147 results on '"José Roberto Mineo"'
Search Results
2. Association of gestational diabetes mellitus and negative modulation of the specific humoral and cellular immune response against Toxoplasma gondii
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Ana Carolina de Morais Oliveira-Scussel, Paula Tatiana Mutão Ferreira, Renata de Souza Resende, Cristhianne Molinero Ratkevicius-Andrade, Angelica de Oliveira Gomes, Marina Carvalho Paschoini, Fernanda Bernadelli De Vito, Thaís Soares Farnesi-de-Assunção, Marcos Vinícius da Silva, José Roberto Mineo, Denise Bertulucci Rocha Rodrigues, and Virmondes Rodrigues
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Immunology ,Immunology and Allergy - Abstract
In order to evaluate and compare the specific immune response of pregnant women (PW) chronically infected withToxoplasma gondii, with and without gestational diabetes mellitus (GDM), and the humoral response of their respective newborns (NB), the study was carried out on 81 PW (34 GDM and 47 controls) from whose medical records the results of the oral glucose tolerance test (OGTT) were obtained, and blood samples were collected at the third trimester of pregnancy; also, on 45 NBs (20 GDM and 25 controls) from whom umbilical cord blood samples were obtained. Humoral immunity was analyzed by measuring anti-T. gondiitotal IgG, IgG subclasses and IgG avidity. To evaluate cellular immunity, peripheral blood mononuclear cells (PBMC) from 32 PW (16 GDM and 16 controls) were cultured, supernatant cytokines were determined, and flow cytometry was performed to analyze the expression at lymphocytes of surface molecules, cytokines and transcription factors. All PW and NBs were positive for total IgG, and the prevalent subclass was IgG1. There was a negative correlation between the OGTT glycemia of PW and the levels of total IgG, IgG1 and IgG avidity. The IgG avidity of the GDM group was significantly lower than the control group. Patients from the GDM group had a higher number of T lymphocytes expressing markers of cell activation and exhaustion (CD28 and PD-1). In the presence ofT. gondiisoluble antigen (STAg) the amount of CD4+T cells producing IFN-γ, IL-10 and IL-17 was significantly lower in the GDM group, while there was no difference between groups in the number of CD4+CD25HighFOXP3+LAP+ functional Treg cells. Additionally, under STAg stimulus, the secretion of IL-17, IL-4, TNF and IL-2 cytokines at PBMCs culture supernatant was lower in the GDM group. In conclusion, there was a correlation between the increase in blood glucose and the decrease in levels of anti-T. gondiiantibodies, associated with the decreased IgG avidity in patients who develop GDM. Also, the GDM group had decreased immune responses in Th1, Th2 and Th17 profiles, suggesting an association between GDM and the negative modulation of the humoral and cellular immune responses againstT. gondii.
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- 2022
3. Macrophage Migration Inhibitory Factor contributes to drive phenotypic and functional macrophages activation in response to Toxoplasma gondii infection
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Paula Tatiane Mutão Ferreira, Ana Carolina Morais Oliveira-Scussel, Roberto Augusto Pereira Sousa, Beatriz Quaresemin Gomes, Jhennifer Estevão Félix, Rafaela José Silva, Iliana Balga Millian, Thais Soares Farnesi Assunção, Samuel Cota Teixeira, Marcos de Lucca Moreira Gomes, Marcos Vinícius Silva, Bellisa Freitas Barbosa, Virmondes Rodrigues Junior, José Roberto Mineo, Carlo José Freire Oliveira, Eloisa Amália Vieira Ferro, and Angelica Oliveira Gomes
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Immunology ,Immunology and Allergy ,Hematology - Published
- 2023
4. Comparative Detection of Immunoglobulin Isotypes and Subclasses against
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Hellen Dayane Silva, Borges, Ana Carolina Morais, Oliveira-Scussel, Ângela Maria Morais, Oliveira, Vânia Olivetti Steffen, Abdallah, Ana Cláudia Arantes Marquez, Pajuaba, and José Roberto, Mineo
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Immunoglobulin Isotypes ,Immunoglobulin M ,Pregnancy ,Colostrum ,Immunoglobulin G ,Infant, Newborn ,Antibodies, Protozoan ,Humans ,Enzyme-Linked Immunosorbent Assay ,Female ,Toxoplasma ,Immunoglobulin A - Abstract
The study included 283 puerperal patients. ELISA (Enzyme-Linked Immunosorbent Assay) for detection of anti-It was found that 45.9%, 6.0%, and 2.1% of serum samples and 45.2%, 7.1%, and 2.1% of colostrum samples were positive for IgG, IgM, and IgA, respectively. Specific IgG1, IgG3, and IgG4 were positive, respectively, in 98.5%, 54.6%, and 44.6% of serum samples, in contrast with 56.9%, 78.5%, and 34.6% of colostrum samples. Thus, the predominant reactivity of IgG subclasses againstColostrum presents representative levels of IgM, IgA, IgG1, IgG3, and IgG4 antibodies specific to
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- 2022
5. Association of gestational diabetes mellitus and negative modulation of the specific humoral and cellular immune response against
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Ana Carolina de Morais, Oliveira-Scussel, Paula Tatiana Mutão, Ferreira, Renata de Souza, Resende, Cristhianne Molinero, Ratkevicius-Andrade, Angelica de Oliveira, Gomes, Marina Carvalho, Paschoini, Fernanda Bernadelli, De Vito, Thaís Soares, Farnesi-de-Assunção, Marcos Vinícius, da Silva, José Roberto, Mineo, Denise Bertulucci Rocha, Rodrigues, and Virmondes, Rodrigues
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Blood Glucose ,Immunity, Cellular ,Interleukin-17 ,Programmed Cell Death 1 Receptor ,Infant, Newborn ,Antibodies, Protozoan ,Interleukin-10 ,Diabetes, Gestational ,CD28 Antigens ,Pregnancy ,Immunoglobulin G ,Leukocytes, Mononuclear ,Cytokines ,Humans ,Interleukin-2 ,Female ,Interleukin-4 ,Toxoplasma ,Transcription Factors - Abstract
In order to evaluate and compare the specific immune response of pregnant women (PW) chronically infected with
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- 2022
6. LPS-mediated activation of TLR4 controls Toxoplasma gondii growth in human trophoblast cell (BeWo) and human villous explants in a dependent-manner of TRIF, MyD88, NF-κB and cytokines
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Alessandra Monteiro Rosini, Samuel Cota Teixeira, Iliana Claudia Balga Milian, Rafaela José Silva, Guilherme de Souza, Luana Carvalho Luz, Angelica Oliveira Gomes, José Roberto Mineo, Tiago Wilson Patriarca Mineo, Eloisa Amália Vieira Ferro, and Bellisa Freitas Barbosa
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Lipopolysaccharides ,NF-kappa B ,Cell Biology ,General Medicine ,Interleukin-10 ,Trophoblasts ,Toll-Like Receptor 4 ,Transforming Growth Factor beta1 ,Adaptor Proteins, Vesicular Transport ,Cell Line, Tumor ,Myeloid Differentiation Factor 88 ,Cytokines ,Humans ,Toxoplasma ,Developmental Biology ,Adaptor Proteins, Signal Transducing - Abstract
We evaluated the influence of the Toll-like receptor (TLR)-4 pathways on BeWo, JEG-3 and HTR-8/SVneo cells, as well as in human villous explants infected with Toxoplasma gondii. Cells and explants were stimulated with LPS for 24 or 48 h and processed for the MTT assay, and expression of TLR4 was evaluated by confocal microscopy. In addition, we used peptides that inhibit MyD88 or TRIF, and inhibitor to NF-κB. Finally, the parasite proliferation was verified, and ELISA was performed to verify the cytokine production. As results, LPS did not induce toxicity in cells and explants. However, LPS triggered a reduction in T. gondii proliferation only in BeWo cells and explants. Additionally, LPS downmodulated IL-10, TGF-β1 and TNF, but upregulated IFN-γ in BeWo cells. For explants, LPS induced high levels of IL-10, TGF-β1 and IFN-γ. Finally, it was observed that the inhibition of TRIF and NF-κB increased parasitism and modulated TGF-β1 in BeWo cells, while the inhibition of MyD88 and NF-κB increased T. gondii infection and modulated IFN-γ in explants. It can be concluded that the TLR4 pathway is important for the control of T. gondii replication in BeWo cells and villous explants, in a dependent-manner of TRIF, MyD88, NF-κB and cytokines.
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- 2022
7. Role of TLR2/MyD88 in the production of specific IgM and IgG antibodies during the immunization of mice against Neospora caninum
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Mariana Ferreira Silva, Carolina Salomão Lopes, Flávia Batista Ferreira França, Eliézer Lucas Pires Ramos, Fernanda Maria Santiago, José Roberto Mineo, and Tiago Wilson Patriarca Mineo
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General Veterinary ,General Immunology and Microbiology ,Coccidiosis ,Public Health, Environmental and Occupational Health ,Neospora ,Nitric Oxide Synthase Type II ,Interleukin-12 ,Toll-Like Receptor 2 ,Interferon-gamma ,Mice ,Infectious Diseases ,Immunoglobulin M ,Immunoglobulin G ,Myeloid Differentiation Factor 88 ,Molecular Medicine ,Animals ,Immunization - Abstract
Neospora caninum is a parasite relevant to the veterinary field. Innate and adaptive responses against N. caninum induce effector mechanisms that limit parasite replication, but little is known about their role in humoral response. Our work aimed to verify whether key molecules in the TLR2/MyD88-mediated response would impact the production of specific IgM and IgG antibodies in mice during immunization with soluble antigens of N. caninum. We observed that lack of IFN-gamma did not negatively affect the production of specific antibodies. However, mice genetically deficient in Toll-like receptor 2, Myeloid differentiation factor 88, Interleukin 12 and inducible nitric oxide synthase presented significant decrease in antibody levels against N. caninum antigens, which also reflected in the diversity of the antigen recognized by their serum. In that sense, we show here that molecules within this innate recognition pathway may present a direct impact in the induction of an antibody response against N. caninum.
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- 2022
8. Altered visual attention behavior of Toxoplasma gondii-infected individuals
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Fernanda P. Dirscherl, José Roberto Mineo, Deise A. O. Silva, Carolina Salomão Lopes, and Joaquim Carlos Rossini
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Neuropsychology and Physiological Psychology ,biology ,General Neuroscience ,Immunology ,Toxoplasma gondii ,Visual attention ,biology.organism_classification ,Psychology - Published
- 2019
9. Editorial: The Effects of Physical Activity and Exercise on Immune Responses to Infection
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Patricia M. L. Dutra, Silvia A. G. Da-Silva, José Roberto Mineo, and James E. Turner
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Immunology ,Immunity ,physical activity ,Immunology and Allergy ,Humans ,infections ,Immunologic diseases. Allergy ,RC581-607 ,Exercise ,immune response ,exercise immunology - Published
- 2021
10. Sulfadiazine Plus Pyrimethamine Therapy Reversed Multiple Behavioral and Neurocognitive Changes in Long-Term Chronic Toxoplasmosis by Reducing Brain Cyst Load and Inflammation-Related Alterations
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Barrios Leda Castaño, Andrea Alice Silva, Lina L Hernandez-Velasco, Ana Paula Da Silva Pinheiro, Daniel Gibaldi, José Roberto Mineo, Neide Maria Silva, and Joseli Lannes-Vieira
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Inflammation ,Mice, Inbred C57BL ,Memory Disorders ,Mice ,Pyrimethamine ,Immunology ,Immunology and Allergy ,Animals ,Brain ,Cytokines ,Sulfadiazine ,Female ,Toxoplasmosis - Abstract
Toxoplasma gondiiinfects one-third of the world population. For decades, it has been considered a silent lifelong infection. However, chronicallyT. gondii-infected persons may present psychiatric and neurocognitive changes as anxiety, depression, and memory loss. In a model of long-term chronic infection, behavioral alterations parallel neuroinflammation and systemic high cytokine levels, and may reflect brain cyst load. Recent findings support that in chronic infection an active parasite-host interplay involves an immune-mediated control of tissue cysts. Here, we tested the idea that etiological treatment in chronic phase may add advantage to intrinsic immune-mediated cyst control and impact behavioral changes. Thus, we combined sulfadiazine-plus-pyrimethamine (S+P), the first-choice therapy for toxoplasmosis, to study the association of brain cyst load and biological processes related to the immune response (neuroinflammation, blood-brain barrier -BBB- disruption and serum cytokine levels), with behavioral and neurocognitive changes of long-term chronic infection. Female C57BL/6 mice (H-2b) were infected (5 cysts, ME-49 strain) and treated with S+P from 30 to 60 days postinfection (dpi), compared with vehicle (Veh)-treated and noninfected controls. At endpoints (pre-therapy, 30 dpi; S+P therapy, 60 dpi; after ceased therapy, 90 dpi), independent groups were subjected to behavioral tests, and brain tissues and sera were collected. Multiple behavioral and neurocognitive changes were detected in the early (30 dpi) and long-term (60 and 90 dpi) chronic infection. S+P therapy resolved locomotor alterations, anxiety, and depressive-like behavior, partially or transiently ameliorated hyperactivity and habituation memory loss. Analysis after therapy cessation showed that S+P therapy reduced the number of stimuli required for aversive memory consolidation. S+P therapy resulted in reduced brain cyst load, neuroinflammation and BBB disruption, and lowered systemic Th1-cytokine levels. Correlation analysis revealed association between IFNγ, TNF and MCP-1/CCL2 serum levels, brain cyst load and behavioral and neurocognitive alterations. Moreover, principal-component analysis (PCA-2D and 3D projections) highlighted distinction between clusters (noninfected; Veh-treated and S+P-treated infected). Thus, our data suggest that S+P therapy added gain to intrinsic brain cyst control and, direct or indirectly, ameliorated inflammation-related alterations, traits associated with behavioral and neurocognitive alterations.
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- 2021
11. TNF-TNFR1 Signaling Enhances the Protection Against
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Flávia Batista, Ferreira França, Murilo Vieira, Silva, Mariana Ferreira, Silva, Eliézer Lucas Pires, Ramos, Vanessa Dos Santos, Miranda, Caroline Martins, Mota, Fernanda Maria, Santiago, José Roberto, Mineo, and Tiago Wilson Patriarca, Mineo
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Mice, Knockout ,Coccidiosis ,Tumor Necrosis Factor-alpha ,Neospora ,TNF ,respiratory system ,effector molecules ,Mice, Inbred C57BL ,Mice ,Cellular and Infection Microbiology ,Pregnancy ,Receptors, Tumor Necrosis Factor, Type I ,parasitic diseases ,Animals ,Cytokines ,antibodies ,Female ,neosporosis ,chronic phase ,Original Research - Abstract
Neospora caninum is a protozoan associated with abortions in ruminants and neuromuscular disease in dogs. Classically, the immune response against apicomplexan parasites is characterized by the production of proinflammatory cytokines, such as IL-12, IFN-γ and TNF. TNF is mainly produced during the acute phases of the infections and binds to TNF receptor 1 (CD120a, p55, TNFR1) activating a variety of cells, hence playing an important role in the induction of the inflammatory process against diverse pathogens. Thus, in this study, we aimed to evaluate the role of TNF in cellular and humoral immune responses during N. caninum infection. For this purpose, we used a mouse model of infection based on wildtype (WT) and genetically deficient C57BL/6 mice in TNFR1 (Tnfr1 -/-). We observed that Tnfr1 -/- mice presented higher mortality associated with inflammatory lesions and increased parasite burden in the brain after the infection with N. caninum tachyzoites. Moreover, Tnfr1 -/- mice showed a reduction in nitric oxide (NO) levels in vivo. We also observed that Tnfr1 -/- mice showed enhanced serum concentration of antigen-specific IgG2 subclass, while IgG1 production was significantly reduced compared to WT mice, suggesting that TNFR1 is required for regular IgG subclass production and antigen recognition. Based on our results, we conclude that the TNF-TNFR1 complex is crucial for mediating host resistance during the infection by N. caninum.
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- 2021
12. Comparative Detection of Immunoglobulin Isotypes and Subclasses against Toxoplasma gondii Soluble Antigen in Serum and Colostrum Samples from Puerperal Women
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Hellen Dayane Silva Borges, Ana Carolina Morais Oliveira-Scussel, Ângela Maria Morais Oliveira, Vânia Olivetti Steffen Abdallah, Ana Cláudia Arantes Marquez Pajuaba, and José Roberto Mineo
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Toxoplasma gondii ,colostrum ,serum ,antibody isotypes ,Health, Toxicology and Mutagenesis ,Public Health, Environmental and Occupational Health - Abstract
Background: Toxoplasma gondii is an obligate intracellular parasite that can infect several species, including humans, and can cause severe damage to the fetus when the infection occurs during pregnancy. The environment and/or food contamination are critical to spreading the infection. Human milk is rich in nutrients and bioactive elements that provide growth and development of the immune system of the newborn. All isotypes of immunoglobulins are present in human colostrum and they are produced from systemic or local sources. Breastfeeding protects the infant against various pathogens, but there is no conclusive study to detect IgG subclasses in colostrum against T. gondii. Therefore, the aim of this study was to detect and evaluate the presence of antibody isotypes against T. gondii in paired samples of serum and colostrum. Methods: The study included 283 puerperal patients. ELISA (Enzyme-Linked Immunosorbent Assay) for detection of anti-T. gondii-specific IgM, IgA, and IgG isotypes and IgG1, IgG3, and IgG4 subclasses were conducted on paired samples of serum and colostrum. Results: It was found that 45.9%, 6.0%, and 2.1% of serum samples and 45.2%, 7.1%, and 2.1% of colostrum samples were positive for IgG, IgM, and IgA, respectively. Specific IgG1, IgG3, and IgG4 were positive, respectively, in 98.5%, 54.6%, and 44.6% of serum samples, in contrast with 56.9%, 78.5%, and 34.6% of colostrum samples. Thus, the predominant reactivity of IgG subclasses against T. gondii was IgG1 in serum and IgG3 in colostrum. The higher percentage of positive samples and higher levels of anti-T. gondii IgG3 antibodies were observed in colostrum, when compared to serum samples, suggesting a local production of this subclass. IgG3 and IgG1 subclasses presented different percentages of positivity in serum and colostrum. Only the IgG1 subclass showed a significant correlation between the levels of anti-T. gondii in serum and colostrum, suggesting that IgG1 in breast milk comes from a systemic source. IgG4 showed a similar percentage of positivity in both sample types, but no significant correlation was observed between their levels. Conclusion: Colostrum presents representative levels of IgM, IgA, IgG1, IgG3, and IgG4 antibodies specific to T. gondii. The detection of these antibodies presents the potential for diagnostic application of colostrum samples to better identify the diagnostic status of T. gondii infection, especially during the acute phase. In addition, breastfeeding can also be a possible source of protective antibodies for the newborn against toxoplasmosis, an anthropozoonosis maintained by environmental infection, which interferes in the public health of many countries.
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- 2022
13. Transforming growth factor (TGF)-β1 and interferon (IFN)-γ differentially regulate ICAM-1 expression and adhesion of Toxoplasma gondii to human trophoblast (BeWo) and uterine cervical (HeLa) cells
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Samuel Cota Teixeira, B.F. Barbosa, Eloisa Amália Vieira Ferro, Rafaela José da Silva, M.B. Angeloni, Deise A. O. Silva, Marise Lopes Fermino, A.O. Gomes, Janice Buiate Lopes-Maria, Maria Cristina Roque-Barreira, José Roberto Mineo, and Neide M. Silva
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Veterinary (miscellaneous) ,medicine.medical_treatment ,HeLa ,Transforming Growth Factor beta1 ,Interferon ,parasitic diseases ,medicine ,Humans ,ICAM-1 ,biology ,Trophoblast ,Toxoplasma gondii ,Intercellular adhesion molecule ,biology.organism_classification ,Intercellular Adhesion Molecule-1 ,Molecular biology ,Trophoblasts ,Infectious Diseases ,medicine.anatomical_structure ,Cytokine ,Insect Science ,embryonic structures ,Parasitology ,Interferons ,Toxoplasma ,Transforming growth factor ,medicine.drug ,HeLa Cells - Abstract
Toxoplasma gondii is a parasite able to infect various cell types, including trophoblast cells. Studies have demonstrated that interleukin (IL)-10, transforming growth factor (TGF)-β1 and interferon (IFN)-γ are involved in the susceptibility of BeWo trophoblast cells to T. gondii infection. Furthermore, T. gondii is able to adhere to the plasma membrane of host cells through intercellular adhesion molecule (ICAM)-1. Thus, the present study aimed to assess the role of IL-10, TGF-β1 and IFN-γ in the expression of ICAM-1 in BeWo and HeLa cells and to analyze the role of ICAM-1 in the adhesion and invasion of T. gondii to these cells under the influence of these cytokines. For this purpose, BeWo and HeLa cells were treated or not, before and after T. gondii infection, with rIL-10, rTGF-β1 or rIFN-γ. For the BeWo cells, rIL-10 and rTGF-β1 favored susceptibility to infection, but only rTGF-β1 and rIFN-γ increased ICAM-1 expression, and TNF-α release. On the other hand, rIFN-γ downregulated the expression of ICAM-1 triggered by T. gondii in HeLa cells, leading to control of the infection. Moreover, we observed that upregulation of ICAM-1, mediated by cytokine's stimulation, in BeWo and HeLa cells resulted in a high number rate of both parasite adhesion and invasion to these cells, which were strongly reduced after ICAM-1 neutralization. Likewise, the blockage of ICAM-1 molecule also impaired T. gondii infection in human villous explants. Taken together, these findings demonstrate that TGF-β1 and IFN-γ differentially regulate ICAM-1 expression, which may interfere in the adhesion/invasion of T. gondii to BeWo and HeLa cells for modulating susceptibility to infection.
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- 2021
14. Biogenic Silver Nanoparticles Can Control
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Idessania Nazareth, Costa, Mayara, Ribeiro, Priscila, Silva Franco, Rafaela José, da Silva, Thádia Evelyn, de Araújo, Iliana Claudia Balga, Milián, Luana Carvalho, Luz, Pâmela Mendonça, Guirelli, Gerson, Nakazato, José Roberto, Mineo, Tiago W P, Mineo, Bellisa Freitas, Barbosa, and Eloisa Amália Vieira, Ferro
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nanoparticles treatment ,placenta ,congenital toxoplasmosis ,embryonic structures ,Toxoplasma gondii ,Microbiology ,trophoblast ,Original Research - Abstract
The combination of sulfadiazine and pyrimethamine plus folinic acid is the conventional treatment for congenital toxoplasmosis. However, this classical treatment presents teratogenic effects and bone marrow suppression. In this sense, new therapeutic strategies are necessary to reduce these effects and improve the control of infection. In this context, biogenic silver nanoparticles (AgNp-Bio) appear as a promising alternative since they have antimicrobial, antiviral, and antiparasitic activity. The purpose of this study to investigate the action of AgNp-Bio in BeWo cells, HTR-8/SVneo cells and villous explants and its effects against Toxoplasma gondii infection. Both cells and villous explants were treated with different concentrations of AgNp-Bio or combination of sulfadiazine + pyrimethamine (SDZ + PYZ) in order to verify the viability. After, cells and villi were infected and treated with AgNp-Bio or SDZ + PYZ in different concentrations to ascertain the parasite proliferation and cytokine production profile. AgNp-Bio treatment did not reduce the cell viability and villous explants. Significant reduction was observed in parasite replication in both cells and villous explants treated with silver nanoparticles and classical treatment. The AgNp-Bio treatment increased of IL-4 and IL-10 by BeWo cells, while HTR8/SVneo cells produced macrophage migration inhibitory factor (MIF) and IL-4. In the presence of T. gondii, the treatment induced high levels of MIF production by BeWo cells and IL-6 by HTR8SV/neo. In villous explants, the AgNp-Bio treatment downregulated production of IL-4, IL-6, and IL-8 after infection. In conclusion, AgNp-Bio can decrease T. gondii infection in trophoblast cells and villous explants. Therefore, this treatment demonstrated the ability to reduce the T. gondii proliferation with induction of inflammatory mediators in the cells and independent of mediators in chorionic villus which we consider the use of AgNp-Bio promising in the treatment of toxoplasmosis in BeWo and HTR8/SVneo cell models and in chorionic villi.
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- 2020
15. IgE AND IgG antibody responses to Dermatophagoides pteronyssinus in dogs with demodicosis and atopic dermatitis
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José Roberto Mineo, Fabiana Parreira Souza, José Eugênio Diniz Bastos, Ernesto Akio Taketomi, Maria Cecília Oliveira, Neide M. Silva, Deise A. O. Silva, Ana C. A. M. Pajuaba, Marcos Paulo Oliveira Almeida, and Ester Cristina Borges Araujo
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dogs ,Cães ,QH301-705.5 ,demodicosis ,Demodicose ,Immunoglobulin E ,Demodex canis ,Dogs ,IgG. IgE ,Demodicosis ,Medicine ,Biology (General) ,biology ,business.industry ,Agriculture ,Atopic dermatitis ,Dermatophagoides pteronyssinus ,medicine.disease ,Veterinary ,Antibody response ,dermatophagoides pteronyssinus ,igg. ige ,Immunology ,biology.protein ,demodex canis ,General Agricultural and Biological Sciences ,business - Abstract
Canine demodicosis is a common inflammatory parasitic skin disease caused by Demodex mites. House dust mites, such as Dermatophagoides spp., play an important role in the pathogenesis of canine atopic dermatitis (AD). The goal of this experimental work was to investigate whether demodectic dogs could be previously exposed/sensitized to house dust mites’ antigens. First the prevalence of demodicosis in a southeastern region of Brazil was investigated by analyzing clinical files of dogs that were admitted to a Veterinary Hospital. Subsequently, the IgG responses to Dermatophagoides pteronyssinus (Dp) and Dermatophagoides farinae (Df) and IgE to D. pteronyssinus (Dp) were evaluated in two groups, AD or demodicosis dogs. Additionally, the major IgE-binding Dp proteins that are recognized by sera from dogs with demodicosis and AD were evaluated. A total of 2,599 clinical files were analyzed to identify the major parasitic skin diseases in dogs from this region, considering the age, sex and breed of the animals. The epidemiological study identified 111 animals with skin diseases; from these 20.7% presented demodicosis. Afterwards, serum samples were obtained from another groups of demodicosis, AD, and healthy dogs, and analyzed for Dp and Df-specific IgG, and IgE antibody levels, Dp IgG avidity by ELISA and IgE-binding Dp-specific proteins by immunoblot. IgG and IgE antibodies to Dp were detected in sera from additional groups of dogs with AD, demodicosis or healthy, with higher IgE levels to Dp in AD than demodectic or healthy dogs. IgG to Df was detected, despite with smaller levels compared to Dp in sera from demodectic dogs, and also in healthy dogs. Immunoblot showed IgE-binding to Dp proteins in sera of dogs with demodicosis and AD; with strong reactivity for the 72 and 116 kDa antigens detected by sera from demodicosis dogs. However, sera from healthy dogs >12 months old also presented reactivity to these bands. In conclusion, the detection of Dp-IgG and IgE antibodies in sera from demodectic dogs indicates previous exposure and sensitization to the house dust mite, respectively, more than cross-reactivity between demodex mites and Dp antigens detected by canine antibodies. Additionally, higher Dp-specific IgE levels were found in dogs with AD compared with those with demodicosis or healthy, suggesting that Dp-specific IgE could better discriminate dogs with AD from healthy ones or even those with demodicosis. Demodicose canina é uma doença inflamatória comum da pele causada por ácaros do gênero Demodex. Ácaros da poeira doméstica como Dermatophagoides spp. desempenham papel importante na patogênese da dermatite atópica canina (DA). O objetivo desse trabalho experimental foi investigar se cães com demodicose poderiam ser previamente expostos/sensibilizados com antígenos de ácaros da poeira doméstica. A princípio, investigou-se a prevalência de demodicose em uma região sudeste do Brasil, analisando-se prontuários clínicos de cães admitidos em um Hospital Veterinário. Posteriormente, as respostas de IgG a Dermatophagoides pteronyssinus (Dp) e D. farinae (Df) e IgE a D. pteronyssinus (Dp) foram avaliadas em dois grupos, DA ou demodicose. Também foram avaliadas as principais proteínas Dp reconhecidas por anticorpo IgE presente em soros de cães com demodicose e DA. Um total de 2.599 prontuários clínicos foram analisados para identificar as principais doenças parasitárias da pele em cães dessa região, considerando a idade, sexo e raça dos animais. O estudo epidemiológico detectou 111 animais com doenças de pele e destes, 20,7% apresentavam demodicose. Posteriormente, amostras de soro foram obtidas de outros grupos de cães com demodicose, DA ou saudáveis, e analisadas quanto aos níveis de IgG e IgE específicos para Dp e Df, avidez de IgG a Dp por ELISA e proteínas específicas de Dp reconhecidas por IgE por immunoblot. Anticorpos IgG e IgE para Dp foram detectados em soros de grupos adicionais de cães com DA, demodicose ou saudáveis, com níveis mais altos de IgE para Dp na DA do que no soro de animais saudáveis. Níveis de IgG específicos para Df foram detectados, apesar serem menores em comparação com os detectados para Dp em soros de cães demodéticos, e também em cães saudáveis. A análise de immunoblot demonstrou detecção de IgE para proteinas de Dp em soros de cães com demodicose e DA; com forte reatividade para os antígenos de 72 e 116 kDa detectados por soros de cães com demodicose. No entanto, soros de cães saudáveis > 12 meses de idade também apresentaram reatividade a essas bandas. Em conclusão, a detecção de anticorpos Dp-IgG e IgE específicos em soros de cães demodéticos indica exposição prévia e sensibilização aos ácaros, respectivamente, mais do que reatividade cruzada entre ácaros Demodex e antígenos Dp detectados por anticorpos caninos. Além disso, níveis de Dp-IgE específicos mais elevados encontrados em cães com DA, sugerem que esses anticorpos poderiam discriminar melhor cães com DA daqueles saudáveis ou mesmo demodéticos.
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- 2020
16. Establishing tools for early diagnosis of congenital toxoplasmosis: Flow cytometric IgG avidity assay as a confirmatory test for neonatal screening
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Eloisa Amália Vieira Ferro, Elenice Moreira Lemos, Samantha Ribeiro Béla, Ana Carolina Aguiar Vasconcelos Carneiro, Ricardo Wagner de Almeida Vitor, Andréa Teixeira-Carvalho, José Nélio Januário, Anderson Silva Machado, Laura Néspoli Nassar Pansini de Jesus, Aline de Castro Zacche-Tonini, Geisa Baptista Barros, Jordana Grazziela Coelho-dos-Reis, Olindo Assis Martins-Filho, Giuliana Schmidt França Fonseca, José Roberto Mineo, Daniel Vitor Vasconcelos-Santos, and Gláucia Manzan Queiroz Andrade
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0301 basic medicine ,Time Factors ,030231 tropical medicine ,030106 microbiology ,Immunology ,Antibody Affinity ,Antibodies, Protozoan ,Toxoplasmosis, Congenital ,Immunoglobulin G ,Serology ,Flow cytometry ,03 medical and health sciences ,Neonatal Screening ,0302 clinical medicine ,Predictive Value of Tests ,parasitic diseases ,medicine ,Humans ,Immunology and Allergy ,Serologic Tests ,Prospective Studies ,Prospective cohort study ,Whole blood ,biology ,medicine.diagnostic_test ,Infant, Newborn ,Case-control study ,Infant ,Reproducibility of Results ,Flow Cytometry ,Early Diagnosis ,Immunoglobulin M ,Case-Control Studies ,Predictive value of tests ,Host-Pathogen Interactions ,biology.protein ,Dried Blood Spot Testing ,Toxoplasma ,Biomarkers - Abstract
The aim of this study was to evaluate the performance of conventional serology (Q-Preven™ and ELFAVIDAS™) and flow cytometry-based serologic tools for early serologic diagnosis of congenital toxoplasmosis. The study groups included prospectively confirmed cases of congenital toxoplasmosis (TOXO=88) and age-matching non-infected controls (NI=15).The results demonstrated that all samples tested positive/indeterminate for anti-T. gondii IgM screening at birth using air-dried whole blood samples. Serum samples collected at 30-45days after birth tested positive for ELFAVIDAS™ IgG in both groups. While all NI tested negative for ELFAVIDAS™ IgM and IgA, only 78% and 36% of TOXO tested positive for IgM and IgA, respectively. Flow cytometry-based anti-T. gondii IgM, IgA and IgG reactivity displayed moderate performance with low sensitivity (47.6%, 72.6% and 75.0%, respectively). Regardless the remarkable specificity of IgG1, IgG2 and IgG3 subclasses for early diagnosis, weak or moderate specificity was observed (Se=73.9%, 60.2% and 83.0%, respectively). The analysis of IgG avidity indices (AI) demonstrated the highest performance among the flow cytometry-based methods (Se=96.6%; Sp=93.3%), underscoring the low avidity index (AI60%) within TOXO (97.0%) in contrast with the high avidity index (AI60%) in NI (93%). Analysis of anti-T. gondii IgG and IgG3 reactivity for mother:infant paired samples may represent a relevant complementary tests for early diagnosis. In conclusion, a feasible high-standard algorithm (Accuracy=97.1%) was proposed consisting of Q-Preven™ IgM screening at birth, followed by ELFAVIDAS™ IgM and flow cytometric IgG avidity analysis at 30-45days after birth as a high performance tool for early serological diagnosis of congenital toxoplasmosis.
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- 2017
17. Behavioral alterations in long-term Toxoplasma gondii infection of C57BL/6 mice are associated with neuroinflammation and disruption of the blood brain barrier
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Helton C. Santiago, Leda Castaño Barrios, Joseli Lannes-Vieira, Andrea Alice da Silva, Patrícia M.R. e Silva, Ana Paula Da Silva Pinheiro, Ester Roffê, Ricardo T. Gazzinelli, Daniel Gibaldi, Neide M. Silva, and José Roberto Mineo
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Central Nervous System ,Time Factors ,Physiology ,Social Sciences ,Brain Edema ,Anxiety ,Nervous System ,Toxoplasma Gondii ,Mice ,Medical Conditions ,Immune Physiology ,Medicine and Health Sciences ,Psychology ,Immune Response ,Mammals ,Protozoans ,Innate Immune System ,Multidisciplinary ,Animal Behavior ,Behavior, Animal ,biology ,Depression ,Eukaryota ,Animal Models ,Up-Regulation ,medicine.anatomical_structure ,Experimental Organism Systems ,Blood-Brain Barrier ,Toxoplasmosis, Cerebral ,Vertebrates ,Cytokines ,Medicine ,Female ,Anatomy ,medicine.symptom ,Toxoplasma ,Locomotion ,Research Article ,Science ,Immunology ,Central nervous system ,Mouse Models ,Inflammation ,CCL2 ,Research and Analysis Methods ,Blood–brain barrier ,Rodents ,Model Organisms ,Signs and Symptoms ,parasitic diseases ,Parasitic Diseases ,medicine ,Animals ,Parasites ,Muscle Strength ,Neuroinflammation ,Behavior ,business.industry ,Organisms ,Biology and Life Sciences ,Toxoplasma gondii ,Molecular Development ,medicine.disease ,biology.organism_classification ,Parasitic Protozoans ,Toxoplasmosis ,Mice, Inbred C57BL ,Chronic infection ,Immune System ,Amniotes ,Chronic Disease ,Animal Studies ,Clinical Medicine ,business ,Zoology ,Developmental Biology - Abstract
The Apicomplexa protozoanToxoplasma gondiiis a mandatory intracellular parasite and the causative agent of toxoplasmosis. This illness is of medical importance due to its high prevalence worldwide and may cause neurological alterations in immunocompromised persons. In chronically infected immunocompetent individuals, this parasite forms tissue cysts mainly in the brain. In addition,T.gondiiinfection has been related to mental illnesses such as schizophrenia, bipolar disorder, depression, obsessive-compulsive disorder, as well as mood, personality, and other behavioral changes. In the present study, we evaluated the kinetics of behavioral alterations in a model of chronic infection, assessing anxiety, depression and exploratory behavior, and their relationship with neuroinflammation and parasite cysts in brain tissue areas, blood-brain-barrier (BBB) integrity, and cytokine status in the brain and serum. Adult female C57BL/6 mice were infected by gavage with 5 cysts of the ME-49 type IIT.gondiistrain, and analyzed as independent groups at 30, 60 and 90 days postinfection (dpi). Anxiety, depressive-like behavior, and hyperactivity were detected in the early (30 dpi) and long-term (60 and 90 dpi) chronicT.gondiiinfection, in a direct association with the presence of parasite cysts and neuroinflammation, independently of the brain tissue areas, and linked to BBB disruption. These behavioral alterations paralleled the upregulation of expression of tumor necrosis factor (TNF) and CC-chemokines (CCL2/MCP-1, CCL3/MIP-1α, CCL4/MIP-1β and CCL5/RANTES) in the brain tissue. In addition, increased levels of interferon-gamma (IFNγ), TNF and CCL2/MCP-1 were detected in the peripheral blood, at 30 and 60 dpi. Our data suggest that the persistence of parasite cysts induces sustained neuroinflammation, and BBB disruption, thus allowing leakage of cytokines of circulating plasma into the brain tissue. Therefore, all these factors may contribute to behavioral changes (anxiety, depressive-like behavior, and hyperactivity) in chronicT.gondiiinfection.
- Published
- 2021
18. ERK1/2 phosphorylation and IL-6 production are involved in the differential susceptibility to Toxoplasma gondii infection in three types of human (cyto/ syncytio/ extravillous) trophoblast cells
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Samuel Cota Teixeira, Mayara Ribeiro, Eloisa Amália Vieira Ferro, Rafaela José da Silva, A.O. Gomes, Priscila Silva Franco, B.F. Barbosa, Pamela Mendonça Guirelli, A.S. Castro, F.C. Oliveira, and José Roberto Mineo
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0301 basic medicine ,Cell type ,Population ,Giant Cells ,Andrology ,03 medical and health sciences ,0302 clinical medicine ,Syncytiotrophoblast ,Cell Line, Tumor ,parasitic diseases ,medicine ,Humans ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,education ,Interleukin 6 ,reproductive and urinary physiology ,Cell Proliferation ,education.field_of_study ,Fetus ,030219 obstetrics & reproductive medicine ,Cytotrophoblast ,biology ,Interleukin-6 ,Trophoblast ,Toxoplasma gondii ,Cell Differentiation ,Cell Biology ,General Medicine ,biology.organism_classification ,Trophoblasts ,Up-Regulation ,030104 developmental biology ,medicine.anatomical_structure ,embryonic structures ,biology.protein ,Disease Susceptibility ,Toxoplasmosis ,Developmental Biology - Abstract
During pregnancy, Toxoplasma gondii can triggers serious manifestations and potentially affect the fetal development. In this scenario, differences in susceptibility of trophoblast cells to T. gondii infection might be evaluated in order to establish new therapeutic approaches capable of interfering in the control of fetal infection by T. gondii. This study aimed to evaluate the susceptibility of cytotrophoblast, syncytiotrophoblast and extravillous trophoblast cells to T. gondii infection. Our data demonstrate that HTR-8/SVneo cells (extravillous trophoblast cells) present higher susceptibility to T. gondii infection when compared to syncytiotrophoblast and cytotrophoblast cells, whereas syncytiotrophoblast was the cell type more resistant to the parasite infection. Also, cytotrophoblast and syncytiotrophoblast cells produced significantly more IL-6 than HTR-8/SVneo cells. On the other hand, HTR-8/SVneo cells showed higher ERK1/2 phosphorylation than cytotrophoblast and syncytiotrophoblast cells. ERK1/2 inhibition reduced T. gondii infection and increased IL-6 production in HTR-8/SVneo cells. Thus, it is plausible to conclude that the greater susceptibility of HTR-8/SVneo cells to infection by T. gondii is related to a higher ERK1/2 phosphorylation and lower levels of IL-6 in these cells compared to other cells, suggesting that these mediators may be important to favor the parasite infection in this type of trophoblastic population.
- Published
- 2021
19. A peptide originated from Toxoplasma gondii microneme 8 displaying serological evidence to differentiate recent from chronic human infection
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Ana C. A. M. Pajuaba, Fernando R. Carvalho, Vinícius Fernandes Paiva, Reynaldo Dietze, Tamires Lopes Silva, José Roberto Mineo, Patrício da Silva Cardoso Barros, Heber Leão Silva Barros, Geisa Baptista Barros, Silas Silva Santana, and Tiago W. P. Mineo
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Protozoan Proteins ,Serological evidence ,Peptide ,Disease ,Microneme ,Mice ,Antigen ,Seroepidemiologic Studies ,medicine ,Animals ,Humans ,Serologic Tests ,chemistry.chemical_classification ,Mice, Inbred BALB C ,biology ,business.industry ,Toxoplasma gondii ,biology.organism_classification ,medicine.disease ,Toxoplasmosis ,Infectious Diseases ,chemistry ,Immunology ,biology.protein ,Female ,Parasitology ,Antibody ,Peptides ,business ,Cell Adhesion Molecules ,Toxoplasma ,Biomarkers - Abstract
Toxoplasmosis is able to cause death and/or sequelae in foetuses from pregnant women and immunocompromised individuals. The early diagnosis, able to differentiate acute from chronic phases, is essential to define the treatment against this disease and minimize the risk of complications. Here we describe a peptide derived from microneme 8 (pMIC8) protein of Toxoplasma gondii, able to distinguish the phase of infection. By using human and mice serum samples with different infection times, we assessed the ability of pMIC8 to interact with antibodies present in early of infection, and compared the results obtained with soluble antigen of T. gondii (STAg). The results showed that pMIC8 was recognized more precisely with antibodies present in serum samples from individuals with time of infection below 3 months, followed by those between 4 and 6 months of infection. Based on these results, it is possible to conclude that the association of immunoassays using STAg and pMIC8 as antigen preparations can be used to distinguish acute from chronic infections.
- Published
- 2021
20. Randomized Controlled Trial of Oropharyngeal Colostrum Administration in Very-low-birth-weight Preterm Infants
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Morun Bernardino Neto, Vânia Olivetti Steffen Abdallah, Fernanda Maria Santiago, Giovana G Fatureto, Nathália G Afonso, Natássia F Navarro, Daniela Marques de Lima Mota Ferreira, Débora V de Leves, Francisco Eulógio Martinez, Mônica Camargo Sopelete, Angela Maria de Morais Oliveira, José Roberto Mineo, and Érica B de Bem
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Immunoglobulin A ,Male ,medicine.medical_specialty ,Gestational Age ,Breast milk ,Placebo ,Gastroenterology ,law.invention ,Sepsis ,03 medical and health sciences ,0302 clinical medicine ,Enteral Nutrition ,Randomized controlled trial ,Double-Blind Method ,law ,030225 pediatrics ,Internal medicine ,Medicine ,Humans ,Infant, Very Low Birth Weight ,biology ,Milk, Human ,business.industry ,Colostrum ,Infant, Newborn ,Gestational age ,medicine.disease ,Low birth weight ,Breast Feeding ,Immunoglobulin G ,IMUNOTERAPIA ,Pediatrics, Perinatology and Child Health ,biology.protein ,030211 gastroenterology & hepatology ,Female ,medicine.symptom ,Neonatal Sepsis ,business - Abstract
Objective The purpose of this study was to evaluate the effects of oropharyngeal colostrum administration in the incidence of late-onset clinical and proven sepsis and in concentrations of immunoglobulin A (IgA) in very-low-birth-weight (VLBW) infants. Methods We conducted a double-blinded, randomized, placebo-controlled trial and assigned 113 VLBW infants to receive 0.2 mL of maternal colostrum or sterile water (placebo) via oropharyngeal route every 2 hours for 48 hours, beginning in the first 48 to 72 hours of life. Neonates of both groups were fed breast milk from the first 3 days of life until a volume of at least 100 mL · kg · day. IgA was measured in serum and urine before and after treatment. Clinical data during hospitalization were collected. Results We found no statistically significant differences between colostrum and placebo groups in the incidence of late-onset clinical sepsis (odds ratio 0.7602; CI 95% 0.3-1.6) and proven sepsis (odds ratio 0.7028; CI 95% 0.3-1.6). The measurement of IgA was similar in serum before (P value 0.87) and after treatment (P value 0.26 day 4 and 0.77 day 18). No differences were also observed in IgA in urine before (P value 0.8) and after treatment (P value 0.73 day 4 and 0.52). Conclusions This study could not confirm the hypothesis that oropharyngeal administration of maternal colostrum to VLBW could reduce the incidence of late-onset sepsis and increase the levels of IgA. We believe that this finding can be justified by the practice of feeding VLBW infants exclusively with breast milk in the first days of life and reinforces the prior knowledge of the importance of early nutrition, especially, with human milk. It also suggests that oropharyngeal administration of colostrum should be reserved for neonates who cannot be fed in first few days of life.
- Published
- 2019
21. Toll-Like Receptor 3–TRIF Pathway Activation by Neospora caninum RNA Enhances Infection Control in Mice
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Vanessa dos Santos Miranda, Fernanda Maria Santiago, Vanessa Resende Souza Silva, Patrício da Silva Cardoso Barros, Caroline M. Mota, Flávia Batista Ferreira França, Mylla Spirandelli da Costa, Kleber Simônio Parreira, Tiago W. P. Mineo, and José Roberto Mineo
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0301 basic medicine ,Toll-like receptor ,biology ,medicine.medical_treatment ,Immunology ,Toxoplasma gondii ,biology.organism_classification ,Microbiology ,Neospora caninum ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Infectious Diseases ,Cytokine ,TRIF ,Interferon ,parasitic diseases ,medicine ,Parasitology ,Tumor necrosis factor alpha ,IRF3 ,030215 immunology ,medicine.drug - Abstract
Neospora caninum is a protozoan parasite closely related to Toxoplasma gondii and has been studied for causing neuromuscular disease in dogs and abortions in cattle. It is recognized as one of the main transmissible causes of reproductive failure in cattle and consequent economic losses to the sector. In that sense, this study aimed to evaluate the role of Toll-like receptor 3 (TLR3)-TRIF-dependent resistance against N. caninum infection in mice. We observed that TLR3-/- and TRIF-/- mice presented higher parasite burdens, increased inflammatory lesions, and reduced production of interleukin 12p40 (IL-12p40), tumor necrosis factor (TNF), gamma interferon (IFN-γ), and nitric oxide (NO). Unlike those of T. gondii, N. caninum tachyzoites and RNA recruited TLR3 to the parasitophorous vacuole (PV) and translocated interferon response factor 3 (IRF3) to the nucleus. We also observed that N. caninum upregulated the expression of TRIF in murine macrophages, which in turn upregulated IFN-α and IFN-β in the presence of the parasite. Furthermore, TRIF-/- infected macrophages produced lower levels of IL-12p40, while exogenous IFN-α replacement was able to completely restore the production of this key cytokine. Our results show that the TLR3-TRIF signaling pathway enhances resistance against N. caninum infection in mice, since it improves Th1 immune responses that result in controlled parasitism and reduced tissue inflammation, which are hallmarks of the disease.
- Published
- 2019
22. Acetonic Fraction of Bidens pilosa Enriched for Maturase K Is Able to Control Cerebral Parasite Burden in Mice Experimentally Infected With Toxoplasma gondii
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Taísa Carrijo de Oliveira, Carlos Priminho Pirovani, Deise A. O. Silva, Mariana R.D. Cardoso, Caroline M. Mota, Cristina Rostkowska, José Roberto Mineo, Fernanda Maria Santiago, and Tiago W. P. Mineo
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040301 veterinary sciences ,ved/biology.organism_classification_rank.species ,Toxoplasma gondii ,Biology ,Microbiology ,0403 veterinary science ,03 medical and health sciences ,In vivo ,Bidens pilosa ,parasitic diseases ,medicine ,Parasite hosting ,Maturase K ,Original Research ,030304 developmental biology ,0303 health sciences ,lcsh:Veterinary medicine ,General Veterinary ,ved/biology ,04 agricultural and veterinary sciences ,biology.organism_classification ,medicine.disease ,infection control ,In vitro ,Toxoplasmosis ,maturase k ,Toxicity ,lcsh:SF600-1100 ,Veterinary Science ,total and acetonic extracts - Abstract
Toxoplasma gondii infection can cause abortions or congenital infection for a vast number of domestic animals and humans, leading to economic loss in veterinary sciences, as well as severe consequences for immunocompromised patients. Bidens pilosa Linné has been used in ethnopharmacology for treatment of diseases, as malaria, diabetes and hepatitis, in addition to its use as antioxidant, antiallergic, anti-inflammatory, and antiviral. The components of this plant have never been studied before for treatment of toxoplasmosis, and the conventional drugs currently used to treat this disease have high degree of toxicity. Thus, the aim of this study was to evaluate the effect of B. pilosa against T. gondii, by analyzing a total extract of this plant in parallel with a fraction obtained by precipitation in acetone. Also, it was assessed if the acetonic fraction could present lectinic activity, followed by its identification by mass spectrometry. It was observed with the experimental models designed that both total extract and acetonic fraction of B. pilosa were able to control T. gondii infection by in vitro and in vivo experiments, in addition to their low toxicity to host cells. Both total extract and acetonic fraction of this plant display capacity to impair replication of T. gondii tachyzoites. Interesting, the B. pilosa acetonic fraction treatment for 10 days after infection decreases significantly the number of T. gondii brain cyst in comparison with controls. The protein isolated from B. pilosa acetonic fraction was characterized as a novel lectin identified as maturase K. Taken together, these findings open new perspectives to treat patients infected by T. gondii. Future studies will be necessary to investigate the precise mechanism underlying the control of T. gondii infection to impair the replication of this parasite in the host cells after treatment with B. pilosa maturase K.
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- 2019
23. C57BL/6 mice immunized with synthetic peptides from Toxoplasma gondii surface and microneme immunodominant antigens are able to decrease parasite burden in the brain tissues
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Heber Leão Silva Barros, Patrício da Silva Cardoso Barros, Silas Silva Santana, Fernando R. Carvalho, Ana C. A. M. Pajuaba, José Roberto Mineo, Tiago W. P. Mineo, and Vinícius Fernandes de Paiva
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0301 basic medicine ,Protozoan Vaccines ,Veterinary (miscellaneous) ,030231 tropical medicine ,Protozoan Proteins ,Antibodies, Protozoan ,Antigens, Protozoan ,Biology ,Microbiology ,Microneme ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Immune system ,Antigen ,parasitic diseases ,medicine ,Parasite hosting ,Animals ,Rhoptry ,Immunodominant Epitopes ,Toxoplasma gondii ,Brain ,030108 mycology & parasitology ,medicine.disease ,biology.organism_classification ,Toxoplasmosis ,Mice, Inbred C57BL ,Infectious Diseases ,Toxoplasmosis, Animal ,Insect Science ,biology.protein ,Cytokines ,Epitopes, B-Lymphocyte ,Parasitology ,Female ,Immunization ,Antibody ,Peptides ,Toxoplasma - Abstract
Toxoplasmosis is a disease caused by Toxoplasma gondii, an intracellular protozoan able to infect a wide range of hosts. The infection is particularly severe in immunocompromised patients or during pregnancy, circumstances in which the parasite could find a more favorable microenvironment to replicate and invade host tissues. The current treatment consists in toxic drugs for the patients, being not appropriate for the fetuses and immunodeficient patients. So far, there is a lack of available vaccine to prevent the disease. The present study aimed to evaluate the immune response induced by peptides derived from parasite immunodominant proteins from key components, as surface, rhoptry, microneme and dense granule antigens. A panel of eleven peptides was selected considering the highest scores for B cell epitope prediction by in silico analyses. The peptides were divided in groups, according to the parasite organelle locations, and used to immunize C57BL/6 mice. The animals were submitted to three doses of immunization and infected by 10 cysts of T. gondii ME49 strain. Blood samples were collected and used to measure the production of antibodies and cytokines, while the brains were collected to determine the parasite burden by quantitative polymerase chain reaction (qPCR). It was found that synthetic peptides from all targets were able to induce IgG synthesis in immunized mice, as well as to modulate the Th1/Th2 cytokine production, particularly the MIC and SRS groups, which presented the IFN-γ/IL-10 and TNF-α/IL-10 ratios 30 and 10 times higher, respectively, when compared with non-immunized group. Interestingly, the animals from MIC and SRS groups had significantly lower levels of T. gondii DNA in their brains. In summary, it can be concluded that peptides mainly from SRS and MIC parasite components constitute relevant targets to design vaccine candidates against parasite burden observed during chronic toxoplasmosis.
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- 2019
24. Cyclooxygenase (COX)-2 Inhibitors Reduce
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Ana Carolina Alcântara, Pereira, Rafaela José, Silva, Priscila Silva, Franco, Angelica, de Oliveira Gomes, Guilherme, Souza, Iliana Claudia Balga, Milian, Mayara, Ribeiro, Alessandra Monteiro, Rosini, Pâmela Mendonça, Guirelli, Eliézer Lucas Pires, Ramos, Tiago Wilson Patriarca, Mineo, José Roberto, Mineo, Neide Maria, Silva, Eloisa Amália Vieira, Ferro, and Bellisa Freitas, Barbosa
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prostaglandin E2 ,Calomys callosus ,cyclooxygenase-2 ,parasitic diseases ,THP-1 cells ,Toxoplasma gondii ,Microbiology ,immune response ,Original Research - Abstract
Toxoplasma gondii is able to infect a wide range of vertebrates, including humans. Studies show that cyclooxygenase-2 (COX-2) is a modulator of immune response in multiple types of infection, such as Trypanosoma cruzi. However, the role of COX-2 during T. gondii infection is still unclear. The aim of this study was to investigate the role of COX-2 during infection by moderately or highly virulent strains of T. gondii in Calomys callosus rodents and human THP-1 cells. C. callosus were infected with 50 cysts of T. gondii (ME49), treated with COX-2 inhibitors (meloxicam or celecoxib) and evaluated to check body weight and morbidity. After 40 days, brain and serum were collected for detection of T. gondii by real-time PCR and immunohistochemistry or cytokines by CBA. Furthermore, peritoneal macrophages or THP-1 cells, infected with RH strain or uninfected, were treated with meloxicam or celecoxib to evaluate the parasite proliferation by colorimetric assay and cytokine production by ELISA. Finally, in order to verify the role of prostaglandin E2 in COX-2 mechanism, THP-1 cells were infected, treated with meloxicam or celecoxib plus PGE2, and analyzed to parasite proliferation and cytokine production. The data showed that body weight and morbidity of the animals changed after infection by T. gondii, under both treatments. Immunohistochemistry and real-time PCR showed a reduction of T. gondii in brains of animals treated with both COX-2 inhibitors. Additionally, it was observed that both COX-2 inhibitors controlled the T. gondii proliferation in peritoneal macrophages and THP-1 cells, and the treatment with PGE2 restored the parasite growth in THP-1 cells blocked to COX-2. In the serum of Calomys, upregulation of pro-inflammatory cytokines was detected, while the supernatants of peritoneal macrophages and THP-1 cells demonstrated significant production of TNF and nitrite, or TNF, nitrite and MIF, respectively, under both COX-2 inhibitors. Finally, PGE2 treatment in THP-1 cells triggered downmodulation of pro-inflammatory mediators and upregulation of IL-8 and IL-10. Thus, COX-2 is an immune mediator involved in the susceptibility to T. gondii regardless of strain or cell types, since inhibition of this enzyme induced control of infection by upregulating important pro-inflammatory mediators against Toxoplasma.
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- 2018
25. Brazilian strains of Toxoplasma gondii are controlled by azithromycin and modulate cytokine production in human placental explants
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José Roberto Mineo, B.F. Barbosa, Thádia Evelyn de Araújo, Rafaela José da Silva, Eloisa Amália Vieira Ferro, Priscila Silva Franco, A.O. Gomes, Paula Suellen Guimarães Gois, Lara Affonso dos Santos, Francesca Ietta, and Maria Célia dos Santos
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0301 basic medicine ,Transplacental transmission ,Endocrinology, Diabetes and Metabolism ,Placenta ,Pregnancy Trimester, Third ,Clinical Biochemistry ,Virulence ,lcsh:Medicine ,Toxoplasma gondii ,Azithromycin ,Microbiology ,03 medical and health sciences ,0302 clinical medicine ,Pregnancy ,Genotype ,parasitic diseases ,medicine ,Parasite hosting ,Humans ,Pharmacology (medical) ,Molecular Biology ,Third ,biology ,Research ,Biochemistry (medical) ,Spiramycin ,lcsh:R ,Cell Biology ,General Medicine ,Brazilian strains ,biology.organism_classification ,Villous explants ,Brazil ,Coccidiostats ,Cytokines ,Female ,Toxoplasma ,030104 developmental biology ,Pyrimethamine ,030220 oncology & carcinogenesis ,Pregnancy Trimester ,medicine.drug - Abstract
Background Toxoplasma gondii is a protozoan parasite that causes congenital toxoplasmosis by transplacental transmission. Parasite strains are genetically diverse and disease severity is related to the genotype. In Uberlândia city, Brazil, two virulent strains were isolated: TgChBrUD1 and TgChBrUD2. Congenital toxoplasmosis is more prevalent in South America compared to Europe, and more often associated with severe symptoms, usually as a result of infection with atypical strains. Methods Considering that T. gondii has shown high genetic diversity in Brazil, the effectiveness of traditional treatment may not be the same, as more virulent strains of atypical genotypes may predominate. Thus, the aim of this study were to evaluate the Brazilian strain infection rate in human villous explants and the azithromycin efficacy with regard to the control of these strains compared to traditional therapy. Villi were infected with RH, ME49, TgChBrUD1 or TgChBrUD2 strains and treated with azithromycin, spiramycin or a combination of pyrimethamine plus sulfadiazine. The villous viability was analyzed by LDH assay and morphological analysis. Parasite proliferation, as well as production of cytokines was analyzed by qPCR and ELISA, respectively. Statistical analysis was performed using the GraphPad Prism 5.0. Results The treatments were not toxic and TgChBrUD1 infected villi showed a higher parasite burden compared with others strains. Treatments significantly reduced the intracellular proliferation of T. gondii, regardless of the strain. TgChBrUD1-infected villi produced a larger amount of MIF, IL-6 and TGF-β1 compared with other infected villi. Azithromycin treatment increased MIF production by RH- or TgChBrUD2-infected villi, but in ME49- or TgChBrUD1-infected villi, the MIF production was not altered by treatment. On the other hand, azithromycin treatment induced lower IL-6 production by ME49- or TgChBrUD1-infected villi. Conclusions Azithromycin treatment was effective against T. gondii Brazilian strains compared with conventional treatment. Also, the TgChBrUD1 strain replicated more in villi and modulated important cytokines involved in parasite control, showing that different strains use different strategies to evade the host immune response and ensure their survival.
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- 2018
26. Macrophage migration inhibitory factor (MIF) and pregnancy may impact the balance of intestinal cytokines and the development of intestinal pathology caused by Toxoplasma gondii infection
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Marcos de Lucca Moreira Gomes, Priscila Silva Franco, Camila Ferreira Marcon, Eloisa Amália Vieira Ferro, Rafaela José da Silva, B.F. Barbosa, Paula Tatiana Mutão Ferreira, José Roberto Mineo, Carlo José Freire Oliveira, Mayara Ribeiro, Tiago W. P. Mineo, Javier Emílio Lazo Chica, Virmondes Rodrigues Junior, Roberto Augusto Pereira de Sousa, and A.O. Gomes
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,Duodenum ,medicine.medical_treatment ,Immunology ,Ileum ,Biology ,Biochemistry ,Pathogenesis ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Pregnancy ,medicine ,Animals ,Immunology and Allergy ,Macrophage Migration-Inhibitory Factors ,Molecular Biology ,Mice, Knockout ,Toxoplasma gondii ,Hematology ,medicine.disease ,biology.organism_classification ,Toxoplasmosis ,Small intestine ,Intramolecular Oxidoreductases ,030104 developmental biology ,medicine.anatomical_structure ,Cytokine ,Pregnancy Complications, Parasitic ,030220 oncology & carcinogenesis ,Female ,Macrophage migration inhibitory factor ,Toxoplasma - Abstract
Toxoplasma gondii (T. gondii) is an intracellular parasite responsible for causing toxoplasmosis. When infection occurs during pregnancy, it can produce severe congenital infection with ocular and neurologic damage to the infant. From the oral infection parasite reaches the intestine, causing inflammatory response, damage in tissue architecture and systemic dissemination. Macrophage migration inhibition factor (MIF) is a cytokine secreted from both immune and non-immune cells, including gut epithelial cells. MIF is described to promote inflammatory responses, to be associated in colitis pathogenesis and also to play role in maintaining the intestinal barrier. The aim of the present study was to evaluate the influence of the pregnancy and MIF deficiency on T. gondii infection in the intestinal microenvironment and to address how these factors can impact on the intestinal architecture and local cytokine profile. For this purpose, small intestine of pregnant and non-pregnant C57BL/6 MIF deficient mice (MIF-/-) and Wild-type (WT) orally infected with 5 cysts of ME-49 strain of T. gondii were collected on day 8th of infection. Intestines were processed for morphological and morphometric analyses, parasite quantification and for cytokines mensuration. Our results showed that the absence of MIF and pregnancy caused an increase in T. gondii infection index. T. gondii immunolocalization demonstrated that segments preferentially infected with T. gondii were duodenum and ileum. The infection caused a reduction in the size of the intestinal villi, whereas, infection associated with pregnancy caused an increase in villi size due to edema caused by the infection. Also, the goblet cell number was increased in the ileum of MIF-/- mice, when compared to the corresponding WT group. Analyses of cytokine production in the small intestine showed that MIF was up regulated in the gut of pregnant WT mice due to infection. Also, infection provoked an intense Th1 response that was more exacerbated in pregnant MIF-/- mice. We also detected that the Th2/Treg response was more pronounced in MIF-/- mice. Altogether, our results demonstrated that pregnancy and MIF deficiency interferes in the balance of the intestinal cytokines and favors a Th1-immflamatory profile, which in turn, impact in the development of pathology caused by T. gondii infection in the intestinal microenvironment.
- Published
- 2020
27. Isolation, genetic and immunohistochemical identification of Toxoplasma gondii from human placenta in a large toxoplasmosis outbreak in southern Brazil, 2018
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Fernanda Silveira Flores Vogel, Selwyn Arlington Headley, Patricia Bräunig, Aline Ludwig, José Roberto Mineo, Roberta Lemos Freire, Regina Mitsuka-Breganó, Ivone Andreatta Menegolla, Cledison Marcio Difante, Natalia Canal, Luís Antônio Sangioni, Lourdes Bonfleur Farinha, Kerlei Cristina Médici, Beatriz de Souza Lima Nino, Italmar Teodorico Navarro, Camila Encarnação Minuzzi, Luiza Pires Portella, Isadora Cortella Britto, Simone Haas, Luciane Silva Ramos, Fernanda Pinto-Ferreira, Felippe Danyel Cardoso Martins, Liliane Souto Pacheco, João Luis Garcia, Camila Ribeiro Silva, Ariana Signori, Flávia Caselli Pacheco, and Thais Cabral Monica
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0301 basic medicine ,Microbiology (medical) ,Amniotic fluid ,Placenta ,030106 microbiology ,Environment ,Microbiology ,Disease Outbreaks ,law.invention ,03 medical and health sciences ,Pregnancy ,law ,Genotype ,Genetics ,medicine ,Humans ,Bioassay ,Public Health Surveillance ,Molecular Biology ,Ecology, Evolution, Behavior and Systematics ,Polymerase chain reaction ,biology ,Toxoplasma gondii ,Outbreak ,biology.organism_classification ,medicine.disease ,Virology ,Toxoplasmosis ,Pregnancy Complications ,030104 developmental biology ,Infectious Diseases ,medicine.anatomical_structure ,Female ,Disease Susceptibility ,Toxoplasma ,Brazil - Abstract
The present study aimed to describe a molecular analysis of environmental and pork samples, the isolation, genetic identification and immunohistochemistry (IHC) of Toxoplama gondii from placenta and amniotic fluid from five pregnant women that miscarried during a toxoplasmosis outbreak in 2018, Santa Maria, Rio Grande do Sul. Environmental and pork samples were submitted to polymerase chain reaction (PCR); placenta and amniotic fluid samples to histopathology, IHC, mouse bioassay and PCR. All samples were genotyped by PCR-RFLP with 11 loci. Histopathologic and IHC were compatibles with toxoplasmosis. All pregnants were positive in PCR and bioassay, the genotypes were compared, and all were equal suggesting a same source of infection. Among the environmental and food samples, a sludge sample from a water tank and two porks samples were positive in PCR, and the genotypes were different from the pregnant women isolates. It is concluded that obtain and compare isolates is essential to elucidate outbreak source.
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- 2020
28. Anti-parasitic effect on Toxoplasma gondii induced by BnSP-7, a Lys49-phospholipase A2 homologue from Bothrops pauloensis venom
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Isabela Pacheco Borges, Eloisa Amália Vieira Ferro, Veridiana M. Rodrigues, José Roberto Mineo, Kelly Aparecida Yoneyama Tudini, B.F. Barbosa, Renata Santos Rodrigues, Rafaela José da Silva, Dayane Lorena Naves de Souza, and Letícia Eulalio Castanheira
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0301 basic medicine ,030106 microbiology ,Toxicology ,medicine.disease_cause ,Microbiology ,HeLa ,03 medical and health sciences ,chemistry.chemical_compound ,Immune system ,Crotalid Venoms ,medicine ,Humans ,MTT assay ,Amino Acid Sequence ,Cytotoxicity ,Chromatography, High Pressure Liquid ,Phospholipase A ,Sequence Homology, Amino Acid ,biology ,Toxin ,Lysine ,Toxoplasma gondii ,biology.organism_classification ,Phospholipases A2 ,030104 developmental biology ,chemistry ,Electrophoresis, Polyacrylamide Gel ,Trypan blue ,Toxoplasma ,HeLa Cells - Abstract
Toxoplasmosis affects a third of the global population and presents high incidence in tropical areas. Its great relevance in public health has led to a search for new therapeutic approaches. Herein, we report the antiparasitic effects of BnSP-7 toxin, a Lys49 phospholipase A2 (PLA2) homologue from Bothrops pauloensis snake venom, on Toxoplasma gondii. In an MTT assay, BnSP-7 presented significant cytotoxicity against host HeLa cells at higher doses (200 μg/mL to 50 μg/mL), whereas lower doses (25 μg/mL to 1.56 μg/mL) produced low cytotoxicity. Furthermore, the toxin showed no effect on T. gondii tachyzoite viability when evaluated by trypan blue exclusion, but decreased both adhesion and parasite proliferation when tachyzoites were treated before infection. We also measured cytokines in supernatants collected from HeLa cells infected with T. gondii tachyzoites previously treated with RPMI or BnSP-7, which revealed enhancement of only MIF and IL-6 cytokines levels in supernatants of HeLa cells after BnSP-7 treatment. Our results showed that the BnSP-7 PLA2 exerts an anti-Toxoplasma effect at a lower dose than that required to induce cytotoxicity in HeLa cells, and also modulates the immune response of host cells. In this sense, the anti-parasitic effect of BnSP-7 PLA2 demonstrated in the present study opens perspectives for use of this toxin as a tool for future studies on toxoplasmosis.
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- 2016
29. IgA and IgG1 reactivities assessed by flow cytometry mirror clinical aspects of infants with ocular congenital toxoplasmosis
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Ana Carolina Aguiar Vasconcelos Carneiro, Samantha Ribeiro Béla, Andréa Teixeira-Carvalho, Eloisa Amália Vieira Ferro, José Nélio Januário, Ricardo Wagner de Almeida Vitor, Anderson Silva Machado, Geisa Baptista Barros, Jordana Grazziela Coelho-dos-Reis, Laura Néspoli Nassar Pansini de Jesus, Lis Ribeiro do Valle Antonelli, Olindo Assis Martins-Filho, Elenice Moreira Lemos, Lilian Maria Garcia Bahia-Oliveira, Aline de Castro Zacche Tonini, Gláucia Manzan Queiroz Andrade, José Roberto Mineo, and Daniel Vitor Vasconcelos-Santos
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0301 basic medicine ,Immunoglobulin A ,Pathology ,medicine.medical_specialty ,medicine.medical_treatment ,030231 tropical medicine ,Immunology ,Toxoplasmosis, Congenital ,Immunoglobulin G ,Serology ,Flow cytometry ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Humans ,Immunology and Allergy ,Prospective Studies ,biology ,medicine.diagnostic_test ,fungi ,Infant ,Flow Cytometry ,Acquired immune system ,Cross-Sectional Studies ,030104 developmental biology ,Cytokine ,biology.protein ,Cytokines ,Antibody ,CD8 - Abstract
This study intended to apply the flow cytometric analysis of IgA and IgG reactivity and intracytoplasmic cytokine analysis to understand and decode the clinical aspects of infants with ocular congenital toxoplasmosis. The Toxoplasma gondii-infected infants (TOXO) were subdivided according to their clinical aspects based on the absence (NRL), presence of active (ARL), active/cicatricial (ACRL) or cicatricial retinochoroidal lesions (CRL) and compared to non-infected controls (NI). The reactivity of anti-T. gondii IgG subclasses resembles the clinical aspects of ocular lesions. IgG and IgG1 discriminate infants with cicatricial lesions (ACRL and CRL) from both ARL and NLR. IgG2 and IgG3 are particularly higher in ACRL and CRL as compared to NLR. No differences were observed when IgG4 reactivity was evaluated. Thus, the results indicated that the reactivity patterns of IgA, IgG and IgG subclasses are able to discriminate ARL, ACRL and CRL from NLR or NI. IgA and IgG subclasses are relevant serological biomarkers with diagnostic and prognostic applicability, respectively. Moreover, IgA and IgG1 were closely related to cytokine production by innate/adaptive immunity cells. IgA reactivity was directly associated to TNF-α-derived from neutrophils, monocytes and CD8(+) T-cells, while IgG1 was inversely correlated with IFN-γ-producing CD4(+) and CD8(+) T-cells but positively correlated with IL-10(+) B-cells. These findings provide insights on the relationship between the cytokine production by innate/adaptive immunity and the antibody pattern of infants with ocular congenital toxoplasmosis. In addition, the present study supports the use of flow cytometric serology as a potential tool for the diagnosis and monitoring of ocular lesions in T. gondii-infected infants in the clinical setting.
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- 2016
30. Treatment with a Zinc Metalloprotease Purified from
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Maraisa Cristina, Silva, Helioswilton, Sales-Campos, Carlo José Freire, Oliveira, Tamires Lopes, Silva, Flávia Batista Ferreira, França, Fábio, Oliveira, Tiago Wilson Patriarca, Mineo, and José Roberto, Mineo
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Mice, Inbred C57BL ,Mice ,Dextran Sulfate ,Animals ,Cytokines ,Metalloendopeptidases ,Bothrops ,Colitis ,Research Article - Abstract
It has been described that the metalloprotease BmooMP-alpha-I purified from Bothrops moojeni snake venom is able to hydrolyze the TNF molecule. However, this observation has been based mainly on in vitro investigation, in addition to molecular modeling and docking approaches. Considering that there is no in vivo study to demonstrate the biological effects of this enzyme, the major aim to the present work was to investigate whether the BmooMP-alpha-I has any anti-inflammatory efficacy by setting up a murine experimental design of colitis induced by dextran sulfate sodium (DSS). For this purpose, C57BL/6 mice were divided into six groups, as follows: (i) animals without intestinal inflammation, (ii) animals without intestinal inflammation treated with BmooMP-alpha-I (50 μg/animal/day), and (iii) animals with intestinal inflammation induced by 3% of DSS, (iv) mice with intestinal inflammation induced by DSS and treated with BmooMP-alpha-I enzyme at the 50, 25, or 12.5 μg/animal/day dosages by intraperitoneal route. Clinical signs of colitis were observed daily for calculating the morbidity scores, cytokine measurements, and histological features. We observed that the animals treated with different doses of the enzyme presented a remarkable improvement of colitis signs, as confirmed by a significant increase of the intestine length in comparison to the DSS group. Also, no difference was observed between the groups treated with the enzyme or vehicle, as the colon length of these animals was slightly lower than that of the group of healthy animals, without induction of intestinal inflammation. The cytokine quantification in supernatants of intestinal tissue homogenates showed a significant reduction of 38% in IFN-gamma levels, when the animals were treated with 50 μg of the BmooMP-alpha-I compared to the animals receiving DSS only. A significant reduction of 39% in TNF levels was also observed in all doses of treatment with BmooMP-alpha-I, in addition to a significant reduction of 35% in the amount of IL-12p40. Histological examinations revealed that the BmooMP-alpha-I 50 μg treated group preserved colon architecture and goblet cells and reduced the ulcer area, when compared with DSS mice, which showed typical inflammatory changes in tissue architecture, such as ulceration, crypt dilation, loss of tissue architecture, and goblet cell depletion, accompanied by a significant cell infiltration. In conclusion, our results suggest that the improvement of clinical scores and histological findings related to BmooMP-alpha-I treatment in this experimental model could be attributed to the metalloprotease ability to modulate cytokine production locally at the inflamed intestine. These findings highlight the potential anti-inflammatory role and effectiveness of this enzyme as a therapeutic alternative in this type of immunopathological condition.
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- 2018
31. Inducible Nitric Oxide Synthase is required for parasite restriction and inflammatory modulation during Neospora caninum infection
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Lourenço Faria Costa, Vanessa dos Santos Miranda, Silas Silva Santana, Ana Cláudia Arantes Marques Pajuaba, Patrício da Silva Cardoso Barros, Eliézer Lucas Pires Ramos, Flávia Batista Ferreira, Caroline M. Mota, José Roberto Mineo, and Tiago W. P. Mineo
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Nitric Oxide Synthase Type II ,Inflammation ,Nitric Oxide ,Parasite load ,Microbiology ,Nitric oxide ,Proinflammatory cytokine ,Interferon-gamma ,Mice ,chemistry.chemical_compound ,Immune system ,Immunity ,parasitic diseases ,medicine ,Animals ,General Veterinary ,biology ,Coccidiosis ,Interleukin-12 Subunit p40 ,Macrophages ,Neospora ,General Medicine ,biology.organism_classification ,Neospora caninum ,Mice, Inbred C57BL ,Nitric oxide synthase ,chemistry ,biology.protein ,Parasitology ,medicine.symptom - Abstract
Neospora caninum infection is an important cause of neuromuscular disease in dogs and abortion in cattle, leading to significant economic losses in beef and dairy industries. The protective immunity against apicomplexan parasites, specifically Toxoplasma gondii and N. caninum, is typically achieved by inducing an IL-12-driven Th1 immune response. IL-12 stimulates IFN-γ production, which activates Inducible Nitric Oxide Synthase (iNOS) and promotes consequent Nitric Oxide (NO) synthesis, classically described as one of the main effector mechanisms for parasite elimination. Here, we aimed to evaluate the role played by iNOS during N. caninum infection. Our results show that N. caninum infection in C57BL/6 wild type (WT) mice induce NO production in vivo and in vitro. In agreement, iNOS deficient mice, as well as WT mice treated with iNOS inhibitor aminoguanidine, succumbed during acute infection with a dose lethal to 50 % of the WT mice, and presented significant increase in parasite load when submitted to sub-lethal infection protocols. Interestingly, the lack of control of parasite proliferation observed in iNOS-/- mice was associated with notable CNS inflammation and increased production of the main systemic proinflammatory cytokines (IL-12, IFN-γ, IL-6, TNF and IL-17A). Taken together, our findings show that iNOS plays an important role in restricting N. caninum replication, while also modulates the inflammatory process induced by the infection.
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- 2019
32. Annexin A1 peptide is able to induce an anti-parasitic effect in human placental explants infected by Toxoplasma gondii
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José Roberto Mineo, Ana Elizabete Silva, Eloisa Amália Vieira Ferro, Sonia Maria Oliani, Francesca Ietta, Luana Paulesu, A.O. Gomes, Caroline de Freitas Zanon, Jusciele Brogin Moreli, Marystela Fávero de Oliveira Cardoso, Universidade de São Paulo (USP), Faceres School of Medicine, Federal University of Triângulo Mineiro (UFTM), Universidade Estadual Paulista (Unesp), University of Siena, and Universidade Federal de Uberlândia (UFU)
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0301 basic medicine ,endocrine system ,Placenta ,Pregnancy Trimester, Third ,Anti-Inflammatory Agents ,Microbiology ,Dinoprostone ,Andrology ,03 medical and health sciences ,0302 clinical medicine ,Pregnancy ,parasitic diseases ,medicine ,Humans ,Prostaglandin E2 ,Receptors, Lipoxin ,Receptor ,Ac2-26peptide ,Annexin A1 ,Inflammation ,biology ,Antiparasitic Agents ,Toxoplasma gondii ,Toxoplasmosis ,Infectious Diseases ,biology.organism_classification ,medicine.disease ,beta-Galactosidase ,Antiparasitic agent ,Receptors, Formyl Peptide ,Ac2-26 peptide ,030104 developmental biology ,medicine.anatomical_structure ,Cross-Sectional Studies ,Cyclooxygenase 2 ,030220 oncology & carcinogenesis ,Immunohistochemistry ,Female ,Peptides ,Toxoplasma ,medicine.drug - Abstract
Made available in DSpace on 2018-12-11T17:21:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-10-01 Made available in DSpace on 2018-12-11T17:21:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-10-01 This study was conducted to investigate annexin A1 (ANXA1) functions in human placental explants infected with Toxoplasma gondii (T. gondii). We examined the first and third trimester placental explants infected with T. gondii (n = 7 placentas/group) to identify the number and location of parasites, ANXA1 protein, potential involvement of formyl peptide receptors (FPR1 and FPR2), and COX-2 expressions by immunohistochemistry. Treatments with Ac2-26 mimetic peptide of ANXA1 were performed to verify the parasitism rate (β-galactosidase assay), prostaglandin E2 levels (ELISA assay), and ANXA1, FPR1 and COX-2 expression in third trimester placentas. Placental explants of third trimester expressed less ANXA1 and were more permissive to T. gondii infection than first trimester placentas that expressed more ANXA1. Ac2-26 treatment increases endogenous ANXA1 and decreases parasitism rate, COX-2, and prostaglandin E2 levels. Altogether, these data provide further insight into the anti-parasitic and anti-inflammatory effects of ANXA1 in placentas infected with T. gondii. Post-Graduation in Structural and Functional Biology Federal University of São Paulo (UNIFESP) Faceres School of Medicine Department of Structural Biology Institute of Biological and Natural Sciences Federal University of Triângulo Mineiro (UFTM) Department of Biology Instituto de Biociências Letras e Ciências Exatas São Paulo State University (UNESP) Department of Life Sciences University of Siena Department of Immunology Microbiology and Parasitology Federal University of Uberlândia (UFU) Department of Histology and Embryology Federal University of Uberlândia (UFU) Department of Biology Instituto de Biociências Letras e Ciências Exatas São Paulo State University (UNESP)
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- 2018
33. Enrofloxacin and Toltrazuril Are Able to Reduce Toxoplasma gondii Growth in Human BeWo Trophoblastic Cells and Villous Explants from Human Third Trimester Pregnancy
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José Roberto Mineo, A.O. Gomes, Ariane S. Pereira, Iliana Claudia Balga Milián, Eloisa Amália Vieira Ferro, B.F. Barbosa, Rafaela José da Silva, Paolo Fiorenzani, Priscila Silva Franco, Neide M. Silva, Mayara Ribeiro, and Maria Célia dos Santos
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0301 basic medicine ,Microbiology (medical) ,placenta ,animal diseases ,medicine.medical_treatment ,030106 microbiology ,Immunology ,lcsh:QR1-502 ,Toxoplasma gondii ,Microbiology ,lcsh:Microbiology ,toltrazuril ,Andrology ,03 medical and health sciences ,chemistry.chemical_compound ,Sulfadiazine ,parasitic diseases ,Toltrazuril ,medicine ,Enrofloxacin ,treatment ,biology ,Trophoblast ,biology.organism_classification ,trophoblast ,Virology ,Neospora caninum ,030104 developmental biology ,Infectious Diseases ,Pyrimethamine ,medicine.anatomical_structure ,Cytokine ,chemistry ,embryonic structures ,enrofloxacin ,medicine.drug - Abstract
Classical treatment for congenital toxoplasmosis is based on combination of sulfadiazine and pyrimethamine plus folinic acid. Due to teratogenic effects and bone marrow suppression caused by pyrimethamine, the establishment of new therapeutic strategies is indispensable to minimize the side effects and improve the control of infection. Previous studies demonstrated that enrofloxacin and toltrazuril reduced the incidence of Neospora caninum and Toxoplasma gondii infection. The aim of the present study was to evaluate the efficacy of enrofloxacin and toltrazuril in the control of T. gondii infection in human trophoblast cells (BeWo line) and in human villous explants from the third trimester. BeWo cells and villous were treated with several concentrations of enrofloxacin, toltrazuril, sulfadiazine, pyrimethamine, or combination of sulfadiazine+pyrimethamine, and the cellular or tissue viability was verified. Next, BeWo cells were infected by T. gondii (2F1 clone or the ME49 strain), whereas villous samples were only infected by the 2F1 clone. Then, infected cells and villous were treated with all antibiotics and the T. gondii intracellular proliferation as well as the cytokine production were analyzed. Finally, we evaluated the direct effect of enrofloxacin and toltrazuril in tachyzoites to verify possible changes in parasite structure. Enrofloxacin and toltrazuril did not decrease the viability of cells and villous in lower concentrations. Both drugs were able to significantly reduce the parasite intracellular proliferation in BeWo cells and villous explants when compared to untreated conditions. Regardless of the T. gondii strain, BeWo cells infected and treated with enrofloxacin or toltrazuril induced high levels of IL-6 and MIF. In villous explants, enrofloxacin induced high MIF production. Finally, the drugs increased the number of unviable parasites and triggered damage to tachyzoite structure. Taken together, it can be concluded that enrofloxacin and toltrazuril are able to control T. gondii infection in BeWo cells and villous explants, probably by a direct action on the host cells and parasites, which leads to modifications of cytokine release and tachyzoite structure.
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- 2017
34. Proposed panel of diagnostic tools for accurate temporal classification of symptomatic T. gondii infection
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Deise A. O. Silva, Priscila Pinto Silva-dos-Santos, José Roberto Mineo, Geisa Baptista Barros, Jordana Grazziela Coelho-dos-Reis, Laurence Rodrigues do Amaral, José Carlos Serufo, Matheus de Souza Gomes, Elenice Moreira Lemos, Reynaldo Dietze, Eliana Zandonade, Olindo Assis Martins-Filho, and Ana C. A. M. Pajuaba
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0301 basic medicine ,Adult ,Male ,Time Factors ,Adolescent ,030106 microbiology ,Immunology ,Fluoroimmunoassay ,Antibodies, Protozoan ,Enzyme-Linked Immunosorbent Assay ,Diagnostic tools ,Serology ,03 medical and health sciences ,Young Adult ,Predictive Value of Tests ,medicine ,Immunology and Allergy ,Humans ,Serologic Tests ,Longitudinal Studies ,Prospective Studies ,Longitudinal cohort ,Child ,Fluorescent Antibody Technique, Indirect ,Aged ,Aged, 80 and over ,biology ,Toxoplasma gondii ,Igg subclasses ,Igg avidity ,Middle Aged ,medicine.disease ,biology.organism_classification ,Virology ,Toxoplasmosis ,Immunoglobulin A ,030104 developmental biology ,Immunoglobulin M ,ROC Curve ,Area Under Curve ,Immunoglobulin G ,Host-Pathogen Interactions ,Female ,Toxoplasma ,Biomarkers - Abstract
Serological tests available for the diagnosis of acute Toxoplasma gondii infection have limitations in establishing the temporal diagnosis of acute toxoplasmosis. The present analytical-descriptive investigation comprises of a prospective longitudinal cohort study to search for accurate biomarkers to distinguish acute, early and late convalescent T. gondii infection. Classic methods (immunofluorescence-IFA along with Enzyme-linked immunosorbent-ELISA and fluorescent-ELFA assays) for IgM, IgA, IgG and IgG avidity were employed in parallel with flow cytometry-based anti-fixed T. gondii tachyzoites serology (FC-AFTA-IgM, IgG, IgG avidity and IgG subclasses). The results reemphasized the limitations of IgM & IgG IFA, IgG ELFA, IgG & IgG subclasses FC as well as IgA ELISA biomarkers for the temporal diagnosis of acute toxoplasmosis. Receiver Operating-characteristics features (ROC-curves) were employed to adjust conventional cut-offs aiming at establishing a novel protocol to discriminate more accurately the different phases of toxoplasmosis. Conversely, IgM presented high diagnostic co-positivity for acute toxoplasmosis (97% for ELISA, 96% for ELFA and 95% for FC-AFTA) along with moderate co-negativity for detection of late convalescent toxoplasmosis (82%, 76% and 79%, respectively). IgG avidity (ELFA and FC-AFTA) outstand with the highest performance indices with 91% and 96% co-negativity for assessing acute toxoplasmosis and 91% and 98% co-positivity for late convalescent toxoplasmosis, respectively. Multivariate analysis generated a three-step algorithm comprising IgM ELFA screening followed by ELFA and FC-AFTA IgG avidity with high accuracy in discriminating acute from late convalescent infection. Together, these findings demonstrate the applicability of the proposed panel of diagnostic tools for accurate temporal classification of T. gondii infection.
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- 2017
35. Toxoplasma gondii antigen SAG2A differentially modulates IL-1β expression in resistant and susceptible murine peritoneal cells
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Thaise Anne Rocha Dos Santos, Carlos Priminho Pirovani, José Roberto Mineo, Jair P. Cunha-Junior, Edson Mario de Andrade, Juliano Oliveira Santana, Tiago W. P. Mineo, Jane Lima dos Santos, Tiago Antônio de Oliveira Mendes, and Jamilly Azevedo Leal-Sena
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0301 basic medicine ,Proteomics ,Cell ,Interleukin-1beta ,Macrophage polarization ,Protozoan Proteins ,Inflammation ,Antigens, Protozoan ,Applied Microbiology and Biotechnology ,Microbiology ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Immune system ,Antigen ,parasitic diseases ,Gene expression ,medicine ,Animals ,Humans ,Cells, Cultured ,Mice, Inbred BALB C ,biology ,Interleukin ,Toxoplasma gondii ,General Medicine ,biology.organism_classification ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,Macrophages, Peritoneal ,Female ,medicine.symptom ,Toxoplasma ,Toxoplasmosis ,030215 immunology ,Biotechnology - Abstract
The cell surface of Toxoplasma gondii is covered by antigens (SAGs) from the SRS family anchored by glycosylphosphatidylinositol (GPI) and includes antigens from the SAG2 family. Among these, the SAG2A surface antigen shows great potential in activating humoral responses and has been used in characterizing the acute phase of infection and in the serological diagnosis of toxoplasmosis. In this study, we aimed to evaluate rSAG2A-induced proteins in BALB/c and C57BL/c mice macrophages and to evaluate the phenotypic polarization induced in the process. We treated the peritoneal macrophages from mouse strains that were resistant or susceptible to T. gondii with rSAG2A to analyze their proteomic profile by mass spectrometry and systems biology. We also examined the gene expression of these cells by RT-qPCR using the phenotypic markers of M1 and M2 macrophages. Differences were observed in the expression of proteins involved in the inflammatory process in both resistant and susceptible cells, and macrophages were preferentially induced to obtain a pro-inflammatory immune response (M1) via the overexpression of IL-1β in mice susceptible to this parasite. These data suggest that the SAG2A antigen induces phenotypic and classical activation of macrophages in both resistant and susceptible strains of mice during the acute phase of the disease.
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- 2017
36. Rottlerin-mediated inhibition of Toxoplasma gondii growth in BeWo trophoblast-like cells
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Elena Daveri, Rafaela José da Silva, Anna Maria Avanzati, Roberta Romagnoli, Eloisa Amália Vieira Ferro, J.G. Oliveira, Emanuela Maioli, Luana Paulesu, Francesca Ietta, Laura Cresti, B.F. Barbosa, A.O. Gomes, and José Roberto Mineo
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0301 basic medicine ,Science ,Antiprotozoal Agents ,Article ,Cell Line ,03 medical and health sciences ,chemistry.chemical_compound ,parasitic diseases ,Autophagy ,Humans ,Benzopyrans ,Pathogen ,PI3K/AKT/mTOR pathway ,Multidisciplinary ,biology ,Intracellular parasite ,Acetophenones ,Toxoplasma gondii ,Translation (biology) ,biology.organism_classification ,Trophoblasts ,Cell biology ,030104 developmental biology ,chemistry ,Cell culture ,Protein Biosynthesis ,Medicine ,Toxoplasma ,Rottlerin - Abstract
Autophagy is a crucial and physiological process for cell survival from yeast to mammals, including protozoan parasites. Toxoplasma gondii, an intracellular parasite, typically exploits autophagic machinery of host cell; however host cell upregulates autophagy to combat the infection. Herein we tested the efficacy of Rottlerin, a natural polyphenol with autophagic promoting properties, against Toxoplasma infection on the chorioncarcinoma-derived cell line BeWo. We found that Rottlerin, at sub-toxic doses, induced morphological and biochemical alterations associated with autophagy and decreased Toxoplasma growth in infected cells. Although autophagy was synergically promoted by Toxoplasma infection in combination with Rottlerin treatment, the use of the autophagy inhibitor chloroquine revealed that Rottlerin anti-parasitic effect was largely autophagy-independent and likely mediated by the converging inhibitory effect of Rottlerin and Toxoplasma in host protein translation, mediated by mTOR inhibition and eIF2α phosphorylation. Both events, which on one hand could explain the additive effect on autophagy induction, on the other hand led to inhibition of protein synthesis, thereby depriving Toxoplasma of metabolically essential components for multiplication. We suggest that modulation of the competition between pathogen requirement and host cell defense might be an attractive, novel therapeutic approach against Toxoplasma infection and encourage the development of Rottlerin-based new therapeutic formulations.
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- 2017
37. Development of direct assays for Toxoplasma gondii and its use in genomic DNA sample
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Márcia Aires Rodrigues de Freitas, Lívia M. Alves, João M. Madurro, José Roberto Mineo, Vinícius de Rezende Rodovalho, Caroline M. Mota, Ana G. Brito-Madurro, Tiago W. P. Mineo, and Ana Cristina Honorato De Castro
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0301 basic medicine ,Guanine ,Clinical Biochemistry ,Pharmaceutical Science ,02 engineering and technology ,Analytical Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,parasitic diseases ,Drug Discovery ,Hydroxybenzoates ,Spectroscopy ,biology ,Chemistry ,Hybridization probe ,Toxoplasma gondii ,DNA ,Genomics ,021001 nanoscience & nanotechnology ,biology.organism_classification ,Molecular biology ,genomic DNA ,030104 developmental biology ,Colloidal gold ,Differential pulse voltammetry ,0210 nano-technology ,Biosensor ,Toxoplasma ,Toxoplasmosis - Abstract
This work describes an approach for the selection and detection of specific DNA probes related to Toxoplasma gondii, a protozoan parasite responsible for toxoplasmosis. The detection system was developed on graphite carbon electrode modified with poly(3-hydroxybenzoic acid) sensitized with ToxG1 probe. The hybridization of the specific genomic DNA related to T. gondii showed good response by direct detection of guanine residue oxidation using differential pulse voltammetry (DPV). The biosensor was able to distinguish both the complementary and non-complementary targets and detect up to 100ngμL-1 of the T. gondii genomic DNA. The hybridization (ToxG1: T. gondii genomic DNA) was confirmed by optical measurement. Optical assays using gold nanoparticles:ToxG1 probe showed a significant change in the absorbance peak in the presence of the T. gondii genomic DNA according to the electrochemical results. This novel biosensor shows potential as electrochemical transducer and was successfully applied in the biological sample.
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- 2017
38. Dectin-1 Compromises Innate Responses and Host Resistance against Neospora caninum Infection
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José Roberto Mineo, Eliézer Lucas Pires Ramos, Tiago W. P. Mineo, Fernanda Maria Santiago, Caroline M. Mota, Murilo V. Silva, Flávia Batista Ferreira França, and Arlindo Gomes de Macedo Júnior
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reactive oxygen species ,0301 basic medicine ,CLEC7A ,Innate immune system ,biology ,030106 microbiology ,Immunology ,Neospora caninum ,Toxoplasma gondii ,biology.organism_classification ,Parasite load ,macrophages ,Pathogenesis ,03 medical and health sciences ,030104 developmental biology ,Immunology and Allergy ,dendritic cells ,Signal transduction ,Receptor ,laminarin ,IL-12p40 - Abstract
Neospora caninum is an intracellular protozoan parasite that has drawn increasing interest due to its association with worldwide repetitive bovine abortions, which cause billionaire losses to the meat and dairy industries annually. Innate immunity plays an important role in infection control, and N. caninum activates the production of inflammatory mediators through Toll-like Receptors (TLR), Nod-Like Receptors (NLR) and Mitogen Activated Protein Kinase (MAPK) signaling pathways. Advances in the knowledge of initial host-parasite interactions are desirable for the design of control measures against the infection, obliterating its pathogenesis. In that sense, we here aimed to describe the role of the innate C-type lectin receptor (CLR) Dectin-1 during the infection by N. caninum. With that intent, we observed that the absence of Dectin-1, observed in genetically depleted (Dectin-1-/-) mice or competitively inhibited by an inert agonist (Laminarin, LAM), rescued 50% of the mice infected with lethal doses of N. caninum. Dectin-1-/- and LAM-treated mice also presented a reduction in the parasite load during acute and chronic phases, associated to decreased inflammatory scores in the central nervous system. Amongst all the cell phenotypes that migrated to the initial site of infection, dendritic cells and macrophages gained subpopulations with high Dectin-1 surface expression. The impairment of the receptor in these cells led to a decreased parasite burden, as well as augmented production of IL-12p40. We also found that Dectin-1+ cells produced less reactive oxygen species (ROS) at the initial site of the infection, while mice deficient in NAPDH oxidase isoform 2 (NOX2-/-) were not able to control parasite replication and produce IL-12p40, even upon LAM treatment. Interestingly, the absence of functional Dectin-1 did not alter the susceptibility of mice against closely related T. gondii. In conclusion, the gathered data suggests that Dectin-1 is involved in the parasite-induced downmodulation of ROS and other key molecules triggered for the control of N. caninum infection, and is a promising target for future development of protocols intended for intervention against neosporosis.
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- 2017
39. IL-17-Expressing CD4+and CD8+T Lymphocytes in Human Toxoplasmosis
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Marcos Vinicius da Silva, Neide M. Silva, César Gómez-Hernández, Virmondes Rodrigues Junior, Karine Rezende-Oliveira, Jéssica Líver Alves Silva, José Roberto Mineo, and Bethânea Crema Peghini
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Adult ,CD4-Positive T-Lymphocytes ,Article Subject ,medicine.medical_treatment ,T cell ,Immunology ,CD8-Positive T-Lymphocytes ,Biology ,Interleukin 21 ,Immunophenotyping ,lcsh:Pathology ,medicine ,Humans ,Cytotoxic T cell ,Interleukin-17 ,Cell Biology ,Middle Aged ,Cytokine ,medicine.anatomical_structure ,Leukocytes, Mononuclear ,Interleukin 12 ,Female ,Interleukin 17 ,Toxoplasmosis ,CD8 ,Research Article ,lcsh:RB1-214 - Abstract
This study aimed to measure the synthesis of Th1 and Th2 cytokines by mononuclear cells after culture with liveT. gondiiand identified Th17 (CD4+) and Tc17 (CD8+) cells in toxoplasma-seronegative and toxoplasma-seropositive parturient and nonpregnant women. Cytometric bead arrays were used to measure cytokine levels (IL-2, TNF-α, IFN-γ, IL-4, IL-5, and IL-10); immunophenotyping was used to characterize Th17 and Tc17 cells, and the cells were stained with antibodies against CD4+and CD8+T cells expressing IL-17. The addition of tachyzoites to cell cultures induced the synthesis of IL-5, IL-10, and TNF-αby cells from seronegative parturient women and of IL-5 and IL-10 by cells from seropositive, nonpregnant women. We observed a lower level of IL-17-expressing CD4+and CD8+T lymphocytes in cultures of cells from seronegative and seropositive parturient and nonpregnant women that were stimulated with tachyzoites, whereas analysis of the CD4+and CD8+T cell populations showed a higher level of CD4+T cells compared with CD8+T cells. These results suggest that the cytokine pattern and IL-17-expressing CD4+and CD8+T lymphocytes may have important roles in the inflammatory response toT. gondii, thus contributing to the maintenance of pregnancy and control of parasite invasion and replication.
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- 2014
40. Manual ilustrado de práticas laboratoriais em imunologia
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José Roberto Mineo, Maraísa Cristina Silva, Helena Maria Caleiro Acerbi Penha, and Paula Cristina Brígido
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- 2016
41. Dectin-1 Compromises Innate Responses and Host Resistance against
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Murilo Vieira, da Silva, Flávia Batista, Ferreira França, Caroline Martins, Mota, Arlindo Gomes, de Macedo Júnior, Eliézer Lucas Pires, Ramos, Fernanda Maria, Santiago, José Roberto, Mineo, and Tiago Wilson Patriarca, Mineo
- Subjects
reactive oxygen species ,CLEC7A ,Immunology ,Neospora caninum ,Toxoplasma gondii ,dendritic cells ,laminarin ,Original Research ,IL-12p40 ,macrophages - Abstract
Neospora caninum is an intracellular protozoan parasite that has drawn increasing interest due to its association with worldwide repetitive bovine abortions, which cause billionaire losses to the meat and dairy industries annually. Innate immunity plays an important role in infection control, and N. caninum activates the production of inflammatory mediators through toll-like receptors, NOD-like receptors, and mitogen-activated protein kinase signaling pathways. Advances in the knowledge of initial host–parasite interactions are desirable for the design of control measures against the infection, obliterating its pathogenesis. In that sense, we here aimed to describe the role of the innate C-type lectin receptor Dectin-1 during the infection by N. caninum. With that intent, we observed that the absence of Dectin-1, observed in genetically depleted (Dectin-1−/−) mice or competitively inhibited by an inert agonist [laminarin (LAM)], rescued 50% of the mice infected with lethal doses of N. caninum. Dectin-1−/− and LAM-treated mice also presented a reduction in the parasite load during acute and chronic phases, associated with decreased inflammatory scores in the central nervous system. Among all the cell phenotypes that migrated to the initial site of infection, dendritic cells and macrophages gained subpopulations with high Dectin-1 surface expression. The impairment of the receptor in these cells led to a decreased parasite burden, as well as augmented production of IL-12p40. We also found that Dectin-1+ cells produced less reactive oxygen species (ROS) at the initial site of the infection, while mice deficient in NADPH oxidase isoform 2 (NOX2−/−) were not able to control parasite replication and produce IL-12p40, even upon LAM treatment. Interestingly, the absence of functional Dectin-1 did not alter the susceptibility of mice against closely related Toxoplasma gondii. In conclusion, the gathered data suggest that Dectin-1 is involved in the parasite-induced downmodulation of ROS, and other key molecules triggered for the control of N. caninum infection and are a promising target for future development of protocols intended for intervention against neosporosis.
- Published
- 2016
42. Lectins from Synadenium carinatum (ScLL) and Artocarpus heterophyllus (ArtinM) Are Able to Induce Beneficial Immunomodulatory Effects in a Murine Model for Treatment of Toxoplasma gondii Infection
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Leandro Peixoto Ferreira Souza, Eliézer Lucas Pires Ramos, Silas Silva Santana, Murilo Vieira Silva, Fernanda Maria Santiago, Tiago WP Mineo, Maria Cristina Roque-Barreira, and José Roberto Mineo
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ScLL ,0301 basic medicine ,Microbiology (medical) ,medicine.medical_treatment ,Immunology ,lcsh:QR1-502 ,Toxoplasma gondii ,Therapeutics ,Microbiology ,Parasite load ,lcsh:Microbiology ,therapeutic agents ,ArtinM ,03 medical and health sciences ,Immune system ,Sulfadiazine ,medicine ,Cytotoxic T cell ,biology ,biology.organism_classification ,medicine.disease ,lectins ,Toxoplasmosis ,030104 developmental biology ,Infectious Diseases ,Cytokine ,Pyrimethamine ,medicine.drug - Abstract
Infection by Toxoplasma gondii affects around one-third of world population and the treatment for patients presenting toxoplasmosis clinically manifested disease is mainly based by a combination of sulfadiazine, pyrimethamine, and folinic acid. However, this therapeutic protocol is significantly toxic, causing relevant dose-related bone marrow damage. Thus, it is necessary to improve new approaches to investigate the usefulness of more effective and non-toxic agents for treatment of patients with toxoplasmosis. It has been described that lectins from plants can control parasite infections, when used as immunological adjuvants in vaccination procedures. This type of lectins, such as ArtinM and ScLL is able to induce immunostimulatory activities, including efficient immune response against parasites. The present study aimed to evaluate the potential immunostimulatory effect of ScLL and ArtinM for treatment of T. gondii infection during acute phase, considering that there is no study in the literature accomplishing this issue. For this purpose, bone marrow-derived macrophages (BMDMs) were treated with different concentrations from each lectin to determine the maximum concentration without or with lowest cytotoxic effect. After, it was also measured the cytokine levels produced by these cells when stimulated by the selected concentrations of lectins. We found that ScLL showed high capacity to induce of pro-inflammatory cytokine production, while ArtinM was able to induce especially an anti-inflammatory cytokines production. Furthermore, both lectins were able to increase NO levels. Next, we evaluated the treatment effect of ScLL and ArtinM in C57BL/6 mice infected by ME49 strain from T. gondii. The animals were infected and treated with ScLL, ArtinM, ArtinM plus ScLL, or sulfadiazine, and the following parameters analyzed: Cytokines production, brain parasite burden and survival rates. Our results demonstrated that the ScLL or ScLL plus ArtinM treatment induced production of pro-inflammatory and anti-inflammatory cytokines, showing differential but complementary profiles. Moreover, when compared with non-treated mice, the parasite burden was significantly lower and survival rates higher in mice treated with ScLL or ScLL plus ArtinM, similarly with sulfadiazine treatment. In conclusion, the results demonstrated the suitable potential immunotherapeutic effect of ScLL and ArtinM lectins to control acute toxoplasmosis in this experimental murine model.
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- 2016
43. Si-Accumulation In Artemisia annua Glandular Trichomes Increases Artemisinin Concentration, but Does Not Interfere In the Impairment of Toxoplasma gondii Growth
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Deise A. O. Silva, Xavier Simonnet, Lilian Aparecida de Oliveira, Mônica Lanzoni Rossi, Cristina Rostkowska, Caroline M. Mota, Regina Maria Quintão Lana, Tiago W. P. Mineo, Fernanda Maria Santiago, José Roberto Mineo, Gaspar Henrique Korndörfer, Taísa Carrijo de Oliveira, and Neusa L. Nogueira
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0301 basic medicine ,biology ,Magnesium silicate ,030106 microbiology ,Artemisia annua ,food and beverages ,chemistry.chemical_element ,Toxoplasma gondii ,Plant Science ,Calcium ,Micronutrient ,biology.organism_classification ,Trichome ,03 medical and health sciences ,Horticulture ,030104 developmental biology ,chemistry ,parasitic diseases ,Botany ,medicine ,Artemisinin ,Medicinal plants ,medicine.drug - Abstract
Artemisia annua is used as a source of artemisinin, a potent therapeutic agent used for the treatment of infectious diseases, chiefly malaria. However, the low concentration (from 0.01 to 1.4% of dried leaf matter) of artemisinin in the plant obtained with the traditional cropping system makes it a relatively expensive drug, especially in developing countries. Considering that artemisinin and silicon (Si) are both stored in A. annua glandular trichomes, and that Si accumulation has never been investigated, this study aimed to look into Si effects on A. annua trichome artemisinin concentration, and whether leaf infusion from Si-treated A. annua plants is able to control Toxoplasma gondii growth. T. gondii is the etiologic agent of toxoplasmosis, a zoonotic parasitic disease whose traditional treatment shows significant side effects. The experimental design consisted of A. annua seedlings randomly planted in soil treated with different doses of calcium/magnesium silicate (0, 200, 400, 800, and 1600 kg ha-1). Analysis of foliar macronutrients showed significant increases of nitrogen content only at the highest dose of silicate. Foliar micronutrients, Si concentrations, and plant height were not affected by any of the silicate doses. However, the dose of 400 kg ha-1 of silicate increased the trichome size, which in turn raised artemisinin concentration in leaves and the infusion. In contrast, the 800 and 1600 kg ha-1 doses dramatically decreased artemisinin concentration. HeLa cell treatment with the infusion of A. annua grown in soil treated with 400 kg ha-1 of silicate decreased parasite proliferation in a dose-dependent manner when the treatment was carried out after or along with T. gondii infection. However, this effect was similar to A. annua grown in soil without silicate treatment. Thus, it can be concluded that, even though Si applied to the soil at 400 kg ha-1 has a positive effect on the A. annua glandular trichome size and the artemisinin concentration, this outcome cannot be directly associated with the efficiency of A. annua infusion on T. gondii growth, suggesting that other components from A. annua leaves could be acting in synergy with artemisinin.
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- 2016
44. Toxoplasma gondii-Derived Synthetic Peptides Containing B- and T-Cell Epitopes from GRA2 Protein Are Able to Enhance Mice Survival in a Model of Experimental Toxoplasmosis
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Tiago Wp Mineo, Carlos Priminho Pirovani, Murilo Veira Silva, Fabiana de Almeida Araújo Santos, Fernanda Maria Santiago, Arlindo Gomes de Macêdo, Luciana Machado Bastos, Eliézer Lucas Pires Ramos, Luiz Ricardo Goulart, and José Roberto Mineo
- Subjects
Protozoan Vaccines ,0301 basic medicine ,Phage display ,Protein Conformation ,Protozoan Proteins ,lcsh:QR1-502 ,Antibodies, Protozoan ,Epitopes, T-Lymphocyte ,lcsh:Microbiology ,Epitope ,Mice ,Fluorescent Antibody Technique, Indirect ,Original Research ,biology ,Antibodies, Monoclonal ,B- and T-cell epitopes ,Isotype ,Survival Rate ,Treatment Outcome ,T-cell epitope ,Infectious Diseases ,GRA2 ,Cytokines ,Epitopes, B-Lymphocyte ,synthetic peptides ,Female ,Toxoplasma ,Toxoplasmosis ,Microbiology (medical) ,medicine.drug_class ,030106 microbiology ,Immunology ,Toxoplasma gondii ,Antigens, Protozoan ,Monoclonal antibody ,Microbiology ,03 medical and health sciences ,Adjuvants, Immunologic ,medicine ,Animals ,Amino Acid Sequence ,Rhoptry ,Interleukins ,B-cell epitope ,biology.organism_classification ,medicine.disease ,Virology ,Immunity, Humoral ,Mice, Inbred C57BL ,Models, Structural ,Disease Models, Animal ,030104 developmental biology ,monoclonal antibody ,Dense granule ,Peptides - Abstract
Toxoplasmosis is a zoonosis distributed all over the world, which the etiologic agent is an intracellular protozoan parasite, Toxoplasma gondii. This disease may cause abortions and severe diseases in many warm-blood hosts, including humans, particularly the immunocompromised patients. The parasite specialized secretory organelles, as micronemes, rhoptries and dense granules, are critical for the successful parasitism. The dense granule protein 2 (GRA2) is a parasite immunogenic protein secreted during infections and previous studies have been shown that this parasite component is crucial for the formation of intravacuolar membranous nanotubular network (MNN), as well as for secretion into the vacuole and spatial organization of the parasites within the vacuole. In the present study, we produced a monoclonal antibody to GRA2 (C3C5 mAb, isotype IgG2b), mapped the immunodominant epitope of the protein by phage display and built GRA2 synthetic epitopes to evaluate their ability to protect mice in a model of experimental infection. Our results showed that synthetic peptides for B- and T-cell epitopes are able to improve survival of immunized animals. In contrast with non-immunized animals, the immunized mice with both B- and T-cell epitopes had a better balance of cytokines and demonstrated higher levels of IL-10, IL-4 and IL-17 production, though similar levels of TNF-alpha and IL-6 were observed. The immunization with both B- and T-cell epitopes resulted in survival rate higher than 85% of the challenged mice. Overall, these results demonstrate that immunization with synthetic epitopes for both B- and T-cells from GRA2 protein can be more effective to protect against infection by T. gondii.
- Published
- 2016
45. Cytokines and chemokines production by mononuclear cells from parturient women after stimulation with live Toxoplasma gondii
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Karine Rezende-Oliveira, José Roberto Mineo, Neide M. Silva, and V. Rodrigues Junior
- Subjects
Chemokine ,Toxoplasma gondii ,Biology ,Peripheral blood mononuclear cell ,CCL5 ,Toxoplasmosis, Congenital ,Proinflammatory cytokine ,Immune system ,Pregnancy ,parasitic diseases ,Obstetrics and Gynaecology ,medicine ,Immune Tolerance ,Humans ,Immune response ,Chemokine CCL5 ,Cells, Cultured ,Chemokine CCL2 ,Interleukin-6 ,Tumor Necrosis Factor-alpha ,Intracellular parasite ,Parturition ,Obstetrics and Gynecology ,biology.organism_classification ,medicine.disease ,Interleukin-12 ,Toxoplasmosis ,Interleukin-10 ,Reproductive Medicine ,Immunology ,biology.protein ,Leukocytes, Mononuclear ,Cytokines ,Female ,Chemokines ,Toxoplasma ,Developmental Biology - Abstract
Toxoplasma gondii is an obligate intracellular parasite that can cause variable clinical symptoms or can even be asymptomatic in immunocompetent individuals. More severe symptoms are observed in immunocompromised patients and congenital transmission of the parasite has been reported. The objective of this study was to evaluate the response of peripheral blood mononuclear cells (PBMC) in parturient and non-pregnant women exposed to live tachyzoites of T. gondii strain RH or ME49. PBMC were isolated from parturient and non-pregnant women with negative or positive serology for toxoplasmosis and cultured with live tachyzoites of the two T. gondii strains for 24 h. Next, the cell culture supernatants were collected and levels of CCL2, CCL5, IL-6, IL-10, IL-12, and TNF-α produced by PBMC after tachyzoite exposure were measured. Live tachyzoite forms of T. gondii significantly inhibited the synthesis of CCL2 in seropositive parturient women, whereas a stimulatory effect on CCL5 was observed in seronegative parturient women. Cells from T. gondii-seronegative non-pregnant women produced significantly higher levels of TNF-α and IL-12, demonstrating the proinflammatory profile induced by the presence of the parasite in culture. The results suggest that the immunomodulation seen during pregnancy contributes to the development of an environment that facilitates escape of the parasite from the immune response.
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- 2012
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46. Analysis of IgG subclasses (IgG1 and IgG3) to recombinant SAG2A protein from Toxoplasma gondii in sequential serum samples from patients with toxoplasmosis
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Reynaldo Dietze, Jair P. Cunha-Junior, Carlos Priminho Pirovani, Geisa Baptista Barros, Letícia de Almeida Leão Vaz, Deise A. O. Silva, José Roberto Mineo, Elenice Moreira Lemos, and Silas Silva Santana
- Subjects
Adult ,Male ,Time Factors ,Adolescent ,Immunology ,Protozoan Proteins ,Toxoplasma gondii ,Antigens, Protozoan ,law.invention ,Serology ,Young Adult ,Immune system ,Antigen ,law ,parasitic diseases ,medicine ,Humans ,Immunology and Allergy ,IgG subclasses ,Child ,Aged ,Aged, 80 and over ,biology ,Middle Aged ,biology.organism_classification ,medicine.disease ,Recombinant Proteins ,Toxoplasmosis ,Immunoglobulin A ,Kinetics ,Immunoglobulin M ,Immunoglobulin G ,Humoral immunity ,Recombinant DNA ,biology.protein ,Female ,Recombinant protein SAG2A ,Antibody ,Toxoplasma ,Serological diagnosis - Abstract
The kinetics of the humoral immune response was evaluated using the recombinant SAG2A protein comparatively to soluble Toxoplasma antigen (STAg) by ELISA in sequential serum samples of patients with toxoplasmosis up to 12 months of illness onset. The follow up of IgM and IgA levels to STAg showed a gradual decrease, with the majority of patients (88%) seropositive for IgM up to 12 months of infection, whereas IgA seropositivity was relatively low (78%) compared to IgM (100%) in the first 3 months of infection. The follow up of IgG and IgG1 antibodies showed a similar increasing profile for both SAG2A and STAg, with slightly higher seropositivity for STAg. The kinetics of IgG3 to STAg was similar to that of IgG1, contrasting with the kinetics of IgG3 to SAG2A that showed high levels up to 6 months of infection, with continuous decreasing over the time. Higher IgG3 seropositivity to SAG2A than STAg was also observed in the initial phases of infection. A higher IgG3/IgG1 ratio for SAG2A than STAg was detected in the first 3 months of infection, with decreasing profile over the time. The associations of IgG3/IgG1 ratio>1.0 with positive IgM or IgA antibodies were predominantly found in the first 3 months of infection, whereas associations of IgG3/IgG1 ratio
- Published
- 2012
47. Azithromycin and spiramycin induce anti-inflammatory response in human trophoblastic (BeWo) cells infected by Toxoplasma gondii but are able to control infection
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Deise A. O. Silva, Priscila Silva Franco, M.B. Angeloni, B.F. Barbosa, Eloisa Amália Vieira Ferro, Andréa Teixeira-Carvalho, Neide M. Silva, A.O. Gomes, José Roberto Mineo, and Olindo Assis Martins-Filho
- Subjects
Cell Survival ,Anti-Inflammatory Agents ,Toxoplasma gondii ,Inflammation ,Azithromycin ,Cell Line ,Microbiology ,Mice ,Pregnancy ,Obstetrics and Gynaecology ,Spiramycin ,parasitic diseases ,BeWo cells ,medicine ,Animals ,Humans ,Viability assay ,biology ,Choriocarcinoma ,Obstetrics and Gynecology ,medicine.disease ,biology.organism_classification ,Toxoplasmosis ,Trophoblasts ,Reproductive Medicine ,Cell culture ,embryonic structures ,Immunology ,Cytokines ,Female ,Macrophage migration inhibitory factor ,medicine.symptom ,Toxoplasma ,Developmental Biology ,medicine.drug - Abstract
Toxoplasma gondii is an important pathogen which may cause fetal infection if primary infection. Our previous studies have used human choriocarcinoma trophoblastic cells (BeWo cell line) as experimental model of T. gondii infection involving placental microenvironment. This study aimed to examine the effects of azithromycin and spiramycin against T. gondii infection in BeWo cells. Cells were treated with different concentrations of the macrolide antibiotics and analyzed first for cell viability using thiazolyl blue tetrazole (MTT) assay. As cell viability was significantly decreased with drug concentrations higher than 400 μg/mL, the concentration range used in further experiments was from 50 to 400 μg/mL. The number of infected cells and intracellular replication of T. gondii decreased after treatment with each drug. The infection induced up-regulation of the macrophage migration inhibitory factor (MIF), which was also enhanced in infected cells after treatment with azithromycin, but not with spiramycin. Analysis of the cytokine profile showed increase TNF-α, IL-10 and IL-4 production, but decreased IFN-γ levels, were detected in infected cells and treated with each drug. In conclusion, treatment of human trophoblastic BeWo cells with with azithromycin or spiramycin is able to control the infection and replication of T. gondii. In addition, treatment with these macrolides, especially with azityromycin induces an anti-inflammatory response and high MIF production, which can be important for the establishment and maintenance of a viable pregnancy during T. gondii infection.
- Published
- 2011
48. ArtinM, a d-mannose-binding lectin from Artocarpus integrifolia, plays a potent adjuvant and immunostimulatory role in immunization against Neospora caninum
- Author
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José Roberto Mineo, Julianne V. Carvalho, Caroline M. Mota, Tiago W. P. Mineo, Deise A. O. Silva, Fernanda Maria Santiago, Mariana R.D. Cardoso, Neide M. Silva, Dâmaso P. Ribeiro, Maria Cristina Roque-Barreira, and Ester C. B. Araújo
- Subjects
Antibodies, Protozoan ,Neospora caninum ,Parasite load ,Parasite Load ,Immunoglobulin G ,Mice ,ArtinM ,Immune system ,Neospora ,Adjuvants, Immunologic ,Antigen ,Jacalin ,Lectins ,Immunology and Microbiology(all) ,Animals ,General Veterinary ,General Immunology and Microbiology ,biology ,Coccidiosis ,Immunogenicity ,Public Health, Environmental and Occupational Health ,Brain ,biology.organism_classification ,veterinary(all) ,Mice, Inbred C57BL ,Mannose-Binding Lectins ,Infectious Diseases ,Immunology ,biology.protein ,Cytokines ,Molecular Medicine ,Female ,Immunization ,Plant Lectins ,Artocarpus ,Spleen - Abstract
ArtinM and Jacalin (JAC) are lectins from the jackfruit (Artocarpus integrifolia) that have important role in modulation of immune responses to pathogens. Neospora caninum is an Apicomplexa parasite that causes neuromuscular disease in dogs and reproductive disorders in cattle, with economic impact on the livestock industry. Hence, we evaluated the adjuvant effect of ArtinM and JAC in immunization of mice against neosporosis. Six C57BL/6 mouse groups were subcutaneously immunized three times at 2-week intervals with Neospora lysate antigen (NLA) associated with lectins (NLA+ArtinM and NLA+JAC), NLA, ArtinM and JAC alone, and PBS (infection control). Animals were challenged with lethal dose of Nc-1 isolate and evaluated for morbidity, mortality, specific antibody response, cytokine production by spleen cells, brain parasite burden and inflammation. Our results demonstrated that ArtinM was able to increase NLA immunogenicity, inducing the highest levels of specific total IgG and IgG2a/IgG1 ratio, ex vivo Th1 cytokine production, increased survival, the lowest brain parasite burden, along with the highest inflammation scores. In contrast, NLA+JAC immunized group showed intermediate survival, the highest brain parasite burden and the lowest inflammation scores. In conclusion, ArtinM presents stronger immunostimulatory and adjuvant effect than Jacalin in immunization of mice against neosporosis, by inducing a protective Th1-biased pro-inflammatory immune response and higher protection after parasite challenge.
- Published
- 2011
49. Bothrops pirajai snake venom L-amino acid oxidase: in vitro effects on infection of Toxoplasma gondii in human foreskin fibroblasts
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Lívia M. Alves, Deise A. O. Silva, Veridiana M. Rodrigues, Luiz Fernando Moreira Izidoro, and José Roberto Mineo
- Subjects
Oxidase test ,human fibroblasts ,biology ,Bothrops pirajai ,lcsh:RS1-441 ,Toxoplasma gondii ,L-amino-acid oxidase ,biology.organism_classification ,In vitro ,Microbiology ,lcsh:Pharmacy and materia medica ,Foreskin ,medicine.anatomical_structure ,L-amino acid oxidase ,Snake venom ,Toxicity ,medicine ,Cytotoxic T cell ,General Pharmacology, Toxicology and Pharmaceutics ,snake venom - Abstract
The effect of an L-amino acid oxidase isolated from Bothrops pirajai snake venom (BpirLAAO-I) was investigated on infection of Toxoplasma gondii in human foreskin fibroblasts (HFF). The cytotoxic activity of BpirLAAO-I on HFF cells showed a dose-dependent toxicity with median cytotoxic dose (TD50) of 11.8 µg/mL. BpirLAAO-I induced considerable dose-dependent decrease in the T. gondii infection index under two different conditions, treatment of tachyzoites before infection or treatment of HFF cells after infection. A maximal inhibition of infection (56%) was found for treatment before infection, with a median inhibitory dose (ID50) at 1.83 µg/mL and selectivity index (SI) at 6.45. For treatment after infection, it was observed a maximal inhibition of infection at 65%, ID50 of 1.20 µg/mL and SI of 9.83. The treatment before infection was also effective to reduce intracellular parasitism up to 62%, although presenting higher values of ID50 (3.14 µg/mL) and lower values of SI (3.76). However, treatment after infection was not effective, suggesting that the enzyme seems to have no effect on the parasite intracellular replication for this condition. In conclusion, BpirLAAO-I was more effective to inhibit the infection of neighboring cells and consequently parasite dissemination than primary infection and parasite replication. Thus, the effect of BpirLAAO-I described herein could be taken into account for the development of new synthetic anti-parasite therapeutic agents.
- Published
- 2011
50. Differential susceptibility of human trophoblastic (BeWo) and uterine cervical (HeLa) cells to Neospora caninum infection
- Author
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Caroline M. Mota, B.F. Barbosa, Tiago W. P. Mineo, Julianne V. Carvalho, Eloisa Amália Vieira Ferro, Deise A. O. Silva, Mariana R.D. Cardoso, C M O S Alves, José Roberto Mineo, and Neide M. Silva
- Subjects
Cattle Diseases ,Cervix Uteri ,Microbiology ,HeLa ,Neospora ,Antigen ,parasitic diseases ,Animals ,Humans ,Viability assay ,Innate immune system ,biology ,Coccidiosis ,Cell growth ,biology.organism_classification ,Virology ,Neospora caninum ,Trophoblasts ,Infectious Diseases ,Cell culture ,embryonic structures ,Cytokines ,Cattle ,Female ,Parasitology ,Disease Susceptibility ,HeLa Cells - Abstract
Neospora caninum is an apicomplexan parasite, closely related to Toxoplasma gondii, and causes abortion and congenital neosporosis in cattle worldwide. Trophoblast cells act in mechanisms of innate immune defense at the fetal-maternal interface and no data are available about the interaction of Neospora with human trophoblasts. Thus, this study aimed to verify the susceptibility of human trophoblastic (BeWo) compared with uterine cervical (HeLa) cell lines to N. caninum. BeWo and HeLa cells were infected with different parasite:cell ratios of N. caninum tachyzoites and analyzed at different times after infection for cell viability using thiazolyl blue tetrazole and lactate dehydrogenase assays. Both cell lines were also evaluated for cytokine production and parasite infection/replication assays when pre-treated or not with Neospora lysate antigen (NLA) or human recombinant IFN-γ. Cell viability was increased up to 48 h of infection in both types of cells, suggesting that infection could inhibit early cell death and/or induce cell proliferation. Neospora infection induced up-regulation of the macrophage migration inhibitory factor (MIF), mainly in HeLa cells, which was enhanced by cell pre-treatment by NLA or IFN-γ. Conversely, parasite infection induced down-regulation of the transforming growth factor (TGF-β), mostly in BeWo cells, which was decreased with NLA or IFN-γ pre-treatment. HeLa cells were more susceptible to Neospora infection than BeWo cells and IFN-γ pre-treatment resulted in reduced infection indices in both cell lines. Control of parasite growth was mediated by IFN-γ through an indoleamine-2,3-dioxygenase-dependent mechanism in HeLa cells alone. Based on these results, we concluded that BeWo and HeLa cells are readily infected by N. caninum, although presenting differences in susceptibility to infection, cytokine production and cell viability. Thus, these host cells can be considered in comparative approaches to understand strategies used by N. caninum to survive at the maternal-fetal interface.
- Published
- 2010
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