Infections

Ehrlichia ruminantium uses its transmembrane protein Ape to adhere to host bovine aortic endothelial cells

10.24072/pcjournal.189 - Peer Community Journal, Volume 2 (2022), article no. e70.

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Ehrlichia ruminantium is an obligate intracellular bacterium, transmitted by ticks of the genus Amblyomma and responsible for heartwater, a disease of domestic and wild ruminants. High genetic diversity of E. ruminantium strains hampers the development of an effective vaccine against all strains present in the field. In order to develop strategies for the control of heartwater through both vaccine and alternative therapeutic approaches, it is important to first gain a better understanding of the early interaction of E. ruminantium and its host cell. Particularly, the mechanisms associated with bacterial adhesion remain to be elucidated. Herein, we studied the role of E. ruminantium membrane protein ERGA_CDS_01230 (UniProt Q5FFA9), a probable iron transporter, in the adhesion process to host bovine aortic endothelial cells (BAEC). The recombinant version of the protein ERGA_CDS_01230, successfully produced in the Leishmania tarentolae system, is O-glycosylated. Following in vitro culture of E. ruminantium in BAEC, the expression of CDS ERGA_CDS_01230 peaks at the extracellular infectious elementary body stages. This result suggest the likely involvement of ERGA_CDS_01230, named hereafter Ape for Adhesion protein of Ehrlichia, in the early interaction of E. ruminantium with its host cells. We showed using flow cytometry and scanning electron microscopy that beads coated with recombinant ERGA_CDS_01230 (rApe) adheres to BAEC. In addition, we also observed that rApe interacts with proteins of the cell lysate, membrane and organelle fractions. Additionally, enzymatic treatment degrading dermatan and chondroitin sulfates on the surface of BAEC is associated with a 50% reduction in the number of bacteria in the host cell after a developmental cycle, indicating that glycosaminoglycans seem to play a role in the adhesion of E. ruminantium to the host cell. Finally, Ape induces a humoral response in vaccinated animals. Globally, our work identifying the role of Ape in E. ruminantium adhesion to host cells makes it a gold vaccine candidate and represents a first step toward the understanding of the mechanisms of cell invasion by E. ruminantium.

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DOI: 10.24072/pcjournal.189
Pinarello, Valérie 1, 2; Bencurova, Elena 3; Marcelino, Isabel 4; Gros, Olivier 5; Puech, Carinne 2; Bhide, Mangesh 3, 6; Vachiery, Nathalie 2; Meyer, Damien F. 1, 2

1 CIRAD, UMR ASTRE, F-97170 – Petit-Bourg, Guadeloupe, France
2 ASTRE, CIRAD, INRAE, Univ Montpellier – Montpellier, France
3 Laboratory of biomedical microbiology and immunology, University of veterinary medicine and pharmacy in Kosice, Komenskeho 73 – Kosice, Slovakia
4 Institut Pasteur de la Guadeloupe – Les Abymes Cedex, Guadeloupe, France
5 Institut de Systématique, Évolution, Biodiversité (ISYEB), Muséum national d’Histoire naturelle, CNRS, Sorbonne Université, EPHE, Université des Antilles Campus de Fouillole – Pointe-à-Pitre, France
6 Institute of neuroimmunology, Slovak academy of sciences, Dubravska cesta 9 – Bratislava, Slovakia
License: CC-BY 4.0
Copyrights: The authors retain unrestricted copyrights and publishing rights
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     title = {<i>Ehrlichia ruminantium</i> uses its transmembrane protein {Ape} to adhere to host bovine aortic endothelial cells},
     journal = {Peer Community Journal},
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Pinarello, Valérie; Bencurova, Elena; Marcelino, Isabel; Gros, Olivier; Puech, Carinne; Bhide, Mangesh; Vachiery, Nathalie; Meyer, Damien F. Ehrlichia ruminantium uses its transmembrane protein Ape to adhere to host bovine aortic endothelial cells. Peer Community Journal, Volume 2 (2022), article  no. e70. doi : 10.24072/pcjournal.189. https://peercommunityjournal.org/articles/10.24072/pcjournal.189/

Peer reviewed and recommended by PCI : 10.24072/pci.infections.100003

Conflict of interest of the recommender and peer reviewers:
The recommender in charge of the evaluation of the article and the reviewers declared that they have no conflict of interest (as defined in the code of conduct of PCI) with the authors or with the content of the article.

[1] Allsopp, B. A. Natural history of Ehrlichia ruminantium, Veterinary Parasitology, Volume 167 (2010) no. 2-4, pp. 123-135 | DOI

[2] Aquino, R. S.; Lee, E. S.; Park, P. W. Diverse Functions of Glycosaminoglycans in Infectious Diseases, Progress in Molecular Biology and Translational Science, Elsevier, 2010, pp. 373-394 | DOI

[3] Aquino, R. S.; Teng, Y. H.-F.; Park, P. W. Glycobiology of syndecan-1 in bacterial infections, Biochemical Society Transactions, Volume 46 (2018) no. 2, pp. 371-377 | DOI

[4] Bartlett, A. H.; Park, P. W. Proteoglycans in host–pathogen interactions: molecular mechanisms and therapeutic implications, Expert Reviews in Molecular Medicine, Volume 12 (2010) | DOI

[5] Brown, R. N.; Romine, M. F.; Schepmoes, A. A.; Smith, R. D.; Lipton, M. S. Mapping the Subcellular Proteome of Shewanella oneidensis MR-1 using Sarkosyl-Based Fractionation and LC−MS/MS Protein Identification, Journal of Proteome Research, Volume 9 (2010) no. 9, pp. 4454-4463 | DOI

[6] Budachetri, K.; Teymournejad, O.; Lin, M.; Yan, Q.; Mestres-Villanueva, M.; Brock, G. N.; Rikihisa, Y. An Entry-Triggering Protein of Ehrlichia Is a New Vaccine Candidate against Tick-Borne Human Monocytic Ehrlichiosis, mBio, Volume 11 (2020) no. 4 | DOI

[7] Cangi, N.; Gordon, J. L.; Bournez, L.; Pinarello, V.; Aprelon, R.; Huber, K.; Lefrançois, T.; Neves, L.; Meyer, D. F.; Vachiéry, N. Recombination Is a Major Driving Force of Genetic Diversity in the Anaplasmataceae Ehrlichia ruminantium, Frontiers in Cellular and Infection Microbiology, Volume 6 (2016) | DOI

[8] Cangi, N.; Pinarello, V.; Bournez, L.; Lefrançois, T.; Albina, E.; Neves, L.; Vachiéry, N. Efficient high-throughput molecular method to detect Ehrlichia ruminantium in ticks, Parasites &amp; Vectors, Volume 10 (2017) no. 1 | DOI

[9] Cheng, Z.; Wang, X.; Rikihisa, Y. Regulation of Type IV Secretion Apparatus Genes during Ehrlichia chaffeensis Intracellular Development by a Previously Unidentified Protein, Journal of Bacteriology, Volume 190 (2008) no. 6, pp. 2096-2105 | DOI

[10] Delafuente, J.; Garciagarcia, J.; Barbet, A.; Blouin, E.; Kocan, K. Adhesion of outer membrane proteins containing tandem repeats of and species (Rickettsiales: Anaplasmataceae) to tick cells, Veterinary Microbiology, Volume 98 (2004) no. 3-4, pp. 313-322 | DOI

[11] de la Fuente, J.; Garcia-Garcia, J. C.; Blouin, E. F.; Kocan, K. M. Characterization of the functional domain of major surface protein 1a involved in adhesion of the rickettsia Anaplasma marginale to host cells, Veterinary Microbiology, Volume 91 (2003) no. 2-3, pp. 265-283 | DOI

[12] Deem, S. review of heartwater and the threat of introduction of Cowdria ruminantium and Amblyomma spp. ticks to the American mainland, J Zoo Wildl Med, Volume 29 (1998), pp. 109-113

[13] DeLano, W. The PyMOL Molecular Graphics, DeLano Scientific, San Carlos, CA, USA. (System.http://www.pymol.org)

[14] Dumler, J. S.; Barbet, A. F.; Bekker, C. P.; Dasch, G. A.; Palmer, G. H.; Ray, S. C.; Rikihisa, Y.; Rurangirwa, F. R. Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and 'HGE agent' as subjective synonyms of Ehrlichia phagocytophila., International Journal of Systematic and Evolutionary Microbiology, Volume 51 (2001) no. 6, pp. 2145-2165 | DOI

[15] Formosa-Dague, C.; Castelain, M.; Martin-Yken, H.; Dunker, K.; Dague, E.; Sletmoen, M. The Role of Glycans in Bacterial Adhesion to Mucosal Surfaces: How Can Single-Molecule Techniques Advance Our Understanding?, Microorganisms, Volume 6 (2018) no. 2 | DOI

[16] Frutos, R.; Viari, A.; Ferraz, C.; Bensaid, A.; Morgat, A.; Boyer, F.; Coissac, E.; Vachiéry, N.; Demaille, J.; Martinez, D. Comparative Genomics of Three Strains of Ehrlichia ruminantium, Annals of the New York Academy of Sciences, Volume 1081 (2006) no. 1, pp. 417-433 | DOI

[17] Frutos, R.; Viari, A.; Vachiery, N.; Boyer, F.; Martinez, D. Ehrlichia ruminantium: genomic and evolutionary features, Trends in Parasitology, Volume 23 (2007) no. 9, pp. 414-419 | DOI

[18] Gagoski, D.; Mureev, S.; Giles, N.; Johnston, W.; Dahmer-Heath, M.; Škalamera, D.; Gonda, T. J.; Alexandrov, K. Gateway-compatible vectors for high-throughput protein expression in pro- and eukaryotic cell-free systems, Journal of Biotechnology, Volume 195 (2015), pp. 1-7 | DOI

[19] García, B.; Merayo-Lloves, J.; Martin, C.; Alcalde, I.; Quirós, L. M.; Vazquez, F. Surface Proteoglycans as Mediators in Bacterial Pathogens Infections, Frontiers in Microbiology, Volume 7 (2016) | DOI

[20] Green, R. S.; Izac, J. R.; Naimi, W. A.; O'Bier, N.; Breitschwerdt, E. B.; Marconi, R. T.; Carlyon, J. A. Ehrlichia chaffeensis EplA Interaction With Host Cell Protein Disulfide Isomerase Promotes Infection, Frontiers in Cellular and Infection Microbiology, Volume 10 (2020) | DOI

[21] Hebert, K. S.; Seidman, D.; Oki, A. T.; Izac, J.; Emani, S.; Oliver, L. D.; Miller, D. P.; Tegels, B. K.; Kannagi, R.; Marconi, R. T.; Carlyon, J. A. Anaplasma marginale Outer Membrane Protein A Is an Adhesin That Recognizes Sialylated and Fucosylated Glycans and Functionally Depends on an Essential Binding Domain, Infection and Immunity, Volume 85 (2017) no. 3 | DOI

[22] Herron, M. J.; Nelson, C. M.; Larson, J.; Snapp, K. R.; Kansas, G. S.; Goodman, J. L. Intracellular Parasitism by the Human Granulocytic Ehrlichiosis Bacterium Through the P-Selectin Ligand, PSGL-1, Science, Volume 288 (2000) no. 5471, pp. 1653-1656 | DOI

[23] Hillman, R. D.; Baktash, Y. M.; Martinez, J. J. OmpA-mediated rickettsial adherence to and invasion of human endothelial cells is dependent upon interaction with α2β1 integrin, Cellular Microbiology, Volume 15 (2012) no. 5, pp. 727-741 | DOI

[24] Jonquieres, R.; Pizarro-Cerda, J.; Cossart, P. Synergy between the N- and C-terminal domains of InlB for efficient invasion of non-phagocytic cells by Listeria monocytogenes, Molecular Microbiology, Volume 42 (2001) no. 4, pp. 955-965 | DOI

[25] Kahlon, A.; Ojogun, N.; Ragland, S. A.; Seidman, D.; Troese, M. J.; Ottens, A. K.; Mastronunzio, J. E.; Truchan, H. K.; Walker, N. J.; Borjesson, D. L.; Fikrig, E.; Carlyon, J. A. Anaplasma phagocytophilum Asp14 Is an Invasin That Interacts with Mammalian Host Cells via Its C Terminus To Facilitate Infection, Infection and Immunity, Volume 81 (2013) no. 1, pp. 65-79 | DOI

[26] Kobayashi, K.; Kato, K.; Sugi, T.; Takemae, H.; Pandey, K.; Gong, H.; Tohya, Y.; Akashi, H. Plasmodium falciparum BAEBL Binds to Heparan Sulfate Proteoglycans on the Human Erythrocyte Surface, Journal of Biological Chemistry, Volume 285 (2010) no. 3, pp. 1716-1725 | DOI

[27] Leong, J. M.; Robbins, D.; Rosenfeld, L.; Lahiri, B.; Parveen, N. Structural Requirements for Glycosaminoglycan Recognition by the Lyme Disease Spirochete, Borrelia burgdorferi, Infection and Immunity, Volume 66 (1998) no. 12, pp. 6045-6048 | DOI

[28] Lin, M.; Liu, H.; Xiong, Q.; Niu, H.; Cheng, Z.; Yamamoto, A.; Rikihisa, Y. Ehrlichia secretes Etf-1 to induce autophagy and capture nutrients for its growth through RAB5 and class III phosphatidylinositol 3-kinase, Autophagy, Volume 12 (2016) no. 11, pp. 2145-2166 | DOI

[29] Lin, Y.-P.; Li, L.; Zhang, F.; Linhardt, R. J. Borrelia burgdorferi glycosaminoglycan-binding proteins: a potential target for new therapeutics against Lyme disease, Microbiology, Volume 163 (2017) no. 12, pp. 1759-1766 | DOI

[30] Lundberg, M.; Wikström, S.; Johansson, M. Cell surface adherence and endocytosis of protein transduction domains, Molecular Therapy, Volume 8 (2003) no. 1, pp. 143-150 | DOI

[31] Marcelino, I.; Colomé-Calls, N.; Holzmuller, P.; Lisacek, F.; Reynaud, Y.; Canals, F.; Vachiéry, N. Sweet and Sour Ehrlichia: Glycoproteomics and Phosphoproteomics Reveal New Players in Ehrlichia ruminantium Physiology and Pathogenesis, Frontiers in Microbiology, Volume 10 (2019) | DOI

[32] Marcelino, I.; Veríssimo, C.; Sousa, M. F.; Carrondo, M. J.; Alves, P. M. Characterization of Ehrlichia ruminantium replication and release kinetics in endothelial cell cultures, Veterinary Microbiology, Volume 110 (2005) no. 1-2, pp. 87-96 | DOI

[33] Martin, C.; Lozano-Iturbe, V.; Girón, R. M.; Vazquez-Espinosa, E.; Rodriguez, D.; Merayo-Lloves, J.; Vazquez, F.; Quirós, L. M.; García, B. Glycosaminoglycans are differentially involved in bacterial binding to healthy and cystic fibrosis lung cells, Journal of Cystic Fibrosis, Volume 18 (2019) no. 3 | DOI

[34] Martinez, D.; Coisne, S.; Sheikboudou, C.; Jongejan, F. Détection d’anticorps contre Cowdria ruminantium; dans le sérum de ruminants domestiques par ELISA indirect, Revue d’élevage et de médecine vétérinaire des pays tropicaux, Volume 46 (1993) no. 1-2, pp. 115-120 | DOI

[35] Martinez, E.; Cantet, F.; Fava, L.; Norville, I.; Bonazzi, M. Identification of OmpA, a Coxiella burnetii Protein Involved in Host Cell Invasion, by Multi-Phenotypic High-Content Screening, PLoS Pathogens, Volume 10 (2014) no. 3 | DOI

[36] Martinez, J. J.; Seveau, S.; Veiga, E.; Matsuyama, S.; Cossart, P. Ku70, a Component of DNA-Dependent Protein Kinase, Is a Mammalian Receptor for Rickettsia conorii, Cell, Volume 123 (2005) no. 6, pp. 1013-1023 | DOI

[37] Miller, E. M.; Nickoloff, J. A. Escherichia coli Electrotransformation, Electroporation Protocols for Microorganisms, Humana Press, New Jersey, pp. 105-114 | DOI

[38] Mohan Kumar, D.; Yamaguchi, M.; Miura, K.; Lin, M.; Los, M.; Coy, J. F.; Rikihisa, Y. Ehrlichia chaffeensis Uses Its Surface Protein EtpE to Bind GPI-Anchored Protein DNase X and Trigger Entry into Mammalian Cells, PLoS Pathogens, Volume 9 (2013) no. 10 | DOI

[39] Moumène, A.; Gonzalez-Rizzo, S.; Lefrançois, T.; Vachiéry, N.; Meyer, D. F. Iron Starvation Conditions Upregulate Ehrlichia ruminantium Type IV Secretion System, tr1 Transcription Factor and map1 Genes Family through the Master Regulatory Protein ErxR, Frontiers in Cellular and Infection Microbiology, Volume 7 (2018) | DOI

[40] Moumène, A.; Marcelino, I.; Ventosa, M.; Gros, O.; Lefrançois, T.; Vachiéry, N.; Meyer, D. F.; Coelho, A. V. Proteomic Profiling of the Outer Membrane Fraction of the Obligate Intracellular Bacterial Pathogen Ehrlichia ruminantium, PLOS ONE, Volume 10 (2015) no. 2 | DOI

[41] Moumène, A.; Meyer, D. F. Ehrlichia's molecular tricks to manipulate their host cells, Microbes and Infection, Volume 18 (2016) no. 3, pp. 172-179 | DOI

[42] Niemann, H. H.; Schubert, W.-D.; Heinz, D. W. Adhesins and invasins of pathogenic bacteria: a structural view, Microbes and Infection, Volume 6 (2004) no. 1, pp. 101-112 | DOI

[43] Noroy, C.; Lefrançois, T.; Meyer, D. F. Searching algorithm for Type IV effector proteins (S4TE) 2.0: Improved tools for Type IV effector prediction, analysis and comparison in proteobacteria, PLOS Computational Biology, Volume 15 (2019) no. 3 | DOI

[44] Ojogun, N.; Kahlon, A.; Ragland, S. A.; Troese, M. J.; Mastronunzio, J. E.; Walker, N. J.; VieBrock, L.; Thomas, R. J.; Borjesson, D. L.; Fikrig, E.; Carlyon, J. A. Anaplasma phagocytophilum Outer Membrane Protein A Interacts with Sialylated Glycoproteins To Promote Infection of Mammalian Host Cells, Infection and Immunity, Volume 80 (2012) no. 11, pp. 3748-3760 | DOI

[45] Perez, J.-M.; Martinez, D.; Sheikboudou, C.; Jongejan, F.; Bensaid, A. Characterization of variable immunodominant antigens of Cowdria ruminantium by ELISA and immunoblots, Parasite Immunology, Volume 20 (1998) no. 12, pp. 613-622 | DOI

[46] Pizarro-Cerdá, J.; Cossart, P. Bacterial Adhesion and Entry into Host Cells, Cell, Volume 124 (2006) no. 4, pp. 715-727 | DOI

[47] Pruneau, L.; Emboulé, L.; Gely, P.; Marcelino, I.; Mari, B.; Pinarello, V.; Sheikboudou, C.; Martinez, D.; Daigle, F.; Lefrançois, T.; Meyer, D. F.; Vachiery, N. Global gene expression profiling of Ehrlichia ruminantium at different stages of development, FEMS Immunology &amp; Medical Microbiology, Volume 64 (2012) no. 1, pp. 66-73 | DOI

[48] Rajas, O.; Quirós, L. M.; Ortega, M.; Vazquez-Espinosa, E.; Merayo-Lloves, J.; Vazquez, F.; García, B. Glycosaminoglycans are involved in bacterial adherence to lung cells, BMC Infectious Diseases, Volume 17 (2017) no. 1 | DOI

[49] Reneer, D. V.; Troese, M. J.; Huang, B.; Kearns, S. A.; Carlyon, J. A. Anaplasma phagocytophilum PSGL-1-independent infection does not require Syk and leads to less efficient AnkA delivery, Cellular Microbiology, Volume 10 (2008) no. 9, pp. 1827-1838 | DOI

[50] Rikihisa, Y. Role and Function of the Type IV Secretion System in Anaplasma and Ehrlichia Species, Current Topics in Microbiology and Immunology, Springer International Publishing, Cham, 2017, pp. 297-321 | DOI

[51] Sava, I. G.; Zhang, F.; Toma, I.; Theilacker, C.; Li, B.; Baumert, T. F.; Holst, O.; Linhardt, R. J.; Huebner, J. Novel Interactions of Glycosaminoglycans and Bacterial Glycolipids Mediate Binding of Enterococci to Human Cells, Journal of Biological Chemistry, Volume 284 (2009) no. 27, pp. 18194-18201 | DOI

[52] Teymournejad, O.; Lin, M.; Rikihisa, Y. Ehrlichia chaffeensis and Its Invasin EtpE Block Reactive Oxygen Species Generation by Macrophages in a DNase X-Dependent Manner, mBio, Volume 8 (2017) no. 6 | DOI

[53] Tiwari, V.; Maus, E.; Sigar, I. M.; Ramsey, K. H.; Shukla, D. Role of heparan sulfate in sexually transmitted infections, Glycobiology, Volume 22 (2012) no. 11, pp. 1402-1412 | DOI

[54] Truttmann, M. C.; Rhomberg, T. A.; Dehio, C. Combined action of the type IV secretion effector proteins BepC and BepF promotes invasome formation of Bartonella henselae on endothelial and epithelial cells, Cellular Microbiology, Volume 13 (2010) no. 2, pp. 284-299 | DOI

[55] Waterhouse, A.; Bertoni, M.; Bienert, S.; Studer, G.; Tauriello, G.; Gumienny, R.; Heer, F. T.; de Beer, T. A. P.; Rempfer, C.; Bordoli, L.; Lepore, R.; Schwede, T. SWISS-MODEL: homology modelling of protein structures and complexes, Nucleic Acids Research, Volume 46 (2018) no. W1 | DOI

[56] Yan, Q.; Lin, M.; Huang, W.; Teymournejad, O.; Johnson, J. M.; Hays, F. A.; Liang, Z.; Li, G.; Rikihisa, Y. Ehrlichia type IV secretion system effector Etf-2 binds to active RAB5 and delays endosome maturation, Proceedings of the National Academy of Sciences, Volume 115 (2018) no. 38 | DOI

[57] Yang, J.; Zhang, Y. I-TASSER server: new development for protein structure and function predictions, Nucleic Acids Research, Volume 43 (2015) no. W1 | DOI

[58] Yu, C.-S.; Chen, Y.-C.; Lu, C.-H.; Hwang, J.-K. Prediction of protein subcellular localization, Proteins: Structure, Function, and Bioinformatics, Volume 64 (2006) no. 3, pp. 643-651 | DOI

[59] Zhou, X.; Zheng, W.; Li, Y.; Pearce, R.; Zhang, C.; Bell, E. W.; Zhang, G.; Zhang, Y. I-TASSER-MTD: a deep-learning-based platform for multi-domain protein structure and function prediction, Nature Protocols, Volume 17 (2022) no. 10, pp. 2326-2353 | DOI

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