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J Extracell Vesicles
2015 Nov 26;4:28734. doi: 10.3402/jev.v4.28734.
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Vesicles from different Trypanosoma cruzi strains trigger differential innate and chronic immune responses.
Nogueira PM
,
Ribeiro K
,
Silveira AC
,
Campos JH
,
Martins-Filho OA
,
Bela SR
,
Campos MA
,
Pessoa NL
,
Colli W
,
Alves MJ
,
Soares RP
,
Torrecilhas AC
.
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Trypomastigote forms of Trypanosoma cruzi, the causative agent of Chagas Disease, shed extracellular vesicles (EVs) enriched with glycoproteins of the gp85/trans-sialidase (TS) superfamily and other α-galactosyl (α-Gal)-containing glycoconjugates, such as mucins. Here, purified vesicles from T. cruzi strains (Y, Colombiana, CL-14 and YuYu) were quantified according to size, intensity and concentration. Qualitative analysis revealed differences in their protein and α-galactosyl contents. Later, those polymorphisms were evaluated in the modulation of immune responses (innate and in the chronic phase) in C57BL/6 mice. EVs isolated from YuYu and CL-14 strains induced in macrophages higher levels of proinflammatory cytokines (TNF-α and IL-6) and nitric oxide via TLR2. In general, no differences were observed in MAPKs activation (p38, JNK and ERK 1/2) after EVs stimulation. In splenic cells derived from chronically infected mice, a different modulation pattern was observed, where Colombiana (followed by Y strain) EVs were more proinflammatory. This modulation was independent of the T. cruzi strain used in the mice infection. To test the functional importance of this modulation, the expression of intracellular cytokines after in vitro exposure was evaluated using EVs from YuYu and Colombiana strains. Both EVs induced cytokine production with the appearance of IL-10 in the chronically infected mice. A high frequency of IL-10 in CD4+ and CD8+ T lymphocytes was observed. A mixed profile of cytokine induction was observed in B cells with the production of TNF-α and IL-10. Finally, dendritic cells produced TNF-α after stimulation with EVs. Polymorphisms in the vesicles surface may be determinant in the immunopathologic events not only in the early steps of infection but also in the chronic phase.
Fig. 1. Procedures employed for the production, fractionation and characterization of T. cruzi EVs from different strains.
Fig. 2. Flow cytometry strategy for intracellular cytokine production by spleen cells from infected mice. (a) Dot plots of CD4, CD8, CD19, F4/80/CD11b and MHCII/CD11c expression. The representative dot plots illustrating the frequency (%) of cytokines from spleen cells are shown for Colombiana and YuYu strains. (b) CD4+IL-10+ cells. (c) CD8+IL-10+. (d) CD19+TNF-α+. (e) MHC/CD11c+TNF-α+.
Fig. 3.
Trypanosoma cruzi (Y strain) trypomastigotes spontaneously shed vesicles from their entire membrane surface. Scanning electron microscopy (SEM) of parasite membrane shedding after incubation in culture medium (a–d, bars: 1–5 µm). Magnification: (a) 27,383×, (b) 25,242×, (c) 60,470× and (d) 92,084×.
Fig. 4. Nanoparticle tracking analysis of the EVs isolated from different strains of T. cruzi. Graphic demonstration of size distribution and concentration for Y (a), Colombiana (b), CL-14 (c) and YuYu (d) strains. The average size (nm) (e) and concentration (particles/mL) (f) of the vesicles for all strains are represented. Data are representative of 3 independent experiments.
Fig. 6. Nitric oxide (NO) production by murine macrophages stimulated by EVs from CL-14 and YuYu strains is dependent on TLR2. Murine macrophages (C57BL/6, TLR2 − /− and TLR4 − /−) were stimulated with different concentrations of T. cruzi EVs (1, 5 and 50 µg). Cells were pre-incubated with IFN-γ (100 U/mL) for 18 h prior to addition of EVs or LPS, LPG and live parasites MOI 10:1 (positive controls). Negative controls included medium and medium + INF-γ. LPG Lb, L. braziliensis LPG; LPS, lipopolysaccharide (LPS) from E. coli; T. cruzi (Y), live parasites of T. cruzi (Y strain). Bars express the mean value±SD of 2 separate studies (*p < 0.05).
Fig. 7. TNF-α production by murine macrophages stimulated by EVs from CL-14 and YuYu strains is dependent on TLR2. Murine macrophages (C57BL/6, TLR2 − /− and TLR4 − /−) were stimulated with different concentrations of T. cruzi EVs (1, 5 and 50 µg). Cells were pre-incubated with IFN-γ (100 U/mL) for 18 h prior to addition of the EVs, and controls (LPS, LPG and live parasites MOI 10:1) (positive controls). Negative controls included medium and medium + INF-γ. IFN-γ, gamma-interferon; LPG Lb, L. braziliensis LPG; LPS, lipopolysaccharide from E. coli; T. cruzi (Y), live parasites of T. cruzi (Y strain). Bars express the mean value±SD of 2 separate studies (*p < 0.05).
Fig. 8.
Trypanosoma cruzi EVs equally activate MAPKs (ERK 1/2, p38 and JNK) from J774.1 macrophages. Cells were stimulated with EVs (5 µg/mL) at different time points (5, 15, 30 and 45 min). Dually phosphorylated MAPKs were detected by western blot: (a) ERK 1/2, (b) p38 and (c) JNK. C-, negative control (medium); C+, positive control (LPS from E. coli) (100 ng/mL).
Acosta-Serrano,
The mucin-like glycoprotein super-family of Trypanosoma cruzi: structure and biological roles.
2001, Pubmed
Acosta-Serrano,
The mucin-like glycoprotein super-family of Trypanosoma cruzi: structure and biological roles.
2001,
Pubmed
Almeida,
Highly purified glycosylphosphatidylinositols from Trypanosoma cruzi are potent proinflammatory agents.
2000,
Pubmed
Almeida,
Lytic anti-alpha-galactosyl antibodies from patients with chronic Chagas' disease recognize novel O-linked oligosaccharides on mucin-like glycosyl-phosphatidylinositol-anchored glycoproteins of Trypanosoma cruzi.
1994,
Pubmed
Almeida,
Complement-mediated lysis of Trypanosoma cruzi trypomastigotes by human anti-alpha-galactosyl antibodies.
1991,
Pubmed
Alves,
Trypanosoma cruzi: adhesion to the host cell and intracellular survival.
2007,
Pubmed
Andrews,
Adhesion and interiorization of Trypanosoma cruzi in mammalian cells.
1982,
Pubmed
Arthur,
Mitogen-activated protein kinases in innate immunity.
2013,
Pubmed
Assis,
Glycoinositolphospholipids from Leishmania braziliensis and L. infantum: modulation of innate immune system and variations in carbohydrate structure.
2012,
Pubmed
Basso,
Experimental Chagas disease. Innate immune response in Balb/c mice previously vaccinated with Trypanosoma rangeli. I. The macrophage shows immunological memory: Reality or fiction?
2014,
Pubmed
Basso,
Modulation of immune response in experimental Chagas disease.
2013,
Pubmed
Bermejo,
BAFF mediates splenic B cell response and antibody production in experimental Chagas disease.
2010,
Pubmed
Bryan,
Specific humoral immunity versus polyclonal B cell activation in Trypanosoma cruzi infection of susceptible and resistant mice.
2010,
Pubmed
Burleigh,
Host cell signaling and Trypanosoma cruzi invasion: do all roads lead to lysosomes?
2005,
Pubmed
Burleigh,
Cell signalling and Trypanosoma cruzi invasion.
2002,
Pubmed
Campos,
Activation of Toll-like receptor-2 by glycosylphosphatidylinositol anchors from a protozoan parasite.
2001,
Pubmed
Campos,
Extracellular Vesicles: Role in Inflammatory Responses and Potential Uses in Vaccination in Cancer and Infectious Diseases.
2015,
Pubmed
,
Echinobase
Caradonna,
Mechanisms of host cell invasion by Trypanosoma cruzi.
2011,
Pubmed
Costa,
Functional IL-10 gene polymorphism is associated with Chagas disease cardiomyopathy.
2009,
Pubmed
Coura,
Chagas disease: a new worldwide challenge.
2010,
Pubmed
Dc-Rubin,
T rypanosoma cruzi trans-sialidase as a multifunctional enzyme in Chagas' disease.
2012,
Pubmed
Di Girolamo,
Plasmodium lipid rafts contain proteins implicated in vesicular trafficking and signalling as well as members of the PIR superfamily, potentially implicated in host immune system interactions.
2008,
Pubmed
Duddy,
Distinct profiles of human B cell effector cytokines: a role in immune regulation?
2004,
Pubmed
Dutra,
Self and nonself stimulatory molecules induce preferential expansion of CD5+ B cells or activated T cells of chagasic patients, respectively.
2000,
Pubmed
Dutra,
Immunoregulatory networks in human Chagas disease.
2014,
Pubmed
Dutra,
Activated T and B lymphocytes in peripheral blood of patients with Chagas' disease.
1994,
Pubmed
Fares,
Identification of phenotypic markers of B cells from patients with Chagas disease.
2013,
Pubmed
Ferguson,
The structure, biosynthesis and functions of glycosylphosphatidylinositol anchors, and the contributions of trypanosome research.
1999,
Pubmed
Galili,
The alpha-gal epitope and the anti-Gal antibody in xenotransplantation and in cancer immunotherapy.
2005,
Pubmed
Gazzinelli,
Direct lysis of Trypanosoma cruzi: a novel effector mechanism of protection mediated by human anti-gal antibodies.
1991,
Pubmed
Gazzinelli,
Role of the Toll/interleukin-1 receptor signaling pathway in host resistance and pathogenesis during infection with protozoan parasites.
2004,
Pubmed
Gazzinelli,
Protozoan encounters with Toll-like receptor signalling pathways: implications for host parasitism.
2006,
Pubmed
Geiger,
Exocytosis and protein secretion in Trypanosoma.
2010,
Pubmed
Giordanengo,
Cruzipain, a major Trypanosoma cruzi antigen, conditions the host immune response in favor of parasite.
2002,
Pubmed
Gonçalves,
Trypanosoma cruzi: shedding of surface antigens as membrane vesicles.
1991,
Pubmed
Gravina,
Differential use of TLR2 and TLR9 in the regulation of immune responses during the infection with Trypanosoma cruzi.
2013,
Pubmed
Hassani,
Immunomodulatory impact of leishmania-induced macrophage exosomes: a comparative proteomic and functional analysis.
2013,
Pubmed
Hong,
An important role of α-hemolysin in extracellular vesicles on the development of atopic dermatitis induced by Staphylococcus aureus.
2014,
Pubmed
Ibraim,
Two biochemically distinct lipophosphoglycans from Leishmania braziliensis and Leishmania infantum trigger different innate immune responses in murine macrophages.
2013,
Pubmed
Lima,
Negative tissue parasitism in mice injected with a noninfective clone of Trypanosoma cruzi.
1995,
Pubmed
Magdesian,
A conserved domain of the gp85/trans-sialidase family activates host cell extracellular signal-regulated kinase and facilitates Trypanosoma cruzi infection.
2007,
Pubmed
Marcilla,
Extracellular vesicles in parasitic diseases.
2014,
Pubmed
Marcilla,
Extracellular vesicles from parasitic helminths contain specific excretory/secretory proteins and are internalized in intestinal host cells.
2012,
Pubmed
Mattos,
The Gp85 surface glycoproteins from Trypanosoma cruzi.
2014,
Pubmed
Mott,
Modulation of host cell mechanics by Trypanosoma cruzi.
2009,
Pubmed
Oliveira,
Expression of functional TLR4 confers proinflammatory responsiveness to Trypanosoma cruzi glycoinositolphospholipids and higher resistance to infection with T. cruzi.
2004,
Pubmed
Ouaissi,
Molecular basis of Trypanosoma cruzi and Leishmania interaction with their host(s): exploitation of immune and defense mechanisms by the parasite leading to persistence and chronicity, features reminiscent of immune system evasion strategies in cancer diseases.
2005,
Pubmed
Paranaíba,
Leishmania enriettii: biochemical characterisation of lipophosphoglycans (LPGs) and glycoinositolphospholipids (GIPLs) and infectivity to Cavia porcellus.
2015,
Pubmed
,
Echinobase
Pineda,
Interactions of human galectins with Trypanosoma cruzi: binding profile correlate with genetic clustering of lineages.
2015,
Pubmed
Rassi,
Chagas disease.
2010,
Pubmed
Ropert,
Requirement of mitogen-activated protein kinases and I kappa B phosphorylation for induction of proinflammatory cytokines synthesis by macrophages indicates functional similarity of receptors triggered by glycosylphosphatidylinositol anchors from parasitic protozoa and bacterial lipopolysaccharide.
2001,
Pubmed
Ropert,
Regulatory role of Toll-like receptor 2 during infection with Trypanosoma cruzi.
2004,
Pubmed
Sathler-Avelar,
Benznidazole treatment during early-indeterminate Chagas' disease shifted the cytokine expression by innate and adaptive immunity cells toward a type 1-modulated immune profile.
2006,
Pubmed
Sathler-Avelar,
Blood leukocytes from benznidazole-treated indeterminate chagas disease patients display an overall type-1-modulated cytokine profile upon short-term in vitro stimulation with Trypanosoma cruzi antigens.
2012,
Pubmed
Sathler-Avelar,
Innate immunity and regulatory T-cells in human Chagas disease: what must be understood?
2009,
Pubmed
Silverman,
Leishmania exosomes deliver preemptive strikes to create an environment permissive for early infection.
2011,
Pubmed
Silverman,
Leishmania exosomes modulate innate and adaptive immune responses through effects on monocytes and dendritic cells.
2010,
Pubmed
Silverman,
An exosome-based secretion pathway is responsible for protein export from Leishmania and communication with macrophages.
2010,
Pubmed
Soares,
Intraspecies variation in Trypanosoma cruzi GPI-mucins: biological activities and differential expression of α-galactosyl residues.
2012,
Pubmed
Soares,
Leishmania braziliensis: a novel mechanism in the lipophosphoglycan regulation during metacyclogenesis.
2005,
Pubmed
Souza,
Trypanosoma cruzi infection induces differential modulation of costimulatory molecules and cytokines by monocytes and T cells from patients with indeterminate and cardiac Chagas' disease.
2007,
Pubmed
Souza,
Monocytes from patients with indeterminate and cardiac forms of Chagas' disease display distinct phenotypic and functional characteristics associated with morbidity.
2004,
Pubmed
Talvani,
Kinetics of cytokine gene expression in experimental chagasic cardiomyopathy: tissue parasitism and endogenous IFN-gamma as important determinants of chemokine mRNA expression during infection with Trypanosoma cruzi.
2000,
Pubmed
Torrecilhas,
Vesicles as carriers of virulence factors in parasitic protozoan diseases.
2012,
Pubmed
,
Echinobase
Torrecilhas,
Interference of natural mouse hepatitis virus infection with cytokine production and susceptibility to Trypanosoma cruzi.
1999,
Pubmed
Trocoli Torrecilhas,
Trypanosoma cruzi: parasite shed vesicles increase heart parasitism and generate an intense inflammatory response.
2009,
Pubmed
Tyler,
The life cycle of Trypanosoma cruzi revisited.
2001,
Pubmed
Uphoff,
Detection of mycoplasma contaminations.
2005,
Pubmed
Vitelli-Avelar,
Strategy to assess the overall cytokine profile of circulating leukocytes and its association with distinct clinical forms of human Chagas disease.
2008,
Pubmed
Zingales,
A new consensus for Trypanosoma cruzi intraspecific nomenclature: second revision meeting recommends TcI to TcVI.
2009,
Pubmed
