Abstract
Naegleria fowleri is a protozoan that invades the central nervous system and causes primary amoebic meningoencephalitis. It has been reported that N. fowleri induces an important inflammatory response during the infection. In the present study, we evaluated the roles of Toll-like receptors in the recognition of N. fowleri trophozoites by human mucoepithelial cells, analyzing the expression and production of innate immune response mediators. After amoebic interactions with NCI-H292 cells, the expression and production levels of IL-8, TNF-α, IL-1β, and human beta defensin-2 were evaluated by RT-PCR, ELISA, immunofluorescence, and dot blot assays, respectively. To determine whether the canonical signaling pathways were engaged, we used different inhibitors, namely, IMG-2005 for MyD88 and BAY 11-7085 for the nuclear factor NFkB. Our results showed that the expression and production of the pro-inflammatory cytokines and beta defensin-2 were induced by N. fowleri mainly through the canonical TLR4 pathway in a time-dependent manner.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abhyankar MM, Noor Z, Tomai MA, Elvecrog J, Fox CB, Petri WA Jr (2017) Nanoformulation of synergistic TLR ligands to enhance vaccination against Entamoeba histolytica. Vaccine 35(6):916–922. https://doi.org/10.1016/j.vaccine.2016.12.057
Akira S, Takeda K (2004) Toll-like receptor signalling. Nat Rev Immunol 4(7):499–511. https://doi.org/10.1038/nri1391
Arbibe L et al (2000) Toll-like receptor 2-mediated NF-kappa B activation requires a Rac1-dependent pathway. Nat Immunol 1(6):533–540. https://doi.org/10.1038/82797
Barton GM, Medzhitov R (2003) Toll-like receptor signaling pathways. Science 300(5625):1524–1525. https://doi.org/10.1126/science.1085536
Becker MN, Diamond G, Verghese MW, Randell SH (2000) CD14-dependent lipopolysaccharide-induced beta-defensin-2 expression in human tracheobronchial epithelium. J Biol Chem 275(38):29731–29736. https://doi.org/10.1074/jbc.M000184200
Cario E, Rosenberg IM, Brandwein SL, Beck PL, Reinecker HC, Podolsky DK (2000) Lipopolysaccharide activates distinct signaling pathways in intestinal epithelial cell lines expressing Toll-like receptors. J Immunol 164(2):966–972. https://doi.org/10.4049/jimmunol.164.2.966
Cervantes-Sandoval I, Serrano-Luna J, Garcia-Latorre E, Tsutsumi V, Shibayama M (2008a) Mucins in the host defence against Naegleria fowleri and mucinolytic activity as a possible means of evasion. Microbiology 154(Pt 12):3895–3904. https://doi.org/10.1099/mic.0.2008/019380-0
Cervantes-Sandoval I, Serrano-Luna J, García-Latorre E, Tsutsumi V, Shibayama M (2008b) Characterization of brain inflammation during primary amoebic meningoencephalitis. Parasitol Int 57(3):307–313. https://doi.org/10.1016/j.parint.2008.01.006
Cervantes-Sandoval I, Serrano-Luna Jde J, Meza-Cervantez P, Arroyo R, Tsutsumi V, Shibayama M (2009) Naegleria fowleri induces MUC5AC and pro-inflammatory cytokines in human epithelial cells via ROS production and EGFR activation. Microbiology 155(Pt 11):3739–3747. https://doi.org/10.1099/mic.0.030635-0
Cervantes-Sandoval I, Serrano-Luna J, Pacheco-Yepez J, Silva-Olivares A, Tsutsumi V, Shibayama M (2010) Differences between Naegleria fowleri and Naegleria gruberi in expression of mannose and fucose glycoconjugates. Parasitol Res 106(3):695–701. https://doi.org/10.1007/s00436-010-1727-z
Coban C et al (2007) Pathological role of Toll-like receptor signaling in cerebral malaria. Int Immunol 19(1):67–79. https://doi.org/10.1093/intimm/dxl123
Coughlin SR (2000) Thrombin signalling and protease-activated receptors. Nature 407(6801):258–264. https://doi.org/10.1038/35025229
Cox FE (2002) History of human parasitology. Clin Microbiol Rev 15(4):595–612
Cuenda A, Rousseau S (2007) p38 MAP-kinases pathway regulation, function and role in human diseases. Biochim Biophys Acta 1773(8):1358–1375. https://doi.org/10.1016/j.bbamcr.2007.03.010
de Veer MJ et al (2003) MyD88 is essential for clearance of Leishmania major: possible role for lipophosphoglycan and Toll-like receptor 2 signaling. Eur J Immunol 33(10):2822–2831. https://doi.org/10.1002/eji.200324128
de Zoete MR, Bouwman LI, Keestra AM, van Putten JP (2011) Cleavage and activation of a Toll-like receptor by microbial proteases. Proc Natl Acad Sci U S A 108(12):4968–4973. https://doi.org/10.1073/pnas.1018135108
Derda M et al (2016) Acanthamoeba infection in lungs of mice expressed by toll-like receptors (TLR2 and TLR4). Exp Parasitol 165:30–34. https://doi.org/10.1016/j.exppara.2016.02.012
Drennan MB et al (2005) The induction of a type 1 immune response following a Trypanosoma brucei infection is MyD88 dependent. J Immunol 175(4):2501–2509. https://doi.org/10.4049/jimmunol.175.4.2501
Erridge C, Kennedy S, Spickett CM, Webb DJ (2008) Oxidized phospholipid inhibition of toll-like receptor (TLR) signaling is restricted to TLR2 and TLR4: roles for CD14, LPS-binding protein, and MD2 as targets for specificity of inhibition. J Biol Chem 283(36):24748–24759. https://doi.org/10.1074/jbc.M800352200
Faure E, Thomas L, Xu H, Medvedev A, Equils O, Arditi M (2001) Bacterial lipopolysaccharide and IFN-gamma induce Toll-like receptor 2 and Toll-like receptor 4 expression in human endothelial cells: role of NF-kappa B activation. J Immunol 166(3):2018–2024
Flo TH et al (2000) Human toll-like receptor 2 mediates monocyte activation by Listeria monocytogenes, but not by group B streptococci or lipopolysaccharide. J Immunol 164(4):2064–2069. https://doi.org/10.4049/jimmunol.164.4.2064
Froy O (2005) Regulation of mammalian defensin expression by Toll-like receptor-dependent and independent signalling pathways. Cell Microbiol 7(10):1387–1397. https://doi.org/10.1111/j.1462-5822.2005.00590.x
Galván-Moroyoqui JM, Del Carmen Domínguez-Robles M, Meza I (2011) Pathogenic bacteria prime the induction of toll-like receptor signalling in human colonic cells by the Gal/GalNAc lectin carbohydrate recognition domain of Entamoeba histolytica. Int J Parasitol 41(10):1101–1112. https://doi.org/10.1016/j.ijpara.2011.06.003
Han KL, Lee HJ, Shin MH, Shin HJ, Im KI, Park SJ (2004) The involvement of an integrin-like protein and protein kinase C in amoebic adhesion to fibronectin and amoebic cytotoxicity. Parasitol Res 94(1):53–60. https://doi.org/10.1007/s00436-004-1158-9
Jono H et al (2002) Transforming growth factor-beta-Smad signaling pathway cooperates with NF-kappa B to mediate nontypeable Haemophilus influenzae-induced MUC2 mucin transcription. J Biol Chem 277(47):45547–45557. https://doi.org/10.1074/jbc.M206883200
Kim JH et al (2012a) Functional roles of mannose-binding protein in the adhesion, cytotoxicity and phagocytosis of Acanthamoeba castellanii. Exp Parasitol 132(2):287–292. https://doi.org/10.1016/j.exppara.2012.08.007
Kim JH, Sohn HJ, Lee SH, Kwon D, Shin HJ (2012b) Induction of interleukin-8 by Naegleria fowleri lysates requires activation of extracellular signal-regulated kinase in human astroglial cells. Parasitol Res. https://doi.org/10.1007/s00436-012-2872-3
Kim JH et al (2013) Vaccination with lentiviral vector expressing the nfa1 gene confers a protective immune response to mice infected with Naegleria fowleri. Clin Vaccine Immunol 20(7):1055–1060. https://doi.org/10.1128/CVI.00210-13
Kim JH et al (2016) NLRP-3 inflammasome activation in THP-1 target cells triggered by pathogenic Naegleria fowleri. Infect Immunol. https://doi.org/10.1128/IAI.00275-16
Lee YJ et al (2011) Naegleria fowleri lysate induces strong cytopathic effects and pro-inflammatory cytokine release in rat microglial cells. Korean J Parasitol 49(3):285–290. https://doi.org/10.3347/kjp.2011.49.3.285
Lehnardt S (2010) Innate immunity and neuroinflammation in the CNS: the role of microglia in Toll-like receptor-mediated neuronal injury. Glia 58(3):253–263. https://doi.org/10.1002/glia.20928
Loharungsikul S et al (2008) Expression of toll-like receptors on antigen-presenting cells in patients with falciparum malaria. Acta Trop 105(1):10–15. https://doi.org/10.1016/j.actatropica.2007.08.002
Maldonado-Bernal C et al (2005) The innate immune response to Entamoeba histolytica lipopeptidophosphoglycan is mediated by toll-like receptors 2 and 4. Parasite Immunol 27(4):127–137. https://doi.org/10.1111/j.1365-3024.2005.00754.x
McGuinness DH, Dehal PK, Pleass RJ (2003) Pattern recognition molecules and innate immunity to parasites. Trends Parasitol 19(7):312–319. https://doi.org/10.1016/S1471-4922(03)00123-5
Mishra BB, Gundra UM, Teale JM (2009) Toll-like receptors in CNS parasitic infections. Curr Top Microbiol Immunol 336:83–104. https://doi.org/10.1007/978-3-642-00549-7_5
Mockenhaupt FP et al (2006) Toll-like receptor (TLR) polymorphisms in African children: common TLR-4 variants predispose to severe malaria. J Commun Dis 38(3):230–245
Moncada DM, Kammanadiminti SJ, Chadee K (2003) Mucin and toll-like receptors in host defense against intestinal parasites. Trends Parasitol 19(7):305–311. https://doi.org/10.1016/S1471-4922(03)00122-3
Muzio M et al (2000) Differential expression and regulation of toll-like receptors (TLR) in human leukocytes: selective expression of TLR3 in dendritic cells. J Immunol 164(11):5998–6004. https://doi.org/10.4049/jimmunol.164.11.5998
O’Neill LA, Bowie AG (2007) The family of five: TIR-domain-containing adaptors in toll-like receptor signalling. Nat Rev Immunol 7(5):353–364. https://doi.org/10.1038/nri2079
Oh YH et al (2005) Cytopathic changes and pro-inflammatory cytokines induced by Naegleria fowleri trophozoites in rat microglial cells and protective effects of an anti-Nfa1 antibody. Parasite Immunol 27(12):453–459. https://doi.org/10.1111/j.1365-3024.2005.00799.x
Palsson-McDermott EM, O’Neill LA (2007) Building an immune system from nine domains. Biochem Soc Trans 35(Pt 6):1437–1444. https://doi.org/10.1042/BST0351437
Poltorak A et al (1998) Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 282(5396):2085–2088
Ren MY, Wu XY (2011) Toll-like receptor 4 signalling pathway activation in a rat model of Acanthamoeba Keratitis. Parasite Immunol 33(1):25–33. https://doi.org/10.1111/j.1365-3024.2010.01247.x
Ren M, Gao L, Wu X (2010) TLR4: the receptor bridging Acanthamoeba challenge and intracellular inflammatory responses in human corneal cell lines. Immunol Cell Biol 88(5):529–536. https://doi.org/10.1038/icb.2010.6
Rivas-Santiago B, Sada E, Tsutsumi V, Aguilar-Leon D, Contreras JL, Hernandez-Pando R (2006) beta-Defensin gene expression during the course of experimental tuberculosis infection. J Infect Dis 194(5):697–701. https://doi.org/10.1086/506454
Rojas-Hernandez S, Jarillo-Luna A, Rodriguez-Monroy M, Moreno-Fierros L, Campos-Rodriguez R (2004) Immunohistochemical characterization of the initial stages of Naegleria fowleri meningoencephalitis in mice. Parasitol Res 94(1):31–36. https://doi.org/10.1007/s00436-004-1177-6
Schmeck B et al (2006) Pneumococci induced TLR- and Rac1-dependent NF-kappaB-recruitment to the IL-8 promoter in lung epithelial cells. Am J Phys Lung Cell Mol Phys 290(4):L730–L737. https://doi.org/10.1152/ajplung.00271.2005
Sedger LM, McDermott MF (2014) TNF and TNF-receptors: from mediators of cell death and inflammation to therapeutic giants - past, present and future. Cytokine Growth Factor Rev 25(4):453–472. https://doi.org/10.1016/j.cytogfr.2014.07.016
Serrano-Luna J, Pina-Vázquez C, Reyes-López M, Ortíz-Estrada G, de la Garza M (2013) Proteases from Entamoeba spp. and pathogenic free-living amoebae as virulence factors. J Trop Med 2013:890603. https://doi.org/10.1155/2013/890603
Shaykhiev R, Sierigk J, Herr C, Krasteva G, Kummer W, Bals R (2010) The antimicrobial peptide cathelicidin enhances activation of lung epithelial cells by LPS. FASEB J 24(12):4756–4766. https://doi.org/10.1096/fj.09-151332
Stamm LV, Drapp RL (2014) TLR2 and TLR4 mediate the TNFalpha response to Vibrio vulnificus biotype 1. Pathog Dis 71(3):357–361. https://doi.org/10.1111/2049-632X.12154
Sugita Y et al (1999) Primary amebic meningoencephalitis due to Naegleria fowleri: an autopsy case in Japan. Pathol Int 49(5):468–470. https://doi.org/10.1046/j.1440-1827.1999.00893.x
Valenzuela G, Lopez-Corella E, De Jonckheere JF (1984) Primary amoebic meningoencephalitis in a young male from northwestern Mexico. Trans R Soc Trop Med Hyg 78(4):558–559
Visintin A, Mazzoni A, Spitzer JH, Wyllie DH, Dower SK, Segal DM (2001) Regulation of toll-like receptors in human monocytes and dendritic cells. J Immunol 166(1):249–255
Visvesvara GS (2013) Infections with free-living amebae. Handb Clin Neurol 114:153–168. https://doi.org/10.1016/B978-0-444-53490-3.00010-8
Wojtkowiak-Giera A et al (2016) Toll-like receptors in the brain of mice following infection with Acanthamoeba spp. Parasitol Res 115(11):4335–4344. https://doi.org/10.1007/s00436-016-5217-9
Yoder JS et al (2012) Primary amebic meningoencephalitis deaths associated with sinus irrigation using contaminated tap water. Clin Infect Dis 55(9):e79–e85. https://doi.org/10.1093/cid/cis626
Acknowledgements
We are grateful to Angélica Silva-Olivares for her technical assistance in the amoeba culture and Jaime Escobar-Herrera for his technical assistance with confocal microscopy.
Funding
This work was supported by CONACyT grant No. 237523.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Rights and permissions
About this article
Cite this article
Martínez-Castillo, M., Santos-Argumedo, L., Galván-Moroyoqui, J.M. et al. Toll-like receptors participate in Naegleria fowleri recognition. Parasitol Res 117, 75–87 (2018). https://doi.org/10.1007/s00436-017-5666-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-017-5666-9