6968.1993.tb06136.xCrossRefGoogle ScholarPubMed
Fiori, PL, Rappelli, P, Addis, MF, Sechi, A and Capuccinelli, P (1996) Trichomonas vaginalis haemolysis: pH regulates a contact-independent mechanism based on pore-forming proteins. Microbial Pathogens 20, 109118.10.1006/mpat.1996.0010CrossRefGoogle ScholarPubMed
Fiori, PL, Rappelli, P and Addis, MF (1999) The flagellated parasite Trichomonas vaginalis: new insights into cytopathogenicity mechanisms. Microbes and Infection 1, 149156.10.1016/S1286-4579(99)80006-9CrossRefGoogle ScholarPubMed
Gorrell, TE (1985) Effect of culture medium iron content on the biochemical composition and metabolism of Trichomonas vaginalis. Journal of Bacteriology 161, 12281230.10.1128/JB.161.3.1228-1230.1985CrossRefGoogle ScholarPubMed
Harlow, E and Lane, D (1988) Antibodies: A Laboratory Manual. New York: Cold Spring Harbor Laboratory, pp. 298300. ISBN 0-87969-314-2.Google Scholar
Hernández-Gutiérrez, R, Ortega-López, J and Arroyo, R (2003) A 39-kDa cysteine proteinase CP39 from Trichomonas vaginalis, which is negatively affected by iron may be involved in trichomonal cytotoxicity. Journal of Eukaryotic Microbiology 50(Suppl), 696698.10.1111/j.1550-7408.2003.tb00692.xCrossRefGoogle ScholarPubMed
Hernández-Gutiérrez, R, Avila González, L, Ortega-López, J, Cruz-Talonia, F, Gómez-Gutiérrez, G and Arroyo, R (2004) Trichomonas vaginalis: characterization of a 39-kDa cysteine proteinase found in patient vaginal secretions. Experimental Parasitology 107, 125135.10.1016/j.exppara.2004.05.004CrossRefGoogle ScholarPubMed
Hernández, HM, Marcet, R and Sarracent, J (2014) Biological roles of cysteine proteinases in the pathogenesis of Trichomonas vaginalis. Parasite 21, 54.10.1051/parasite/2014054CrossRefGoogle ScholarPubMed
Horváthová, L, Šafaríková, L, Basler, M, Hrdy, I, Campo, NB, Shin, JW, Huang, KY, Huang, PJ, Lin, R, Tang, P and Tachezy, J (2012) Transcriptomic identification of iron-regulated and iron-independent gene copies within the heavily duplicated Trichomonas vaginalis genome. Genome Biological Evolution 4, 10171029.10.1093/gbe/evs078CrossRefGoogle ScholarPubMed
Kummer, S, Hayes, GR, Gilbert, RO, Beach, DH, Lucas, JJ and Singh, BN (2008) Induction of human host cell apoptosis by Trichomonas vaginalis cysteine proteases is modulated by parasite exposure to iron. Microbial Pathogens 44, 197203.10.1016/j.micpath.2007.09.004CrossRefGoogle Scholar
Lehker, MW and Alderete, JF (1992) Iron regulates growth of Trichomonas vaginalis and the expression of immunogenic trichomonad proteins. Molecular Microbiology 6, 123132.10.1111/j.1365-2958.1992.tb00844.xCrossRefGoogle ScholarPubMed
Lehker, MW and Alderete, JF (2000) Biology of trichomonosis. Current Opinion in Infectious Diseases 13, 745.10.1097/00001432-200002000-00007CrossRefGoogle ScholarPubMed
León-Sicairos, CR, León-Félix, J and Arroyo, R (2004) Tvcp12: a novel Trichomonas vaginalis cathepsin L-like cysteine proteinase encoding-gene. Microbiology (Reading, England) 150, 11311138.10.1099/mic.0.26927-0CrossRefGoogle ScholarPubMed
Lubick, KJ and Burgess, DE (2004) Purification and analysis of a phospholipase A2-like lytic factor of Trichomonas vaginalis. Infection and Immunity 72, 12841290.10.1128/IAI.72.3.1284-1290.2004CrossRefGoogle ScholarPubMed
Madico, G, Quinn, TC, Rompalo, A, McKee, KT and Gaydós, CA (1998) Diagnosis of Trichomonas vaginalis infection by using vaginal swab samples. Journal of Clinical Microbiology 36, 32053210.10.1128/JCM.36.11.3205-3210.1998CrossRefGoogle ScholarPubMed
Mallinson, DJ, Lockwood, BC, Coombs, GH and North, MJ (1994) Identification and molecular cloning of four cysteine proteinase genes from the pathogenic protozoon Trichomonas vaginalis. Microbiology (Reading, England) 140, 27252735.10.1099/00221287-140-10-2725CrossRefGoogle ScholarPubMed
Mendoza-López, MR, Becerril-García, C, Fattel-Facenda, LV, Ávila-González, L, et al. (2000) CP30, A cysteine proteinase involved in Trichomonas vaginalis cytoadherence. Infection and Immunity 68, 49074912.CrossRefGoogle ScholarPubMed
Meza-Cervantez, P, González-Robles, A, Cárdenas-Guerra, RE, Ortega-López, J, Saavedra, E, Pineda, E and Arroyo, R (2011) Pyruvate:ferredoxin oxidoreductase (PFO) is a surface-associated cell-binding protein in Trichomonas vaginalis and is involved in trichomonal adherence to host cells. Microbiology (Reading, England) 157, 34693482.10.1099/mic.0.053033-0CrossRefGoogle ScholarPubMed
Miranda-Ozuna, JFT, Hernández-García, MS, Brieba, LG, Benítez-Cardoza, CG, Ortega-López, J, González-Robles, A and Arroyo, R (2016) The glycolytic enzyme triosephosphate isomerase of Trichomonas vaginalis is a surface-associated protein induced by glucose that functions as a laminin- and fibronectin-binding protein. Infection and Immunity 84, 28782894.10.1128/IAI.00538-16CrossRefGoogle ScholarPubMed
Miranda-Ozuna, JFT, Rivera-Rivas, LA, Cárdenas-Guerra, RE, Hernández-García, MS, Rodríguez-Cruz, S, González-Robles, A, Chávez-Munguía, B and Arroyo, R (2019) Glucose-restriction increases Trichomonas vaginalis cellular damage towards HeLa cells and proteolytic activity of cysteine proteinases (CPs), such as TvCP2. Parasitology 146, 11561166.10.1017/S0031182019000209CrossRefGoogle Scholar
Neale, KA and Alderete, JF (1990) Analysis of the proteinases of representative Trichomonas vaginalis isolates. Infection and Immunity 58, 157162.CrossRefGoogle ScholarPubMed
Ortíz-Estrada, G, Calderón-Salinas, V, Shibayama-Salas, M, León-Sicairos, N and De la Garza, M (2015) Binding and endocytosis of bovine hololactoferrin by the parasite Entamoeba histolytica. Biomedical Research International 2015, 375836.10.1155/2015/375836CrossRefGoogle ScholarPubMed
Pastorek, L, Sobol, M and Hozák, P (2016) Colocalization coefficients evaluating the distribution of molecular targets in microscopy methods based on pointed patterns. Histochemical Cell Biology 146, 391406.10.1007/s00418-016-1467-yCrossRefGoogle ScholarPubMed
Petrin, D, Delgaty, K, Bhatt, R and Garber, G (1998) Clinical and microbiological aspects of Trichomonas vaginalis. Clinical Microbiology Reviews 11, 300317.CrossRefGoogle ScholarPubMed
Puente-Rivera, J, Ramón-Luing, LA, Figueroa-Angulo, EE, Ortega-López, J and Arroyo, R (2014) Trichocystatin-2 (TC-2): an endogenous inhibitor of cysteine proteinases in Trichomonas vaginalis is associated with TvCP39. International Journal of Biochemistry and Cell Biology 54C, 255265.10.1016/j.biocel.2014.04.005CrossRefGoogle Scholar
Puente-Rivera, J, Villalpando, JL, Villalobos-Osnaya, A, Vázquez-Carrillo, LI, León-Avila, G, Ponce-Regalado, MD, López-Camarillo, C, Elizalde-Contreras, JM, Ruiz-May, E, Arroyo, R and Alvarez-Sánchez, ME (2017) The 50 kDa metalloproteinase TvMP50 is a zinc-mediated Trichomonas vaginalis virulence factor. Molecular Biochemical Parasitology 217, 3241.10.1016/j.molbiopara.2017.09.001CrossRefGoogle Scholar
Ramón-Luing, LA, Rendón-Gandarilla, FJ, Puente-Rivera, J, Ávila-González, L and Arroyo, R (2011) Identification and characterization of the immunogenic cytotoxic TvCP39 proteinase gene of Trichomonas vaginalis. International Journal of Biochemistry and Cell Biology 43, 15001511.CrossRefGoogle Scholar
Ramón-Luing, LA, Rendón-Gandarilla, FJ, Cárdenas-Guerra, RE, Rodríguez-Cabrera, N, Ortega-López, J, et al. (2010) Immunoproteomics of the active degradome to identify biomarkers for Trichomonas vaginalis. Proteomics 10, 435444.CrossRefGoogle ScholarPubMed
Rendón-Gandarilla, FJ, Ramón-Luing, LA, Ortega-López, J, Rosa de Andrade, I, Benchimol, M and Arroyo, R (2013) The TvLEGU-1, a legumain-like cysteine proteinase, plays a key role in Trichomonas vaginalis cytoadherence. Biomed Research International 2013, 561979. doi: 10.1155/2013/561979.CrossRefGoogle Scholar
Ryu, JS, Choi, HK, Min, DY, Ha, SE and Ahn, MH (2001) Effect of iron on the virulence of Trichomonas vaginalis. Journal of Parasitology 87, 457460.CrossRefGoogle ScholarPubMed
Sánchez-Rodríguez, DB, Ortega-López, J, Cárdenas-Guerra, RE, Reséndiz-Cardiel, G, Chávez-Munguía, B, Lagunes-Guillén, A and Arroyo, R (2018) Characterization of a novel endogenous cysteine proteinase inhibitor trichocystatin-3 (TC-3), localized on the surface of Trichomonas vaginalis. International Journal of Biochemistry and Cell Biology 102, 87100.CrossRefGoogle Scholar
Singh, M, Sharma, H and Singh, N (2007) Hydrogen peroxide induces apoptosis in HeLa cells through mitochondrial pathway. Mitochondrion 7, 367373.10.1016/j.mito.2007.07.003CrossRefGoogle ScholarPubMed
Solano-González, E, Burrola-Barraza, E, León-Sicairos, C, Ávila-González, L, Gutiérrez-Escolano, L, Ortega-López, J and Arroyo, R (2007) The trichomonad cysteine proteinase TvCP4 transcript contains an iron-responsive element. FEBS Letters 581, 29192928.CrossRefGoogle ScholarPubMed
Sommer, U, Costello, CE, Hayes, GR, Beach, DH, Gilbert, DO, et al. (2005) Identification of Trichomonas vaginalis cysteine proteases that induce apoptosis in human vaginal epithelial cells. Journal of Biological Chemistry 280, 28532860.10.1074/jbc.M501752200CrossRefGoogle ScholarPubMed
Torres-Romero, JC and Arroyo, R (2009) Responsiveness of Trichomonas vaginalis to iron concentrations: evidence for a post-transcriptional iron regulation by an IRE/IRP-like system. Infection Genetics and Evolution 9, 10651074.CrossRefGoogle ScholarPubMed
Tsai, CD, Liu, HS and Tai, JH (2002) Characterization of an iron-responsive promoter in the protozoan pathogen Trichomonas vaginalis. Journal of Biological Chemistry 277, 51535162.CrossRefGoogle ScholarPubMed
World Health Organization (2012) A baseline report on global sexually transmitted infection surveillance.Google Scholar
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