Hostname: page-component-8448b6f56d-c4f8m Total loading time: 0 Render date: 2024-04-19T05:01:41.679Z Has data issue: false hasContentIssue false
  • English
  • Français

Characteristic pro-inflammatory cytokines and host defence cathelicidin peptide produced by human monocyte-derived macrophages infected with Neospora caninum

Published online by Cambridge University Press:  24 November 2017

E. Boucher ,
M. Marin ,
R. Holani ,
M. Young-Speirs ,
D.M. Moore  and
E.R. Cobo Open the ORCID record for E.R. Cobo [Opens in a new window]
E. Boucher
Affiliation:
Bachelor of Health Sciences, Cumming School of Medicine, University of Calgary, Calgary, Canada
M. Marin
Affiliation:
National Research Council (CONICET), Balcarce, Argentina
R. Holani
Affiliation:
Production Animal Health, Faculty of Veterinary Medicine, University of Calgary, Calgary, Canada
M. Young-Speirs
Affiliation:
Bachelor of Health Sciences, Cumming School of Medicine, University of Calgary, Calgary, Canada
D.M. Moore
Affiliation:
National Research Council (CONICET), Balcarce, Argentina
E.R. Cobo*
Affiliation:
Production Animal Health, Faculty of Veterinary Medicine, University of Calgary, Calgary, Canada
*
Author for correspondence: Eduardo R. Cobo, E-mail: ecobo@ucalgary.ca
Get access Rights & Permissions [Opens in a new window]

Abstract

Neospora caninum is a coccidian intracellular protozoan capable of infecting a wide range of mammals, although severe disease is mostly reported in dogs and cattle. Innate defences triggered by monocytes/macrophages are key in the pathogenesis of neosporosis, as these cells are first-line defenders against intracellular infections. The aim of this study was to characterize infection and innate responses in macrophages infected with N. caninum using a well-known cell model to study macrophage functions (human monocyte THP-1 cells). Intracellular invasion of live tachyzoites occurred as fast as 4 h (confirmed with immunofluorescence microscopy using N. caninum-specific antibodies). Macrophages infected by N. caninum had increased expression of pro-inflammatory cytokines (TNFα, IL-1β, IL-8, IFNγ). Interestingly, N. caninum induced expression of host-defence peptides (cathelicidins), a mechanism of defence never reported for N. caninum infection in macrophages. The expression of cytokines and cathelicidins in macrophages invaded by N. caninum was mediated by mitogen-activated protein kinase (MEK 1/2). Secretion of such innate factors from N. caninum-infected macrophages reduced parasite internalization and promoted the secretion of pro-inflammatory cytokines in naïve macrophages. We concluded that rapid invasion of macrophages by N. caninum triggered protective innate defence mechanisms against intracellular pathogens.

Keywords

CathelicidinscytokinesinfectionmacrophagesmonocytesNeospora caninum
Type
Research Article
Information
Parasitology , Volume 145 , Issue 7 , June 2018 , pp. 871 - 884
Check for updates
Copyright
Copyright © Cambridge University Press 2017 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abe, C, Tanaka, S, Ihara, F and Nishikawa, Y (2014) Macrophage depletion prior to Neospora caninum infection results in severe neosporosis in mice. Clinical and Vaccine Immunology 21, 1185–1188.Google Scholar
Akira, S and Kishimoto, T (1997) NF-IL6 and NF-kappa B in cytokine gene regulation. Advances in Immunology 65, 1–46.CrossRefGoogle ScholarPubMed
Alessi, DR, Cuenda, A, Cohen, P, Dudley, DT and Saltiel, AR (1995) PD 098059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. The Journal of Biological Chemistry 270, 27489–27494.Google Scholar
Bustin, SA, Benes, V, Garson, JA, Hellemans, J, Huggett, J, Kubista, M, Mueller, R, Nolan, T, Pfaffl, MW, Shipley, GL, Vandesompele, J and Wittwer, CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clinical Chemistry 55, 611–622.Google Scholar
Butcher, BA, Kim, L, Johnson, PF and Denkers, EY (2001) Toxoplasma gondii tachyzoites inhibit proinflammatory cytokine induction in infected macrophages by preventing nuclear translocation of the transcription factor NF-kappa B. The Journal of Immunology 167, 2193–2201.Google Scholar
Butcher, BA, Kim, L, Panopoulos, AD, Watowich, SS, Murray, PJ and Denkers, EY (2005) Cutting edge: IL-10-independent STAT3 activation by Toxoplasma gondii mediates suppression of IL-12 and TNF-α in host macrophages. The Journal of Immunology 174, 3148–3152.CrossRefGoogle Scholar
Canton, GJ, Katzer, F, Benavides-Silvan, J, Maley, SW, Palarea-Albaladejo, J, Pang, Y, Smith, S, Bartley, PM, Rocchi, M, Innes, EA and Chianini, F (2013) Phenotypic characterisation of the cellular immune infiltrate in placentas of cattle following experimental inoculation with Neospora caninum in late gestation. Veterinary Research 44, 60.CrossRefGoogle ScholarPubMed
Carvalho, JV, Alves, CM, Cardoso, MR, Mota, CM, Barbosa, BF, Ferro, EA, Silva, NM, Mineo, TW, Mineo, JR and Silva, DA (2010) Differential susceptibility of human trophoblastic (BeWo) and uterine cervical (HeLa) cells to Neospora caninum infection. International Journal for Parasitology 40, 1629–1637.Google Scholar
Chanput, W, Mes, JJ and Wichers, HJ (2014) THP-1 cell line: an in vitro cell model for immune modulation approach. International Immunopharmacology 23, 37–45.Google Scholar
Clough, B, Wright, JD, Pereira, PM, Hirst, EM, Johnston, AC, Henriques, R and Frickel, EM (2016) K63-Linked ubiquitination targets Toxoplasma gondii for endo-lysosomal destruction in IFNgamma-stimulated human cells. PLoS Pathogens 12, e1006027.Google Scholar
Cobo, ER, Kissoon-Singh, V, Moreau, F and Chadee, K (2015) Colonic MUC2 mucin regulates the expression and antimicrobial activity of β-defensin 2. Mucosal Immunology 8, 1360–1372.Google Scholar
Collantes-Fernandez, E, Arrighi, RB, Alvarez-Garcia, G, Weidner, JM, Regidor-Cerrillo, J, Boothroyd, JC, Ortega-Mora, LM and Barragan, A (2012) Infected dendritic cells facilitate systemic dissemination and transplacental passage of the obligate intracellular parasite Neospora caninum in mice. PLoS ONE 7, e32123.CrossRefGoogle ScholarPubMed
Courret, N, Darche, S, Sonigo, P, Milon, G, Buzoni-Gatel, D and Tardieux, I (2006) CD11c- and CD11b-expressing mouse leukocytes transport single Toxoplasma gondii tachyzoites to the brain. Blood 107, 309–316.Google Scholar
Cuenda, A, Rouse, J, Doza, YN, Meier, R, Cohen, P, Gallagher, TF, Young, PR and Lee, JC (1995) SB 203580 is a specific inhibitor of a MAP kinase homologue which is stimulated by cellular stresses and interleukin-1. FEBS Letters 364, 229–233.Google Scholar
Daigneault, M, Preston, JA, Marriott, HM, Whyte, MK and Dockrell, DH (2010) The identification of markers of macrophage differentiation in PMA-stimulated THP-1 cells and monocyte-derived macrophages. PLoS ONE, 5, e8668.Google Scholar
da Silva, MV, Ferreira Franca, FB, Mota, CM, de Macedo, AG Jr, Ramos, EL, Santiago, FM, Mineo, JR and Mineo, TW (2017) Dectin-1 compromises innate responses and host resistance against Neospora caninum infection. Frontiers in Immunology 8, 245.Google Scholar
Dijkstra, T, Barkema, HW, Eysker, M, Hesselink, JW and Wouda, W (2002) Natural transmission routes of Neospora caninum between farm dogs and cattle. Veterinary Parasitology 105, 99–104.Google Scholar
Dion, S, Germon, S, Guiton, R, Ducournau, C and Dimier-Poisson, I (2011) Functional activation of T cells by dendritic cells and macrophages exposed to the intracellular parasite Neospora caninum. International Journal for Parasitology 41, 685–695.Google Scholar
Dubey, JP (2003) Review of Neospora caninum and neosporosis in animals. The Korean Journal of Parasitology 41, 1–16.Google Scholar
Dubey, JP, Hattel, AL, Lindsay, DS and Topper, MJ (1988) Neonatal Neospora caninum infection in dogs: isolation of the causative agent and experimental transmission. Journal of the American Veterinary Medical Association 193, 1259–1263.Google ScholarPubMed
Dubey, JP, Lindsay, DS, Anderson, ML, Davis, SW and Shen, SK (1992) Induced transplacental transmission of Neospora caninum in cattle. Journal of the American Veterinary Medical Association 201, 709–713.Google Scholar
Elsheikha, HM, McKinlay, CL, Elsaied, NA and Smith, PA (2013) Effects of Neospora caninum infection on brain microvascular endothelial cells bioenergetics. Parasites & Vectors 6, 24.Google Scholar
Flynn, RJ and Marshall, ES (2011) Parasite limiting macrophages promote IL-17 secretion in naive bovine CD4(+) T-cells during Neospora caninum infection. Veterinary Immunology Immunopathology 144, 423–429.Google Scholar
Futcher, B, Latter, GI, Monardo, P, McLaughlin, CS and Garrels, JI (1999) A sampling of the yeast proteome. Molecular and Cellular Biology 19, 7357–7368.Google Scholar
Gombart, AF, Borregaard, N and Koeffler, HP (2005) Human cathelicidin antimicrobial peptide (CAMP) gene is a direct target of the vitamin D receptor and is strongly up-regulated in myeloid cells by 1,25-dihydroxyvitamin D3. FASEB Journal 19, 1067–1077.CrossRefGoogle Scholar
Gygi, SP, Rochon, Y, Franza, BR and Aebersold, R (1999) Correlation between protein and mRNA abundance in yeast. Molecular and Cellular Biology 19, 1720–1730.Google Scholar
He, X, Gong, P, Wei, Z, Liu, W, Wang, W, Li, J, Yang, Z and Zhang, X (2017) Peroxisome proliferator-activated receptor-gamma-mediated polarization of macrophages in Neospora caninum infection. Experimental Parasitology 178, 37–44.CrossRefGoogle ScholarPubMed
Hecker, YP, Moore, DP, Quattrocchi, V, Regidor-Cerrillo, J, Verna, A, Leunda, MR, Morrell, E, Ortega-Mora, LM, Zamorano, P, Venturini, MC and Campero, CM (2013) Immune response and protection provided by live tachyzoites and native antigens from the NC-6 Argentina strain of Neospora caninum in pregnant heifers. Veterinary Parasitology 197, 436–446.CrossRefGoogle ScholarPubMed
Hemphill, A, Gottstein, B and Kaufmann, H (1996) Adhesion and invasion of bovine endothelial cells by Neospora caninum. Parasitology 112 (Pt 2), 183–197.Google Scholar
Hu, J, Roy, SK, Shapiro, PS, Rodig, SR, Reddy, SP, Platanias, LC, Schreiber, RD and Kalvakolanu, DV (2001) ERK1 and ERK2 activate CCAAAT/enhancer-binding protein-beta-dependent gene transcription in response to interferon-gamma. The Journal of Biological Chemistry 276, 287–297.Google Scholar
Hussen, J, Duvel, A, Sandra, O, Smith, D, Sheldon, IM, Zieger, P and Schuberth, HJ (2013) Phenotypic and functional heterogeneity of bovine blood monocytes. PLoS ONE 8, e71502.CrossRefGoogle ScholarPubMed
Li, H, Gade, P, Xiao, W and Kalvakolanu, DV (2007) The interferon signaling network and transcription factor C/EBP-beta. Cellular & Molecular Immunology 4, 407–418.Google Scholar
Lindsay, DS and Dubey, JP (1989) In vitro development of Neospora caninum (Protozoa: Apicomplexa) from dogs. The Journal of Parasitology 75, 163–165.Google Scholar
Lobato, J, Silva, D A, Mineo, TW, Amaral, JD, Segundo, GR, Costa-Cruz, JM, Ferreira, MS, Borges, AS and Mineo, JR (2006) Detection of immunoglobulin G antibodies to Neospora caninum in humans: high seropositivity rates in patients who are infected by human immunodeficiency virus or have neurological disorders. Clinical and Vaccine Immunology 13, 84–89.Google Scholar
Lv, Q, Li, J, Gong, P, Xing, S and Zhang, X (2010) Neospora caninum: in vitro culture of tachyzoites in MCF-7 human breast carcinoma cells. Experimental Parasitology 126, 536–539.Google Scholar
McCann, CM, Vyse, AJ, Salmon, RL, Thomas, D, Williams, DJ, McGarry, JW, Pebody, R and Trees, AJ (2008) Lack of serologic evidence of Neospora caninum in humans, England. Emerging Infectious Diseases 14, 978–980.Google Scholar
Mineo, TW, Oliveira, CJ, Silva, DA, Oliveira, LL, Abatepaulo, AR, Ribeiro, DP, Ferreira, BR, Mineo, JR and Silva, JS (2010) Neospora caninum excreted/secreted antigens trigger CC-chemokine receptor 5-dependent cell migration. International Journal for Parasitology 40, 797–805.CrossRefGoogle ScholarPubMed
Moore, DP, Echaide, I, Verna, AE, Leunda, MR, Cano, A, Pereyra, S, Zamorano, PI, Odeon, AC and Campero, CM (2011) Immune response to Neospora caninum native antigens formulated with immune stimulating complexes in calves. Veterinary Parasitology 175, 245–251.Google Scholar
Mota, CM, Oliveira, AC, Davoli-Ferreira, M, Silva, MV, Santiago, FM, Nadipuram, SM, Vashisht, AA, Wohlschlegel, JA, Bradley, PJ, Silva, JS, Mineo, JR and Mineo, TW (2016) Neospora caninum activates p38 MAPK as an evasion mechanism against innate immunity. Frontiers in Microbiology 7, 1456.Google Scholar
Natarajan, K, Singh, S, Burke, TR Jr, Grunberger, D and Aggarwal, BB (1996) Caffeic acid phenethyl ester is a potent and specific inhibitor of activation of nuclear transcription factor NF-kappa B. Proceedings of the National Academy of Sciences of the United States of America 93, 9090–9095.Google Scholar
Oshiro, LM, Motta-Castro, AR, Freitas, SZ, Cunha, RC, Dittrich, RL, Meirelles, AC and Andreotti, R (2015) Neospora caninum and Toxoplasma gondii serodiagnosis in human immunodeficiency virus carriers. Revista da Sociedade Brasileira de Medicina Tropical 48, 568–572.CrossRefGoogle ScholarPubMed
Park, EK, Jung, HS, Yang, HI, Yoo, MC, Kim, C and Kim, KS (2007) Optimized THP-1 differentiation is required for the detection of responses to weak stimuli. Inflammation Research 56, 45–50.CrossRefGoogle ScholarPubMed
Petersen, E, Lebech, M, Jensen, L, Lind, P, Rask, M, Bagger, P, Bjorkman, C and Uggla, A (1999) Neospora caninum infection and repeated abortions in humans. Emerging Infectious Diseases 5, 278–280.Google Scholar
Pollo-Oliveira, L, Post, H, Acencio, ML, Lemke, N, van den Toorn, H, Tragante, V, Heck, AJ, Altelaar, AF and Yatsuda, AP (2013) Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics. Parasites & Vectors 6, 335.Google Scholar
Rajapakse, R, Uring-Lambert, B, Andarawewa, KL, Rajapakse, RP, Abou-Bacar, A, Marcellin, L and Candolfi, E (2007) 1,25(OH)2D3 inhibits in vitro and in vivo intracellular growth of apicomplexan parasite Toxoplasma gondii. The Journal of Steroid Biochemistry and Molecular Biology 103, 811–814.Google Scholar
Ritter, DM, Kerlin, R, Sibert, G and Brake, D (2002) Immune factors influencing the course of infection with Neospora caninum in the murine host. The Journal of Parasitology 88, 271–280.Google Scholar
Rosbottom, A, Gibney, H, Kaiser, P, Hartley, C, Smith, RF, Robinson, R, Kipar, A and Williams, DJ (2011) Up regulation of the maternal immune response in the placenta of cattle naturally infected with Neospora caninum. PLoS ONE 6, e15799.CrossRefGoogle ScholarPubMed
Rosenberger, CM, Gallo, RL and Finlay, BB (2004) Interplay between antibacterial effectors: a macrophage antimicrobial peptide impairs intracellular Salmonella replication. Proceedings of the National Academy of Sciences of the United States of America 101, 2422–2427.Google Scholar
Roy, SK, Shuman, JD, Platanias, LC, Shapiro, PS, Reddy, SP, Johnson, PF and Kalvakolanu, DV (2005) A role for mixed lineage kinases in regulating transcription factor CCAAT/enhancer-binding protein-{beta}-dependent gene expression in response to interferon-{gamma}. The Journal of Biological Chemistry 280, 24462–24471.CrossRefGoogle ScholarPubMed
Shi, L, Kishore, R, McMullen, MR and Nagy, LE (2002) Lipopolysaccharide stimulation of ERK1/2 increases TNF-alpha production via Egr-1. American Journal of Physiology. Cell Physiology 282, C1205–C1211.CrossRefGoogle ScholarPubMed
Spano, A, Barni, S and Sciola, L (2013) PMA withdrawal in PMA-treated monocytic THP-1 cells and subsequent retinoic acid stimulation, modulate induction of apoptosis and appearance of dendritic cells. Cell Proliferation 46, 328–347.Google Scholar
Tanaka, T, Nagasawa, H, Fujisaki, K, Suzuki, N and Mikami, T (2000) Growth-inhibitory effects of interferon-gamma on Neospora caninum in murine macrophages by a nitric oxide mechanism. Parasitol Research 86, 768–771.Google Scholar
Tarique, AA, Logan, J, Thomas, E, Holt, PG, Sly, PD and Fantino, E (2015) Phenotypic, functional, and plasticity features of classical and alternatively activated human macrophages. American Journal of Respiratory Cell and Molecular Biology 53, 676–688.Google Scholar
Taubert, A, Krull, M, Zahner, H and Hermosilla, C (2006 a) Toxoplasma gondii and Neospora caninum infections of bovine endothelial cells induce endothelial adhesion molecule gene transcription and subsequent PMN adhesion. Veterinary Immunology and Immunopathology 112, 272–283.Google Scholar
Taubert, A, Zahner, H and Hermosilla, C (2006 b) Dynamics of transcription of immunomodulatory genes in endothelial cells infected with different coccidian parasites. Veterinary Parasitology 142, 214–222.Google Scholar
Tomasinsig, L, Pizzirani, C, Skerlavaj, B, Pellegatti, P, Gulinelli, S, Tossi, A, Di Virgilio, F and Zanetti, M (2008) The human cathelicidin LL-37 modulates the activities of the P2X7 receptor in a structure-dependent manner. The Journal of Biological Chemistry 283, 30471–30481.CrossRefGoogle Scholar
Tosh, KW, Mittereder, L, Bonne-Annee, S, Hieny, S, Nutman, TB, Singer, SM, Sher, A and Jankovic, D (2016) The IL-12 response of primary human dendritic cells and monocytes to Toxoplasma gondii is stimulated by phagocytosis of live parasites rather than host cell invasion. The Journal of Immunology 196, 345–356.Google Scholar
Tranas, J, Heinzen, RA, Weiss, LM and McAllister, MM (1999) Serological evidence of human infection with the protozoan Neospora caninum. Clinical and Diagnostic Laboratory Immunology 6, 765–767.Google Scholar
Tunev, SS, McAllister, MM, Anderson-Sprecher, RC and Weiss, LM (2002) Neospora caninum in vitro: evidence that the destiny of a parasitophorous vacuole depends on the phenotype of the progenitor zoite. The Journal of Parasitology 88, 1095–1099.Google Scholar
Uggla, A, Stenlund, S, Holmdahl, OJ, Jakubek, EB, Thebo, P, Kindahl, H and Bjorkman, C (1998) Oral Neospora caninum inoculation of neonatal calves. International Journal for Parasitology 28, 1467–1472.CrossRefGoogle ScholarPubMed
Valere, A, Garnotel, R, Villena, I, Guenounou, M, Pinon, JM and Aubert, D (2003) Activation of the cellular mitogen-activated protein kinase pathways ERK, P38 and JNK during Toxoplasma gondii invasion. Parasite 10, 59–64.Google Scholar
Wang, X, Gong, P, Zhang, X, Wang, J, Tai, L, Wang, X, Wei, Z, Yang, Y, Yang, Z, Li, J and Zhang, X (2017) NLRP3 inflammasome activation in murine macrophages caused by Neospora caninum infection. Parasites & Vectors 10, 266.CrossRefGoogle ScholarPubMed
Yamane, I, Kitani, H, Kokuho, T, Shibahara, T, Haritani, M, Hamaoka, T, Shimizu, S, Koiwai, M, Shimura, K and Yokomizo, Y (2000) The inhibitory effect of interferon gamma and tumor necrosis factor alpha on intracellular multiplication of Neospora caninum in primary bovine brain cells. The Journal Veterinary Medical Sciences 62, 347–351.CrossRefGoogle ScholarPubMed
Yuk, JM, Shin, DM, Lee, HM, Yang, CS, Jin, HS, Kim, KK, Lee, ZW, Lee, SH, Kim, JM and Jo, EK (2009) Vitamin D3 induces autophagy in human monocytes/macrophages via cathelicidin. Cell Host & Microbe 6, 231–243.Google Scholar
Supplementary material: Image

Boucher et al supplementary material

Figure S1

Download Boucher et al supplementary material(Image)
Image 33.7 MB
Supplementary material: File

Boucher et al supplementary material

Boucher et al supplementary material 1

Download Boucher et al supplementary material(File)
File 12.3 KB