Biological effects of the EVs released by different
Abstract
Leishmania spp. release extracellular vesicles (EVs) containing parasite molecules, including several antigens and virulence factors. These EVs can interact with the host cells, such as immune cells, contributing to the parasite–host relationship. Studies have demonstrated that Leishmania-EVs can promote infection in experimental models and modulate the immune response. Although the immunomodulatory effect has been demonstrated, Leishmania-EVs can deliver parasite antigens and therefore have the potential for use as a new diagnostic tool and development of new therapeutic and vaccine approaches. This review aims to bring significant advances in the field of extracellular vesicles and Leishmania, focusing on their role in the cells of the immune system.
Keywords
- extracellular vesicles
- exosomes
- microvesicles
- Leishmania
- immune response
- leishmaniasis
1. Introduction
The host–parasite communication and the parasite’s intercellular interactions are crucial in the life cycle of the
Several works have demonstrated that
2. Extracellular vesicles (EVs): an overview
EVs can be detected in body fluids, including urine, saliva, blood, plasma, amniotic fluid, breast milk, ascites, synovial fluid, and cerebrospinal fluid [7, 12]. Structurally, they present a spherical shape with a double layer composed of lipids and proteins and can be filled with biomolecules from the cell of origin [13]. EVs are classified based on their biogenesis, composition, and size, namely—exosomes, microvesicles (MVs), and apoptotic bodies (ABs) [8, 13]. Although MVs and exosomes show structural similarities, they are different in size, content, lipid composition, and biogenesis [7]. ABs are released by apoptotic cells and have specific characteristics [12] that will not be covered in this review.
Exosomes present sizes between 20 and 100 nm [14]. They are formed by the internal invagination of the endosomal membrane, originating the multivesicular bodies (MVBs) [8]. After maturation, exosomes are secreted by exocytosis via fusion of MBVs with the cell surface, or they may be digested by lysosomes [14, 15]. Exosomes are rich in lipids (mainly phosphatidylserine, cholesterol, and ceramides), nucleic acids, and proteins [8]. In addition, proteins such as endosomal sorting complexes required for transport (ESCRT), Alix, tumor susceptibility gene 101 (TSG101), heat shock cognate 70 (HSC70), HSP90β, HSP60 and HSP70, proteins from the annexin family, and tetraspanins (cluster of differentiation 63 - CD63, CD9, CD81, and CD82) participate in the process of formation of exosomes [8, 16]. These molecules are increased in exosomes, but they are not exclusive markers of these EVs types [7].
MVs are a group of EVs with a diameter between 100 and 1,000 nm [7]. They are originated from the protrusion of the cytoplasmic membrane, and they can carry molecules of cell surface such as membrane receptors, integrins, adhesins, and others [8]. Some studies have shown that structures such as actin and microtubules (cytoskeleton), kinesins and myosins, and soluble NSF attachment receptors (SNAREs) play a role in the formation of MVs [17]. However, the molecular pathway is not well understood [8, 13, 18], and specific markers of MVs have not yet been described. The releasing of MVs and exosomes occurs under physiological cell conditions, but the quantity and content can be altered after stimuli, such as low oxygen and nitrogen content, oxidative stress, among others [4, 5, 19].
Different vesicle isolation techniques have been performed; however, centrifugation/ultracentrifugation and size exclusion chromatography are the most commonly used [7]. Flow cytometry, Western blotting, nanoparticle tracking technique (NTA), mass spectrometry, and electron microscopy have been used to quantify and better characterize the isolated EVs (exosomes and/or MVs) [7]. The inclusion of new methodologies and the discovering of specific EVs markers will bring a new perspective to understand the role of these nanoparticles in the biology and the pathophysiology of several diseases. In addition, there is a great expectation of the applications of EVs in diagnostics, treatments, and vaccine development.
Currently, there is a consensus that EVs play an important role in cell–cell communication being a vehicle for transporting molecules between cells, even cross-kingdom [8, 18, 20]. The effects on the recipient cells depend on the cell type, the origin of EVs, their content, and EVs can act locally and/or systemically. The changes in the recipient cells include modulation of the intracellular signaling pathways, gene regulation, post-transcriptional regulation, activation, or inhibition of different cell types [21, 22, 23]. After target cell recognition, EVs can interact with surface receptors, followed by fusion with the plasma membrane for releasing their content, and signaling different intracellular events. However, EVs can also be endocytosed by target cells or collapse after their secretion, delivering their contents into the intracellular space [8, 15].
In parasitic diseases, EVs have brought an exciting field to investigate since they can act as mediators in parasite–host interaction, allowing the transfer of virulence factors and effector molecules from the parasites to the host [24, 25, 26]. Parasites EVs are related to the pathogen adhesion, the spread of the parasites, and play a role in regulating the host’s immune system. In addition, immune cells infected and/or stimulated with parasite components can release EVs [23] containing messenger RNA (mRNA), small noncoding RNAs (microRNA), chromosomal and mitochondrial DNA, retrotransposons, parasites antigens, and major histocompatibility complex (MHC) I and II [23, 27]. The effects in immunity are diverse, including modulation of innate immune response and antigen presentation.
The production and releasing of EVs by parasites or parasitized cells have been described and characterized in several parasitic infections [25]. For example, in
3. EVs released by Leishmania spp
The release of EVs by
Proteomic studies showed the presence of the metalloprotease GP63 in EVs released by
Besides GP63, other proteins have already been identified in
The presence of small noncoding RNAs was identified in EVs released by
4. Leishmania -EVs and immune response
Some evidence have pointed that
Few studies have proposed mechanisms of intracellular signaling pathways activated by
Besides macrophages and DCs,
In a mammalian host,
Evidence suggests that EVs released by
5. EVs and leishmaniasis progression
Experimental models have contributed to better understanding the role of EVs in the leishmaniasis progression. The treatment of mice with
Changes in the content of EVs may impact the immune response and disease progression [9, 11]. Studies performed with genetically modified parasites showed that in a mouse model of air pouch formation (murine air pouch injection) EVs derived from
The relevance of EVs in
6. Conclusions
The knowledge acquired studying EVs has allowed understanding that these particles are related to intercellular communication and cross-kingdom relationship. The release of these EVs by
Biological function | Reference | |
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| [11] | |
| [5] | |
| [40] | |
| [45] | |
| [41] | |
| [42] | |
| [44] | |
| [10] |
Acknowledgments
This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (grant number 2019/21614-3). Scholarships were provided by the Fundação de Amparo à Pesquisa do Estado de São Paulo (2021/01556-9), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).
References
- 1.
Burza S, Croft SL, Boelaert M, Leishmaniasis. Lancet. 2018; 392 : 951-970. DOI: 10.1016/S0140-6736(18)31204-2 - 2.
Scott P, Novais FO, Cutaneous leishmaniasis: Immune responses in protection and pathogenesis. Nature Reviews Immunology. 2016; 16 (9):581-592. DOI: 10.1038/NRI.2016.72 - 3.
de Morais CG, Castro Lima AK, Terra R, dos Santos RF, Da-Silva SA, Dutra PM. The Dialogue of the Host-Parasite Relationship: Leishmania spp. andTrypanosoma cruzi Infection. BioMed Research International. 2015;2015 :324915. DOI: 10.1155/2015/324915 - 4.
Silverman JM, Clos J, de'Oliveira CC, Shirvani O, Fang Y, Wang C, et al. An exosome-based secretion pathway is responsible for protein export from Leishmania and communication with macrophages. Journal of Cell Science. 2010;123 :842-852. DOI: 10.1242/jcs.056465 - 5.
Barbosa FMC, Dupin TV, Toledo MDS, Reis NFDC, Ribeiro K, Cronemberger-Andrade A, et al. Extracellular vesicles released by Leishmania (Leishmania) amazonensis promote disease progression and induce the production of different cytokines in macrophages and B-1 cells. Frontiers in Microbiology. 2018;9 :3056. DOI: 10.3389/fmicb.2018.03056 - 6.
Atayde VD, Aslan H, Townsend S, Hassani K, Kamhawi S, Olivier M. Exosome secretion by the parasitic protozoan Leishmania within the sand fly midgut. Cell Reports. 2015;13 (5):957-967. DOI: 10.1016/j.celrep.2015.09.058 - 7.
Théry C, Witwer KW, Aikawa E, Alcaraz MJ, Anderson JD, Andriantsitohaina R, et al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. Journal of Extracellular Vesicles. 2018; 7 (1):1535750. DOI: 10.1080/20013078.2018.1535750 - 8.
van Niel G, D'Angelo G, Raposo G. Shedding light on the cell biology of extracellular vesicles. Nature Reviews Molecular Cell Biology. 2018; 19 (4):213-228. DOI: 10.1038/nrm.2017.125 - 9.
Hassani K, Shio MT, Martel C, Faubert D, Olivier M. Absence of metalloprotease GP63 alters the protein content of Leishmania exosomes. PLoS ONE. 2014;9 (4):e95007. DOI: 10.1371/journal.pone.0095007 - 10.
Atayde VD, Hassani K, da Silva Lira Filho A, Borges AR, Adhikari A, Martel C, et al. Leishmania exosomes and other virulence factors: Impact on innate immune response and macrophage functions. Cellular Immunology. 2016;309 :7-18. DOI: 10.1016/j.cellimm.2016.07.013 - 11.
Silverman JM, Clos J, Horakova E, Wang AY, Wiesgigl M, Kelly I, et al. Leishmania exosomes modulate innate and adaptive immune responses through effects on monocytes and dendritic cells. Journal of Immunology. 2010;185 (9):5011-5022. DOI: 10.4049/jimmunol.1000541 - 12.
Battistelli M, Falcieri E. Apoptotic bodies: Particular extracellular vesicles involved in intercellular communication. Biology (Basel). 2020; 9 (1):21. DOI: 10.3390/biology9010021 - 13.
Raposo G, Stoorvogel W. Extracellular vesicles: Exosomes, microvesicles, and friends. The Journal of Cell Biology. 2013; 200 :373-383. DOI: 10.1083/jcb.201211138 - 14.
Tkach M, Théry C. Communication by extracellular vesicles: Where we are and where we need to go. Cell 2016; 164 (6):1226-1232. DOI: 10.1016/j.cell.2016.01.043 - 15.
Meldolesi J. Exosomes and ectosomes in intercellular communication. Current Biology. 2018; 28 (8):R435-RR44. DOI: 10.1016/j.cub.2018.01.059 - 16.
Kowal J, Arras G, Colombo M, Jouve M, Morath JP, Primdal-Bengtson B, et al. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proceedings of the National Academy of Sciences of the United States of America. 2016; 113 (8):E968-E977. DOI: 10.1073/pnas.1521230113 - 17.
Cai H, Reinisch K, Ferro-Novick S. Coats, tethers, Rabs, and SNAREs work together to mediate the intracellular destination of a transport vesicle. Developmental Cell. 2007; 12 (5):671-682. DOI: 10.1016/j.devcel.2007.04.005 - 18.
Yáñez-Mó M, Siljander PR, Andreu Z, Zavec AB, Borràs FE, Buzas EI, et al. Biological properties of extracellular vesicles and their physiological functions. Journal of Extracellular Vesicles. 2015; 4 :27066. DOI: 10.3402/jev.v4.27066 - 19.
Gavinho B, Sabatke B, Feijoli V, Rossi IV, da Silva JM, Evans-Osses I, et al. Peptidylarginine deiminase inhibition abolishes the production of large extracellular vesicles from Giardia intestinalis , Affecting host-pathogen interactions by hindering adhesion to host cells, Frontiers in Cellular Infection Microbiology. 2020;10 :417. DOI: 10.3389/fcimb.2020.00417 - 20.
Schorey JS, Cheng Y, Singh PP, Smith VL. Exosomes and other extracellular vesicles in host-pathogen interactions. EMBO Reports. 2015; 16 (1):24-43. DOI: 10.15252/embr.201439363 - 21.
Campos JH, Soares RP, Ribeiro K, Andrade AC, Batista WL, Torrecilhas AC. Extracellular vesicles: Role in inflammatory responses and potential uses in vaccination in cancer and infectious diseases. Journal of Immunology Research. 2015; 2015 :832057. DOI: 10.1155/2015/832057 - 22.
Dong G, Filho AL, Olivier M. Modulation of host-pathogen communication by extracellular vesicles (EVs) of the protozoan parasite. Frontiers in Cellular and Infection Microbiology. 2019; 9 :100. DOI: 10.3389/fcimb.2019.00100 - 23.
Khosravi M, Mirsamadi ES, Mirjalali H, Zali MR. Isolation and functions of extracellular vesicles derived from parasites: The promise of a new era in immunotherapy, vaccination, and diagnosis. International Journal of Nanomedicine. 2020; 15 :2957-2969. DOI: 10.2147/IJN.S250993 - 24.
Montaner S, Galiano A, Trelis M, Martin-Jaular L, Del Portillo HA, Bernal D, et al. The role of extracellular vesicles in modulating the host immune response during parasitic infections. Frontiers in Immunology. 2014; 5 :433. DOI: 10.3389/fimmu.2014.00433 - 25.
Marcilla A, Martin-Jaular L, Trelis M, de Menezes-Neto A, Osuna A, Bernal D, et al. Extracellular vesicles in parasitic diseases. Journal of Extracelluar Vesicles. 2014; 3 :25040. DOI: 10.3402/jev.v3.25040 - 26.
Soares R, Xander P, Costa A, Marcilla A, Menezes-Neto A, Del Portillo H, et al. Highlights of the São Paulo ISEV workshop on extracellular vesicles in cross-kingdom communication. Journal of Extracellular Vesicles. 2017; 6 (1):1407213. DOI: 10.1080/20013078.2017.1407213 - 27.
Jeppesen DK, Fenix AM, Franklin JL, Higginbotham JN, Zhang Q, Zimmerman LJ, et al. Reassessment of exosome composition. Cell. 2019; 177 (2):428-445. DOI: 10.1016/j.cell.2019.02.029 - 28.
Lambertz U, Silverman JM, Nandan D, McMaster WR, Clos J, Foster LJ, et al. Secreted virulence factors and immune evasion in visceral leishmaniasis. Journal of Leukocyte Biology. 2012; 91 (6):887-899. DOI: 10.1189/jlb.0611326 - 29.
Silverman JM, Chan SK, Robinson DP, Dwyer DM, Nandan D, Foster LJ, et al. Proteomic analysis of the secretome of Leishmania donovani . Genome Biology. 2008;9 (2):R35. DOI: 10.1186/gb-2008-9-2-r35 - 30.
Hassani K, Antoniak E, Jardim A, Olivier M. Temperature-induced protein secretion by Leishmania mexicana modulates macrophage signalling and function. PLoS ONE. 2011;6 (5):e18724. DOI: 10.1371/journal.pone.0018724 - 31.
Gomez MA, Contreras I, Hallé M, Tremblay ML, McMaster RW, Olivier M. Leishmania GP63 alters host signaling through cleavage-activated protein tyrosine phosphatases. Sci Signal. 2009;2 (90):ra58. DOI: 10.1126/scisignal.2000213 - 32.
Hallé M, Gomez MA, Stuible M, Shimizu H, McMaster WR, Olivier M, et al. The Leishmania surface protease GP63 cleaves multiple intracellular proteins and actively participates in p38 mitogen-activated protein kinase inactivation. The Journal of Biological Chemistry. 2009;284 (11):6893-6908. DOI: 10.1074/jbc.M805861200 - 33.
Isnard A, Shio MT, Olivier M. Impact of Leishmania metalloprotease GP63 on macrophage signaling. Frontiers in Cellular and Infection Microbiology. 2012;2 :72. DOI: 10.3389/fcimb.2012.00072 - 34.
Olivier M, Atayde VD, Isnard A, Hassani K, Shio MT. Leishmania virulence factors: Focus on the metalloprotease GP63. Microbes and Infection. 2012;14 (15):1377-1389. DOI: 10.1016/j.micinf.2012.05.014 - 35.
Marshall S, Kelly PH, Singh BK, Pope RM, Kim P, Zhanbolat B, et al. Extracellular release of virulence factor major surface protease via exosomes in Leishmania infantum promastigotes. Parasites & Vectors. 2018;11 (1):355. DOI: 10.1186/s13071-018-2937-y - 36.
Santarém N, Racine G, Silvestre R, Cordeiro-da-Silva A, Ouellette M. Exoproteome dynamics in Leishmania infantum . Journal of Proteomics. 2013;84 :106-118. DOI: 10.1016/j.jprot.2013.03.012 - 37.
Forrest DM, Batista M, Marchini FK, Tempone AJ, Traub-Csekö YM. Proteomic analysis of exosomes derived from procyclic and metacyclic-like cultured Leishmania infantum chagasi . Journal of Proteomics. 2020;227 :103902. DOI: 10.1016/j.jprot.2020.103902 - 38.
Douanne N, Dong G, Douanne M, Olivier M, Fernandez-Prada C. Unravelling the proteomic signature of extracellular vesicles released by drug-resistant Leishmania infantum parasites. PLoS Neglected Tropical Diseases. 2020;14 (7):e0008439. DOI: 10.1371/journal.pntd.0008439 - 39.
Lambertz U, Oviedo Ovando ME, Vasconcelos EJ, Unrau PJ, Myler PJ, Reiner NE. Small RNAs derived from tRNAs and rRNAs are highly enriched in exosomes from both old and new world Leishmania providing evidence for conserved exosomal RNA Packaging. BMC Genomics. 2015;16 :151. DOI: 10.1186/s12864-015-1260-7 - 40.
Nogueira PM, de Menezes-Neto A, Borges VM, Descoteaux A, Torrecilhas AC, Xander P, et al. Immunomodulatory Properties of leishmania extracellular vesicles during host-parasite interaction: Differential activation of TLRs and NF-κB translocation by dermotropic and viscerotropic species. Frontiers in Cellular and Infection Microbiology. 2020;10 :380. DOI: 10.3389/fcimb.2020.00380 - 41.
Sauter IP, Madrid KG, de Assis JB, Sá-Nunes A, Torrecilhas AC, Staquicini DI, et al. TLR9/MyD88/TRIF signaling activates host immune inhibitory CD200 in Leishmania infection. JCI Insight. 2019;4 (10). DOI: 10.1172/jci.insight.126207 - 42.
de Carvalho RVH, Lima-Junior DS, da Silva MVG, Dilucca M, Rodrigues TS, Horta CV, et al. Leishmania RNA virus exacerbates Leishmaniasis by subverting innate immunity via TLR3-mediated NLRP3 inflammasome inhibition. Nature Communications. 2019;10 (1):5273. DOI: 10.1038/s41467-019-13356-2 - 43.
Olivier M, Zamboni DS. Leishmania Viannia guyanensis , LRV1 virus and extracellular vesicles: A dangerous trio influencing the faith of immune response during muco-cutaneous leishmaniasis. Current Opinion in Immunology. 2020;66 :108-113. DOI: 10.1016/j.coi.2020.08.004 - 44.
Belo R, Santarém N, Pereira C, Pérez-Cabezas B, Macedo F, Leite-de-Moraes M, et al. Exoproducts Inhibit Human Invariant NKT Cell Expansion and Activation. Frontiers in Immunology. 2017; 8 :710. DOI: 10.3389/fimmu.2017.00710 - 45.
Reis NFC, Dupin TV, Costa CR, Toledo MDS, de Oliveira VC, Popi AF, et al. Promastigotes or extracellular vesicles modulate B-1 cell activation and differentiation. Frontiers in Cellular and Infection Microbiology. 2020; 10 :573813. DOI: 10.3389/fcimb.2020.573813 - 46.
Hassani K, Olivier M. Immunomodulatory impact of leishmania -induced macrophage exosomes: A comparative proteomic and functional analysis. PLoS Neglected Tropical Diseases. 2013;7 (5):e2185. DOI: 10.1371/journal.pntd.0002185 - 47.
Cronemberger-Andrade A, Aragão-França L, de Araujo CF, Rocha VJ, Borges-Silva MaC, Figueira CP, et al. Extracellular vesicles from Leishmania -infected macrophages confer an anti-infection cytokine-production profile to naïve macrophages. PLoS Neglected Tropical Diseases. 2014;8 (9):e3161. DOI: 10.1371/journal.pntd.0003161 - 48.
Gioseffi A, Hamerly T, Van K, Zhang N, Dinglasan RR, Yates PA, et al. Leishmania-infected macrophages release extracellular vesicles that can promote lesion development. Life Science Alliance. 2020; 3 (12):e202000742. DOI: 10.26508/lsa.202000742 - 49.
Toledo MDS, Cronemberger-Andrade A, Barbosa FMC, Reis NFC, Dupin TV, Soares RP, et al. Effects of extracellular vesicles released by peritoneal B-1 cells on experimental Leishmania (Leishmania) amazonensis infection. Journal of Leukocyte Biology. 2020;108 (6):1803-1814. DOI: 10.1002/JLB.3MA0220-464RR