68 results on '"Fernandes ER"'
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2. Design and analysis of shell and tube heat exchanger
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Fernandes Erica Jacqueline and Krishanmurthy Sachidananda Hassan
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shell and tube exchanger ,energy ,baffle ,steady state ,Industrial engineering. Management engineering ,T55.4-60.8 ,Industrial directories ,T11.95-12.5 - Abstract
The demand for consumption of energy in industries has made designers to build efficient heat transfer exchangers. One of the most used heat exchangers which supports this is the shell and tube heat exchangers which are built for effective heat transfer. These heat exchangers are widely utilized in the HVAC industries especially in chiller plants due to their large surface for heat transfer. So, design of these chillers is influenced by the selection of material. This research paper discusses the design and analysis of shell and tube heat exchangers by considering different material and their ability to transfer heat from the surface. So, baffles play an important role to analyze the performance of the heat exchangers and it is possible to improve their heat transfer capabilities. So, in this research paper baffle spacing and its effect on heat transfer has been analyzed using CFD analysis and compared these results with the theoretical analysis. The Design and modelling of the heat exchanger have been modelled using PTC Creo parametric and using ANSYS Fluent CFD analysis have been carried out considering copper, aluminum, and steel as the materials. From this analysis it can be stated that copper has performed well as compared to aluminum and steel by using minimum baffle spacing.
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- 2022
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3. Chronic colitis associated with HIV infection can be related to intraepithelial infection of the colon by CD8+ T lymphocytes.
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Fernandes ER, Pagliari C, Tuon FF, Junior HFD, Averbach M, and Duarte MIS
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Gastrointestinal complications in AIDS patients with diarrhoea are common clinical manifestations, frequently diagnosed by colonoscopy as non-specific colitis. We retrospectively study colon biopsies diagnosed as chronic colitis associated with HIV (CCH). Biopsies were sorted as patients with AIDS (serum CD4 <200 cell/mm3) but without any clear infectious process (n = 12) and patients without HIV infection (n = 24). There are low numbers of CD4+ T lymphocytes in lamina propria of AIDS patients, but CD8+ T populations in this area appear to be similar in all studied groups, regardless of HIV infection or laboratory evidence of a specific agent. We found the clear evidence of CD8+ T cells infiltration in colonic mucosa in HIV patients with microscopic colitis. An imbalance of lymphocyte subpopulations in the colon, both in the lamina propria and epithelium, could result in an intraepithelial CD8 infiltration, involved in the pathogenesis of CCH in AIDS patients. [ABSTRACT FROM AUTHOR]
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- 2008
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4. Fe (III) - Galactomannan Solid and Aqueous Complexes: Potentiometric, EPR Spectroscopy and Thermal Data
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Mercê Ana L. R., Fernandes Erika, Mangrich Antonio S., Sierakowski M. R., and Szpoganicz Bruno
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iron (III) ,potentiometric titration ,binding constants ,EPR spectroscopy ,metal ion coordination ,Chemistry ,QD1-999 - Abstract
Galactomannans can be employed in food industries to modify the final rheological properties of the products. Since they are not absorbed by the living organisms they can also be used in dietary foods. The equilibria involving the interactions of Fe(III) and galactomannans and arabinogalactan of several leguminous plants were characterized by potentiometric titrations and EPR spectroscopy. The log of the equilibrium constants for the formation of ML species, where M is the metal ion and L is the monomeric unit of the biopolymers, were 15.4, 14.1 and 18.5, for the galactomannans of C. fastuosa, L. leucocephala and S. macranthera, respectively. Log K values for protonated species (MHL) were 3.1, 3.3, and were not detected for the galactomannan of S. macranthera. The log K values for the formation of ML2 were 14.1, 13.3 and 15.2, respectively. Early formation of insoluble products in the equilibrium with arabinogalactan and Fe(III) prevented acquisition of reliable data. The solid complexes assays showed a great dipolar interaction between two Fe(III) ions in the inner structure of the biopolymer which increased as the degree of substitution of the galactomannan decreased, and also showed the resulting thermal stability. The complexes impart a new possibility of providing essential metal ions in dietary foods since decomplexation of the complexes can occur at different pH values existing in the human body.
- Published
- 2001
5. Evaluation of the complexes of galactomannan of Leucaena leucocephala and Co2+, Mn2+ , Ni2+ and Zn2+
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Mercê Ana Lucia R., Fernandes Erika, Mangrich Antonio S., and Sierakowski Maria Rita
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potentiometric titrations ,EPR spectroscopy ,TG-DSC ,metal complexes ,Chemistry ,QD1-999 - Abstract
The binding constants for the complexed species formed in aqueous solution between galactomannan of Leucaena leucocephala and the metal ions Co2+, Mn2+, Ni2+ and Zn2+ were determined by potentiometric titrations. The calculated values showed Ni2+ as the best Lewis acid towards the Lewis base -OH groups of the sugar monomers, with Zn2+ being the poorest. For all systems, a higher percentage of the complexed species was present near pH=7.0, although complexed species existed over a wide range of acidic and basic pH values. The isolated solid complexes were studied by TG-DSC thermal analysis and by EPR spectroscopy. The thermal profiles obtained showed higher thermal resistance to final degradation than the biopolymer alone for the complexed species ML having the smallest log K values. The EPR spectra confirmed the complexation of the metal ions via the Lewis base deprotonated hydroxyl groups (-O) and showed that the distances between metal ions in the complexed biopolymer structure depend on the nature of the metal ion. The ability of galactomannans to complex a variety of metal ions in their web like structure and the resistance to high temperatures and a wide range of pH values of these complexes open new perspectives in possible industrial uses whenever these properties are required, such as in bioremediation of waste waters and in the application of slow-release fertilizers.
- Published
- 2000
6. Analysis of the antigenic and immunogenic properties of the native rabies virus glycoprotein purified by Lens culinaris lectin affinity chromatography.
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da Silva CP, Queiroz TGA, Nogi KI, Katz ISS, Guedes F, Fernandes ER, Silva KR, and Silva SR
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Rabies virus glycoprotein (RABV-G) is responsible for the recognition of specific cell surface receptors and induces the production of neutralizing antibodies (VNA). Since RABV-G is a glycoprotein, this work aimed to evaluate Lens culinaris (LCA) chromatography as a simple and effective purification method. The purity and identification of the protein obtained were analyzed by SDS-PAGE, ELISA and lectin-binding assay. The antigenic properties of the purified RABV-G were evaluated by direct ELISA using human serum samples from individuals who had received rabies pre-exposure vaccination. For the immunogenicity study, Swiss Webster mice were immunized with purified RABV-G and the specific antibodies were measured by direct ELISA and RFFIT. As results, it was observed that the purified protein reveled a molecular mass of 55 kDa and the presence of carbohydrate; additionally, it was recognized by anti-rabies virus glycoprotein monoclonal antibody. Purified RABV-G induced high VNA titers (>50.0 IU/ml) in vivo, as detected by RFFIT, as well as RABV-G specific IgG1 (0.8 mean OD±SD) and IgG2a (0.3 mean OD±SD) antibodies, with a predominance of IgG1 (p< 0.001). In addition, it was observed that RABV-G was efficient in selectively detecting anti- RABV-G IgG in the sera of vaccinated individuals compared to the negative control. Therefore, LCA chromatography was efficient in preserving the native properties of RABV-G that are essential in inducing an adequate humoral immune response. In addition, the purified RABV-G presented analytical potential as an ELISA reagent., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Conflict of interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)
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- 2024
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7. Seroepidemiological survey to cell culture rabies vaccines (CCRV) in Brazil.
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da Silva RI, Chaves LB, Dos Ramos Silva S, Katz ISS, Fernandes ER, Neto RC, Padovani CR, Modolo JR, Soares Magalhaes RJ, Crompton H, and Victoria C
- Abstract
Rabies is a contagious viral disease that can be easily transmitted by the saliva and brain/nervous system tissues of the infected animals, causing severe and fatal encephalitis in both animals and humans. Vaccination campaigns are crucial to combat and prevent rabies's spread in dogs and humans. The Modified Fuenzalida & Palicios vaccines have been widely used since the 70s and have proven effective in producing a solid serological response. Since 2008, the Brazilian Ministry of Health has introduced a Cell Culture Rabies Vaccine (CCRV) for all dog mass vaccination campaigns in Brazil. However, to date, there is limited evidence on the immunologic response of dogs to this type of vaccine in field conditions. The present study evaluated the serological response in dogs vaccinated with CCRV from blood samples of 724 dogs using the Simplified Fluorescence Inhibition Microtest - SFIMT. Dogs with a titer equal to 0.5 IU/mL or above were considered seropositive. The results revealed that 59.12% (428/724) of all dogs tested and 48.49% (32/66) of primo-vaccinated animals were seropositive. The percentage of seronegative animals was higher than seropositive for animals that received a single dose during their life ( p < 0.05). The opposite was observed in animals with five or more doses. The results of this study demonstrated that the CCRV vaccines elicit a satisfactory immunological response in field conditions and can constitute an essential population-level preventive strategy as part of annual canine rabies vaccination campaigns. Although its effectiveness has been studied, there is limited evidence of its immunological response in dogs under field conditions. This paper evaluates the serological response to CCRV in dogs vaccinated during mass vaccination campaigns from 2012 to 2017., Competing Interests: The author(s) declares no conflict of interest in this work., (© 2024 Published by Elsevier B.V.)
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- 2024
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8. Heterologous DNA Prime- Subunit Protein Boost with Chikungunya Virus E2 Induces Neutralizing Antibodies and Cellular-Mediated Immunity.
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Coirada FC, Fernandes ER, Mello LR, Schuch V, Soares Campos G, Braconi CT, Boscardin SB, and Santoro Rosa D
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- Animals, Mice, Antibodies, Neutralizing, CD8-Positive T-Lymphocytes, Antibodies, Viral, Immunity, Cellular, DNA, Chikungunya virus genetics, Viral Vaccines, Vaccines, DNA
- Abstract
Chikungunya virus (CHIKV) has become a significant public health concern due to the increasing number of outbreaks worldwide and the associated comorbidities. Despite substantial efforts, there is no specific treatment or licensed vaccine against CHIKV to date. The E2 glycoprotein of CHIKV is a promising vaccine candidate as it is a major target of neutralizing antibodies during infection. In this study, we evaluated the immunogenicity of two DNA vaccines (a non-targeted and a dendritic cell-targeted vaccine) encoding a consensus sequence of E2
CHIKV and a recombinant protein (E2*CHIKV ). Mice were immunized with different homologous and heterologous DNAprime-E2* protein boost strategies, and the specific humoral and cellular immune responses were accessed. We found that mice immunized with heterologous non-targeted DNA prime- E2*CHIKV protein boost developed high levels of neutralizing antibodies, as well as specific IFN-γ producing cells and polyfunctional CD4+ and CD8+ T cells. We also identified 14 potential epitopes along the E2CHIKV protein. Furthermore, immunization with recombinant E2*CHIKV combined with the adjuvant AS03 presented the highest humoral response with neutralizing capacity. Finally, we show that the heterologous prime-boost strategy with the non-targeted pVAX-E2 DNA vaccine as the prime followed by E2* protein + AS03 boost is a promising combination to elicit a broad humoral and cellular immune response. Together, our data highlights the importance of E2CHIKV for the development of a CHIKV vaccine.- Published
- 2023
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9. Robust specific RBD responses and neutralizing antibodies after ChAdOx1 nCoV-19 and CoronaVac vaccination in SARS-CoV-2- seropositive individuals.
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Fernandes ER, Taminato M, de Souza Apostolico J, Gabrielonni MC, Lunardelli VAS, Maricato JT, Andersen ML, Tufik S, and Rosa DS
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Background: The pandemic unleashed by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected more than 500 million people worldwide and caused more than 6 million deaths. Cellular and humoral immunity induced by infection or immunization are key factors in controlling the viral burden and avoiding the recurrence of coronavirus disease. The duration and effectiveness of immunity after infection is relevant to pandemic policy interventions, including the timing of vaccine boosters., Objectives: We sought to evaluate longitudinal binding and functional antibodies against SARS-CoV-2 receptor-binding domain in police officers and health care workers with a history of coronavirus disease 2019 and compare with SARS-CoV-2-naive individuals after vaccination with adenovirus-based ChAdOx1 nCoV-19 (AstraZeneca-Fiocruz) or the inactivated CoronaVac vaccine (Sinovac-Butantan Institute)., Methods: A total of 208 participants were vaccinated. Of these, 126 (60.57%) received the ChAdOx1 nCoV-19 vaccine and 82 (39.42%) received the CoronaVac vaccine. Prevaccination and postvaccination blood was collected, and the amount of anti-SARS-CoV-2 IgG and the neutralizing ability of the antibodies to block the interaction between angiotensin-converting enzyme 2 and receptor-binding domain were determined., Results: Subjects with preexisting SARS-CoV-2 immunity and who received a single dose of ChAdOx1 nCoV-19 or CoronaVac have similar or superior antibody levels when compared with levels in seronegative individuals even after 2 doses of the vaccine. Neutralizing antibody titers of seropositive individuals were higher with a single dose of either ChAdOx1 nCoV-19 or CoronaVac compared with those of seronegative individuals. After 2 doses, both groups reached a plateau response., Conclusions: Our data reinforce the importance of vaccine boosters to increase specific binding and neutralizing SARS-CoV-2 antibodies., (© 2023 The Author(s).)
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- 2023
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10. Rabies virus isolated from insectivorous bats induces different inflammatory responses in experimental model.
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Motta GH, Guimarães LP, Fernandes ER, Guedes F, de Sá LRM, Dos Ramos Silva S, Ribeiro OG, and Katz ISS
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- Mice, Animals, Models, Theoretical, Rabies virus, Chiroptera, Rabies
- Abstract
Rabies virus (RABV) is a neurotropic virus that causes fatal neuroinflammation in mammals. The insectivorous bat RABV strains are less pathogenic for mice than strains associated with other reservoirs. We characterized the tissue inflammatory response in the CNS of RABV isolated from insectivorous bats. Eptesicus furinalis (EPBRV)-infected mice had a robust inflammatory response and a greater amount of IL-1β, IL-6 and TNF-α, while Myotis nigricans (MNBRV)-infected mice showed a higher expression of IL-17 and greater activation of IFN-β. New approaches to understand the inflammatory response to different mechanisms of action may provide insights for the development of novel therapies for rabies., Competing Interests: Declaration of Competing Interest All authors declare that they have no competing interests., (Copyright © 2022 Elsevier B.V. All rights reserved.)
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- 2022
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11. Oxygen saturation as a predictor of inflammation in obstructive sleep apnea.
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Fernandes ER, Pires GN, Andersen ML, Tufik S, and Rosa DS
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- Humans, Adult, Cross-Sectional Studies, Inflammation diagnosis, Inflammation complications, Lymphocytes, Oxygen Saturation, Sleep Apnea, Obstructive
- Abstract
Purpose: Obstructive sleep apnea (OSA) is a chronic sleep disorder, and its prevalence is increasing worldwide. This disorder has been consistently associated with several comorbidities. Although it is clear that obstructive sleep apnea severity is associated with inflammation, the trigger for this phenomenon continues to puzzle scientists. Here, we investigated the relationship between obstructive sleep apnea severity and immune parameters., Methods: In this cross-sectional epidemiological research, we analyzed the immune profile of 461 adults according to OSA severity (mild, moderate, and severe) and oxygen saturation., Results: The hallmark of OSA severity - the apnea-hypopnea index (AHI) - weakly correlated with an inflammatory profile. However, individuals who experienced lower oxygen saturation were more likely to exhibit higher total leukocyte and neutrophil counts, a higher neutrophil-lymphocyte ratio (NLR), and an increased concentration of C-reactive protein., Conclusion: Our findings indicated that oxygen saturation is a predictor of inflammation during OSA and should be considered crucial in disease diagnostic and treatment strategies., (© 2021. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)
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- 2022
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12. Time-dependent contraction of the SARS-CoV-2-specific T-cell responses in convalescent individuals.
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Fernandes ER, de Souza Apostolico J, Jacintho LC, Carnevale Marin ML, Vieira da Silva Júnior RC, Rodrigues H, Santos KS, Coelho V, Boscardin SB, Kalil J, Cunha-Neto E, and Rosa DS
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Background: Adaptive immunity in severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection is decisive for disease control. Delayed activation of T cells is associated with a worse outcome in coronavirus disease 2019 (COVID-19). Although convalescent individuals exhibit solid T-cell immunity, to date, long-term immunity to SARS-CoV-2 is still under investigation., Objectives: We aimed to characterize the specific T-cell response on the basis of the in vitro recall of IFN-γ-producing cells to in silico -predicted peptides in samples from SARS-CoV-2 convalescent individuals., Methods: The sequence of the SARS-CoV-2 genome was screened, leading to the identification of specific and promiscuous peptides predicted to be recognized by CD4
+ and CD8+ T cells. Next, we performed an in vitro recall of specific T cells from PBMC samples from the participants. The results were analyzed according to clinical features of the cohort and HLA diversity., Results: Our results indicated heterogeneous T-cell responsiveness among the participants. Compared with patients who exhibited mild symptoms, hospitalized patients had a significantly higher magnitude of response. In addition, male and older patients showed a lower number of IFN-γ-producing cells. Analysis of samples collected after 180 days revealed a reduction in the number of specific circulating IFN-γ-producing T cells, suggesting decreased immunity against viral peptides., Conclusion: Our data are evidence that in silico -predicted peptides are highly recognized by T cells from convalescent individuals, suggesting a possible application for vaccine design. However, the number of specific T cells decreases 180 days after infection, which might be associated with reduced protection against reinfection over time., (© 2022 The Authors.)- Published
- 2022
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13. Recurrence of COVID-19 associated with reduced T-cell responses in a monozygotic twin pair.
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de Castro MV, Santos KS, Apostolico JS, Fernandes ER, Almeida RR, Levin G, Magawa JY, Nunes JPS, Bruni M, Yamamoto MM, Lima AC, Silva MVR, Matos LRB, Coria VR, Castelli EC, Scliar MO, Kuramoto A, Bruno FR, Jacintho LC, Nunes K, Wang JYT, Coelho VP, Neto MM, Maciel RMB, Naslavsky MS, Passos-Bueno MR, Boscardin SB, Rosa DS, Kalil J, Zatz M, and Cunha-Neto E
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- Adolescent, Adult, Female, Humans, Male, Recurrence, COVID-19 immunology, Epitopes, T-Lymphocyte immunology, Immunity, Cellular, SARS-CoV-2 immunology, T-Lymphocytes immunology, Twins, Monozygotic
- Abstract
Recurrence of COVID-19 in recovered patients has been increasingly reported. However, the immune mechanisms behind the recurrence have not been thoroughly investigated. The presence of neutralizing antibodies (nAbs) in recurrence/reinfection cases suggests that other types of immune response are involved in protection against recurrence. Here, we investigated the innate type I/III interferon (IFN) response, binding and nAb assays and T-cell responses to severe acute respiratory distress syndrome coronavirus 2 (SARS-CoV-2) with IFN gamma (IFN γ ) enzyme-linked spot assay (ELISPOT) in three pairs of young adult monozygotic (MZ) twins with previous confirmed COVID-19, one of them presenting a severe recurrence four months after the initial infection. Twin studies have been of paramount importance to comprehend the immunogenetics of infectious diseases. Each MZ twin pair was previously exposed to SARS-CoV-2, as seen by clinical reports. The six individuals presented similar overall recovered immune responses except for the recurrence case, who presented a drastically reduced number of recognized SARS-CoV-2 T-cell epitopes on ELISPOT as compared to her twin sister and the other twin pairs. Our results suggest that the lack of a broad T-cell response to initial infection may have led to recurrence, emphasizing that an effective SARS-CoV-2-specific T-cell immune response is key for complete viral control and avoidance of clinical recurrence of COVID-19.
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- 2022
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14. A comparative review of serological assays for the detection of rabies virus-specific antibodies.
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Ciconello FN, Katz ISS, Fernandes ER, Guedes F, and Silva SR
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- Animals, Antibodies, Neutralizing, Antibodies, Viral, Humans, Neutralization Tests, Rabies diagnosis, Rabies prevention & control, Rabies Vaccines, Rabies virus
- Abstract
Rabies is a major public health problem with a fatality rate close to 100%, caused by a virus of the Lyssavirus genus, of which rabies virus (RABV) is the prototype. Nonetheless, the complete prevention can be achieved by the induction of neutralizing antibodies by pre- or post-exposure prophylaxis. According to the world health organization (WHO) and World Organization for animal health (OIE), serum titers of rabies virus neutralizing antibodies (RVNA) that are higher or equal to 0.5 international units (IU)/ml indicate adequate immune response after vaccination against rabies. Currently, RFFIT and FAVN are the gold standard tests recommended by both WHO and OIE for detecting and quantitating RVNA in biological samples from individuals or animals previously vaccinated and/or subjects suspected of having been infected by RABV. Although the tests RFFIT and FAVN are efficient, they are time-consuming, labor-intensive manual tests and not cost-effective for routine use. Following the previously mentioned, approaches with alternative methods have been developed to detect RVNA or rabies-specific antibodies in human or animal serum, but with variable success. This work summarizes the advances in the serological assays for the detection of neutralizing antibodies or rabies antibodies and assesses the individual immune status after vaccination against rabies, as well as the mechanisms of RABV neutralization mediated by antibodies. Therefore, the main alternative methods for the determination of RABV or rabies-specific antibodies are exposed, with promising results, besides being easy to execute, of low cost, and representing a possibility of being applied, according to the proposal of each test to the network of Rabies Surveillance Laboratories., (Copyright © 2021. Published by Elsevier B.V.)
- Published
- 2022
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15. Rabies Virus Exposure in Wild Lowland Tapirs (Tapirus terrestris) from Three Brazilian Biomes.
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Fernandes-Santos RC, Fernandes ER, Luiz FG, Chaves LB, Ramos Silva SD, and Medici EP
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- Animals, Antibodies, Viral blood, Brazil epidemiology, Rabies epidemiology, Rabies virology, Seroepidemiologic Studies, Ecosystem, Perissodactyla blood, Rabies veterinary, Rabies virus
- Abstract
We evaluated the presence of antibodies for rabies virus in 177 serum samples from 125 wild lowland tapirs (Tapirus terrestris) from three different Brazilian biomes. The rapid fluorescent focus inhibition test was performed. No antibody titers suggesting the circulation of the rabies virus in tapir habitat were detected., (© Wildlife Disease Association 2021.)
- Published
- 2021
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16. HIV-Infected Naïve Patients Exhibit Endothelial Dysfunction Concomitant with Decreased Natural Antibodies Against Defined Apolipoprotein B Autoantigens.
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Fonseca HAR, Gidlund M, Sant'Anna VR, Fernandes ER, Fonseca FAH, and Izar MC
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- Apolipoproteins B, Humans, Immunoglobulin G, Immunoglobulin M, Lipoproteins, LDL, Autoantigens, HIV Infections
- Abstract
Backgorund: Traditional and HIV-defined risk factors may be associated with an increase in cardiovascular events. Recent studies have suggested that the humoral immune response to modified LDL may be associated with the process of atherosclerosis., Objectives: To evaluate the presence of anti-oxLDL and apolipoprotein B-derived peptides in the blood, and their association with the endothelial function in HIV-infection., Methods: This study consecutively included subjects matched for age, gender, and demographic data in two groups: (1) HIV-infected and naïve for antiviral therapy and (2) uninfected individuals. Subclinical atherosclerosis was assessed by intimal-media thickness, using ultrasonography of the carotid arteries. Endothelial function was determined by flow-mediated dilatation (FMD) of the brachial artery by ultrasonography. Autoantibodies (IgM, IgG) anti-oxidized low-density lipoprotein (oxLDL), anti-apolipoprotein B-peptide fragments (ApoB-D and 0033G-Cys peptides), and cytokine levels were evaluated by ELISA., Results: This study's results showed no difference in subclinical atherosclerosis between groups; however, HIV-infected subjects showed a lower FMD, when compared to non-infected subjects. Therefore, HIV-infected subjects showed higher levels of inflammatory cytokines, titers of IgG anti-oxLDL, and IgG anti-ApoB-D. In contrast, titers of IgM anti-ApoB-D were lower in HIV-infected individuals and associated with reduced endothelial functions., Conclusions: This study's results show that HIV infection, in naïve subjects, is associated with endothelial dysfunction and a decline of natural antibodies to apo-B antigens.
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- 2021
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17. Prime-boost with Chikungunya virus E2 envelope protein combined with Poly (I:C) induces specific humoral and cellular immune responses.
- Author
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Amaral MP, Coirada FC, de Souza Apostolico J, Tomita N, Fernandes ER, Santos Souza HF, Chura-Chambi RM, Morganti L, Boscardin SB, and Rosa DS
- Abstract
Chikungunya virus (CHIKV) is an arbovirus transmitted to humans mainly by the bite of infected Aedes aegypti and Aedes albopictus mosquitoes. CHIKV illness is characterized by fever and long-lasting arthritic symptoms, and in some cases it is a deadly disease. The CHIKV envelope E2 (E2
CHIKV ) glycoprotein is crucial for virus attachment to the cell. Furthermore, E2CHIKV is the immunodominant protein and the main target of neutralizing antibodies. To date, there is no available prophylactic vaccine or specific treatment against CHIKV infection. Here, we designed and produced a DNA vaccine and a recombinant protein containing a consensus sequence of E2CHIKV . C57BL/6 mice immunized twice with the E2CHIKV recombinant protein in the presence of the adjuvant Poly (I:C) induced the highest E2CHIKV -specific humoral and cellular immune responses, while the immunization with the homologous DNA vaccine pVAX-E2CHIKV was able to induce specific IFN-γ producing cells. The heterologous prime-boost strategy was also able to induce specific cellular and humoral immune responses that were, in general, lower than the responses induced by the homologous E2CHIKV recombinant protein immunization. Furthermore, recombinant E2CHIKV induced the highest titers of neutralizing antibodies. Collectively, we believe this is the first report to analyze E2CHIKV -specific humoral and cellular immune responses after immunization with E2CHIKV recombinant protein and DNA pVAX-E2CHIKV vaccine platforms., Competing Interests: The authors declare that they have no conflicts of interest., (© 2021 The Author(s).)- Published
- 2021
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18. Zika virus-an update on the current efforts for vaccine development.
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Lunardelli VAS, Apostolico JS, Fernandes ER, and Santoro Rosa D
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- Adult, Female, Humans, Pregnancy, Guillain-Barre Syndrome epidemiology, Viral Vaccines, Zika Virus, Zika Virus Infection prevention & control
- Abstract
In 2015, the world witnessed the resurgence and global spread of Zika virus (ZIKV). This arbovirus infection is associated with Guillain-Barré syndrome in adults and with devastating congenital malformations during pregnancy. Despite scientific efforts, the development of a vaccine capable of inducing long-term protection has been challenging. Without a safe and efficacious licensed vaccine, control of virus transmission is based on vector control, but this strategy has been shown to be inefficient. An effective and protective vaccine relies on several requirements, which include: (i) induction of specific immune response against immunodominant antigens; (ii) selection of adjuvant-antigen formulation; and (iii) assessment of safety, effectiveness, and long-term protection. In this commentary, we provide a brief overview about the current efforts for the development of an efficacious ZIKV vaccine, covering the most important preclinical trials up to the formulations that are now being evaluated in clinical trials.
- Published
- 2021
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19. Glycosylation is required for the neutralizing activity of human IgG1 antibodies against human rabies induced by pre-exposure prophylaxis.
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Koike G, Katz ISS, Fernandes ER, Guedes F, and Silva SR
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- Glycosylation, Humans, Lectins immunology, Rabies virus immunology, Retrospective Studies, Antibodies, Neutralizing blood, Antibodies, Viral blood, Immunoglobulin G blood, Pre-Exposure Prophylaxis, Rabies prevention & control, Rabies Vaccines administration & dosage
- Abstract
Rabies lyssavirus (RABV) neutralizing IgG antibodies confer protection after rabies vaccination, although how the RABV-specific antibodies neutralize the virus is still unknown. As changes in the antibody's carbohydrate chain can interfere with its effector functions, we compared the glycosylation patterns of both neutralizing and non-neutralizing IgG1 induced by pre-exposure prophylaxis to human rabies and analyzed their influence on in vitro antibody neutralizing activities. Specific IgG1 were purified from human serum using affinity chromatography. Purity and avidity were analyzed by SDS-PAGE and indirect ELISA using NH
4 SCN respectively. The N-linked oligosaccharide chain of the purified IgG antibody was evaluated using a lectin-based ELISA assay with a panel of seven lectins. The activity of purified IgG1 and neutralizing IgG1 deglycosylated by PNGase F enzyme were analyzed using the rapid fluorescent focus inhibition test. The purified IgG1 showed an electrophoretic pattern compatible with human IgG. All of the antibodies recognized RABV, although neutralizing IgG1 had a higher avidity (RAI = 80%) than non-neutralizing IgG1 (RAI = 30%). The neutralizing IgG1 also showed higher binding to WFA, ECA, WGA, and ConA lectins, indicating possible different N-acetylgalactosamine, galactose, N-acetylglucosamine, and mannose contents. Non-neutralizing IgG1, on the other hand, showed strong binding at UEA-1 and SNA, which bind to fucose and sialic acid residues respectively. Different glycosylation profiles were also observed in Fab and Fc fragments from neutralizing and non-neutralizing IgG1, although the deglycosylated IgG1 lost its neutralizing activity. Our results suggest that antibody glycosylation is important for neutralizing RABV in vitro, since neutralizing IgG1 has a different glycosylation profile than non-neutralizing IgG1. Further research will be needed to better evaluate the differential glycosylation patterns between IgG1 antibodies following vaccination., (Copyright © 2021 Elsevier GmbH. All rights reserved.)- Published
- 2021
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20. Invariant Natural Killer T cells resilience to paradoxical sleep deprivation-associated stress.
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Sousa MEP, Gonzatti MB, Fernandes ER, Freire BM, Guereschi MG, Basso AS, Andersen ML, Rosa DS, and Keller AC
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- Animals, Cytokines, Killer Cells, Natural, Mice, Mice, Inbred C57BL, Sleep, REM, Spleen, Natural Killer T-Cells
- Abstract
Although several studies demonstrate that stressful situations, such as sleep disturbances, negatively impact the innate and adaptive arms of the immune system, their influence on invariant Natural Killer T (iNKT) cells remains unclear. iNKT cells are CD1d-restricted innate T cells that recognize glycolipid antigens and rapidly produce polarizing cytokines being key players in several immune responses, and a potential target for immunotherapy. iNKT cells differ in several aspects from conventional T lymphocytes, including a unique dependence on CD1d-expressing double-positive (DP) thymocytes for intrathymic maturation. As a consequence of stress, DP thymocytes undergo glucocorticoid-induced apoptosis, which might compromise iNKT developmental pathway. Therefore, we used a paradoxical sleep deprivation (SD) model to determine the impact of sleep disturbance on iNKT cell biology. After 72 h of SD, C57Bl/6 mice exhibited a significant increase in systemic glucocorticoid levels and thymus atrophy. Despite marked decrease in the number of DP thymocytes, the ratio CD1d
+ /CD1d- was higher in SD mice, and the number of thymic iNKT cells remained unaltered, suggesting that SD did not compromise the iNKT developmental pathway. In contrast, SD reduced hepatic IFN-γ, but not, IL-4-producing iNKT cells, without further effect in the spleen. Despite this fact, SD did not affect stimulation of IFN-γ production by iNKT cells, or cytokine release, in response to α-galactosylceramide, a specific antigen. Furthermore, although SD impaired splenic NK cells activity against tumor cells, it did not affect iNKT cell-specific cytotoxicity. Thus, our study shows that SD-induced stress did not impair the iNKT cells' responses to a cognate antigen., (Copyright © 2020 Elsevier Inc. All rights reserved.)- Published
- 2020
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21. Development of biotinylated polyclonal anti-ribonucleoprotein IgG for detection of rabies virus antigen by direct rapid immunohistochemical test.
- Author
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Castro BS, Guedes F, Fernandes ER, Koike G, Katz ISS, Chaves LB, and Silva SR
- Subjects
- Animals, Antibodies, Monoclonal metabolism, Antibodies, Viral immunology, Antibodies, Viral metabolism, Biotinylation, Brain immunology, Brain virology, Cats, Cattle, Chiroptera, Dogs, Fluorescent Antibody Technique, Direct methods, Horses, Immunoglobulin G metabolism, Immunohistochemistry methods, Primates, Rabies diagnosis, Rabies virology, Sensitivity and Specificity, Species Specificity, Swine, Antibodies, Monoclonal immunology, Antigens, Viral immunology, Immunoglobulin G immunology, Rabies immunology, Rabies virus immunology, Ribonucleoproteins immunology
- Abstract
The direct rapid immunohistochemical test (dRIT) has been recommended for laboratorial diagnosis of rabies, especially in developing countries. The absence of commercial primary antibodies, however, still represents a major limitation to its wider use in testing. We describe here the development of a biotinylated polyclonal antibody against Rabies lyssavirus (RABV) ribonucleoprotein (RNP) and its use as a primary reagent in dRIT. Anti-RNP polyclonal horse IgG was purified by ionic exchange chromatography followed by immunoaffinity column chromatography, and its affinity, diagnostic sensitivity, and specificity were evaluated. CNS samples (120) of suspected rabies cases in different animal species were tested by dRIT, with the positive (n = 14) and negative (n = 106) results confirmed by direct fluorescence antibody testing (dFAT). Comparing the results of dRIT and dFAT, we found that the biotinylated anti-RNP IgG delivered 100% diagnostic specificity and sensibility for rabies diagnosis. Our findings show that the biotinylated anti-RNP polyclonal IgG can be produced with the quality required for application in dRIT. This work represents an important step in efforts to diagnose rabies in developing countries., (Copyright © 2020 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2020
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22. Sleep Disturbance during Infection Compromises Tfh Differentiation and Impacts Host Immunity.
- Author
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Fernandes ER, Barbosa ML, Amaral MP, de Souza Apostolico J, Sulczewski FB, Tufik S, Andersen ML, Boscardin SB, Keller AC, and Rosa DS
- Abstract
Although the influence of sleep quality on the immune system is well documented, the mechanisms behind its impact on natural host immunity remain unclear. Meanwhile, it has been suggested that neuroimmune interactions play an important role in this phenomenon. To evaluate the impact of stress-induced sleep disturbance on host immunity, we used a murine model of rapid eye movement sleep deprivation (RSD) integrated with a model of malaria blood-stage infection. We demonstrate that sleep disturbance compromises the differentiation of T follicular helper cells, increasing host susceptibility to the parasite. Chemical inhibition of glucocorticoid (Glcs) synthesis showed that abnormal Glcs production compromised the transcription of Tfh-associated genes resulting in impaired germinal center formation and humoral immune response. Our data demonstrate that RSD-induced abnormal activation of the hypothalamic-pituitary-adrenal axis drives host susceptibility to infection. Understanding the impact of sleep quality in natural resistance to infection may provide insights for disease management., Competing Interests: The authors declare no competing interests., (© 2020.)
- Published
- 2020
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23. Detection of rabies virus antigen by the indirect rapid immunohistochemistry test in equines and comparisons with other diagnostic techniques.
- Author
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Torquato RBC, Iamamoto K, Fernandes ER, Achkar S, Silva SR, Katz ISS, and Guedes F
- Subjects
- Animals, Central Nervous System virology, Horse Diseases virology, Horses, Immunohistochemistry methods, Neurons virology, Rabies diagnosis, Rabies virus immunology, Antigens, Viral isolation & purification, Horse Diseases diagnosis, Immunohistochemistry veterinary, Rabies veterinary, Rabies virus isolation & purification
- Abstract
Laboratory diagnosis of rabies in equines is essential for distinguishing the disease from other sources of encephalitis. Diagnosis by conventional techniques such as a direct fluorescent antibody test (dFAT) or viral isolation in mice or cell culture can be difficult, and the application of molecular biological methods may be necessary. We performed an indirect rapid immunohistochemistry test (iRIT) for the detection of the rabies virus (RABV) antigen in the central nervous system (CNS) of equines and compared the results with those of other diagnostic techniques. We reviewed result records from the Rabies Diagnosis Laboratory at Instituto Pasteur, São Paulo, Brazil, of 174 samples of equine CNS from July 2014 to June 2016, which were investigated by dFAT, rabies tissue culture infection test (RTCIT), mouse inoculation test (MIT) and reverse transcription-polymerase chain reaction (RT-PCR) followed by genetic sequencing. These samples, 29 presented divergent results among techniques and were selected for the performed in the iRIT. The detected positivity rate was 4/29 (14%) by dFAT, 5/28 (18%) by RTCIT, 10/29 (35%) by MIT and 26/27 (96%) by RT-PCR. We analysed 29 samples through imprints of the cortex, hippocampus, cerebellum and brainstem in slides fixed in 10% buffered formaldehyde. Eighteen samples were identified as positive (62%) by iRIT assay, representing a greater number of positive cases than that detected by dFAT, MIT and RTCIT but not by RT-PCR. Among the brain regions, the brainstem presented the highest positivity (78%), followed by the hippocampus (69%), cerebellum (67%) and cortex (67%). Our results provide evidence that iRIT can contribute to a rapid diagnosis of rabies in equines and that complementary tests should be used to improve diagnostic accuracy in this species., (© 2020 Blackwell Verlag GmbH.)
- Published
- 2020
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24. Characterization of the Th17 profile immune response in cases of human rabies transmitted by dogs and its interference in the disease pathogenesis.
- Author
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Santos LB, Guedes F, Achkar SM, Duarte MIS, Katz ISS, Silva SR, and Fernandes ER
- Subjects
- Adolescent, Adult, Animals, Brain metabolism, Cytokines metabolism, Dogs, Female, Humans, Male, Middle Aged, Rabies metabolism, Th17 Cells metabolism, Young Adult, Brain immunology, Cytokines immunology, Immunity, Cellular immunology, Rabies immunology, Rabies transmission, Th17 Cells immunology
- Abstract
The Th17 profile immune response is influenced by the presence of cytokines such as IL-1, IL-6, TGF-β, IL-17, and IL-23. We sought to characterize the Th17 profile in CNS samples from human rabies cases transmitted by dogs and examine its possible influence on disease pathogenesis. We observed a high expression of TGF-β, followed by IL-23, IL-17 and IL-6, and a low expression of IL-1β and IFN-γ. Those results suggest the participation of Th17 in rabies virus neuroinfection transmitted by dogs. IL-23 probably plays a role in maintaining the Th17 profile, but it can also interfere with the establishment of the Th1 profile and viral clearance., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
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25. Performance evaluation of the polyclonal anti-rabies virus ribonucleoprotein IgG antibodies produced in-house for use in direct fluorescent antibody test.
- Author
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da Silva GH, Santos da Silva JH, Iamamoto K, de Arruda TS, Katz ISS, Fernandes ER, Guedes F, Rodrigues da Silva AC, and Silva SR
- Subjects
- Animals, Antibodies, Viral chemistry, Antigens, Viral analysis, Antigens, Viral immunology, Brazil, Fluorescein-5-isothiocyanate chemistry, Fluorescent Antibody Technique, Direct, Immunoglobulin G chemistry, Immunoglobulin G immunology, Rabies virus immunology, Reproducibility of Results, Sensitivity and Specificity, Antibodies, Viral immunology, Rabies diagnosis, Rabies virus isolation & purification, Ribonucleoproteins immunology
- Abstract
Fluorescein isothiocyanate (FITC) labelled anti-rabies virus ribonucleoprotein (RNP) antibodies can be used as immunoreagents in direct fluorescent antibody testing (dFAT) for rabies diagnoses. While in-house products are occasionally used by laboratories, most conjugates are commercial reagents. Commercial anti-RNP antibodies are only available for research purposes in Brazil, however, which contributes to the increasing use of in-house produced antibodies. Considering that conjugate quality may influence the results obtained during rabies diagnosis, we sought to analyze the performance requirements of in-house produced polyclonal anti-RNP IgG-FITC for application in dFAT. To that end, their reproducibility, diagnostic sensitivity, and specificity were evaluated. The titer of polyclonal anti-RNP IgG-FITC was initially determined and evaluated by dFAT, using central nervous system (CNS) samples of different animal species (dogs, cats, bovines, equines, bats, and non-human primates). As our main result, the polyclonal anti-RNP IgG-FITC reached a titer of 1:30/1:40 in dFAT, with 100% of diagnostic sensitivity and specificity. In terms of reproducibility, the antibodies, regardless the production batch, presented the same performances. In conclusion, the in-house produced polyclonal anti-RNP IgG-FITC proved suitable for rabies virus antigen detection by dFAT., Competing Interests: Declaration of Competing Interest The authors declare that they have no financial or personal relationships that may have inappropriately influenced them in writing this article., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
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26. Evaluation of polyclonal anti-RNP IgG antibody for rabies diagnosis by indirect rapid immunohistochemistry test.
- Author
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Bartaquini RT, Torquato RB, Fernandes ER, Guedes F, de Castro BS, Katz ISS, Scheffer KC, da Silva ACR, Pimenta DC, and Silva SR
- Subjects
- Animals, Fluorescent Antibody Technique, Direct, Humans, Antibodies, Viral immunology, Immunoglobulin G immunology, Immunohistochemistry methods, Rabies diagnosis, Rabies virus immunology, Ribonucleoproteins immunology
- Abstract
Rabies still represents a major public health threat and estimated to cause 60,000 human deaths annually, particularly in developing countries. Thus, adequate surveillance based on rapid and reliable rabies diagnosis for both humans and animals is essential. The WHO and OIE recommended gold standard diagnostic technique for rabies is the direct immunofluorescence assay (dFAT). However, dFAT is expensive and requires a high level of expertise. As an alternative, the rapid immunohistochemistry technique is a promise to be a simple and cost effective diagnostic tool for rabies, and can be performed on field conditions prevalent in developing countries. However, no validated commercial conjugate antibody for rabies is available to meet the laboratory demand. Here, we evaluated the polyclonal anti-rabies virus ribonucleoprotein (RNP) IgG antibody for Rabies lyssavirus (RABV) detection by indirect rapid immunohistochemistry test (iRIT). We tested polyclonal anti-RNP IgG antibody against a batch of 100 brain specimens representing a wide phylogenetic origin in the State of São Paulo, Brazil. The purified IgG obtained 100% of diagnostic specificity and sensibility for RABV antigen detection in iRIT compared with the gold standard dFAT. In conclusion, our results demonstrate that the polyclonal anti-RNP IgG antibody may be used as a diagnostic reagent for rabies using iRIT, with the expectation of increase in availability and cost reduction of the epidemiological surveillance for developing countries., Competing Interests: Declaration of Competing Interest None, (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2020
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27. Homologous prime-boost with Zika virus envelope protein and poly (I:C) induces robust specific humoral and cellular immune responses.
- Author
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Amaral MP, Apostolico JS, Tomita N, Coirada FC, Lunardelli VAS, Fernandes ER, Souza HFS, Astray RM, Boscardin SB, and Rosa DS
- Subjects
- Animals, Immunity, Cellular, Immunization, Secondary, Mice, Mice, Inbred BALB C, Viral Envelope, Vaccines, DNA, Viral Envelope Proteins, Zika Virus immunology, Zika Virus Infection prevention & control
- Abstract
The recent outbreaks of Zika virus (ZIKV) infection and the potential association with Guillain-Barré syndrome in adults and with congenital abnormalities have highlighted the urgency for an effective vaccine. The ZIKV Envelope glycoprotein (E
ZIKV ) is the most abundant protein on the virus surface, and has been evaluated together with the pre-membrane protein (prM) of the viral coat as a vaccine candidate in clinical trials. In this study, we performed a head-to-head comparison of the immune response induced by different EZIKV -based vaccine candidates in mice. We compared different platforms (DNA, recombinant protein), adjuvants (poly (I:C), CpG ODN 1826) and immunization strategies (homologous, heterologous). The hierarchy of adjuvant potency showed that poly (I:C) was a superior adjuvant than CpG ODN. While poly (I:C) assisted immunization reached a plateau in antibody titers after two doses, the CpG ODN group required an extra immunization dose. Besides, the administration of poly (I:C) induced higher EZIKV -specific cellular immune responses than CpG ODN. We also show that immunization with homologous prime-boost EZIKV protein + poly (I:C) regimen induced a more robust humoral response than homologous DNA (pVAX-EZIKV ) or heterologous regimens (DNA/protein or protein/DNA). A detailed analysis of cellular immune responses revealed that homologous (EZIKV + poly (I:C)) and heterologous (pVAX-EZIKV /EZIKV + poly (I:C)) prime-boost regimens induced the highest magnitude of IFN-γ secreting cells and cytokine-producing CD4+ T cells. Overall, our data demonstrate that homologous EZIKV + poly (I:C) prime-boost immunization is sufficient to induce more robust specific-EZIKV humoral and cellular immune responses than the other strategies that contemplate homologous DNA (pVAX-EZIKV ) or heterologous (pVAX-EZIKV /EZIKV + poly (I:C), and vice-versa) immunizations., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)- Published
- 2020
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28. Nyctinomops laticaudatus bat-associated Rabies virus causes disease with a shorter clinical period and has lower pathogenic potential than strains isolated from wild canids.
- Author
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Fuoco NL, Fernandes ER, Guedes F, Dos Ramos Silva S, Guimarães LP, Silva NU, Ribeiro OG, and Katz ISS
- Subjects
- Animals, Cell Line, Central Nervous System pathology, Disease Models, Animal, Histocytochemistry, Mice, Neurons virology, Survival Analysis, Virulence, Virus Replication, Canidae virology, Chiroptera virology, Rabies pathology, Rabies virology, Rabies virus isolation & purification, Rabies virus pathogenicity
- Abstract
Rabies is a lethal viral disease that can affect a wide range of mammals. Currently, Rabies virus (RABV) in some European and American countries is maintained primarily in wild species. The regulation of viral replication is one of the critical mechanisms involved in RABV pathogenesis. However, the relationship between replication and the pathogenesis of RABV isolated from wild animals remains poorly understood. In the present study, we evaluated the pathogenicity of the street viruses Nyctinomops laticaudatus bat-associated RABV (NYBRV) and Cerdocyon thous canid-associated RABV (CECRV). Infection of mice with NYBRV led to 33% mortality with rapid disease evolution and marked histopathological changes in the CNS. In contrast, infection with CECRV led to 67% mortality and caused mild neuropathological lesions. The proportion of RABV antigen was significantly higher in the cytoplasm of neuronal cells of the cerebral cortex and in the meninges of mice infected with CECRV and NYBRV, respectively. Moreover, the replication rate of NYBRV was significantly higher (p < 0.001) than that of CECRV in neuroblastoma cells. However, CECRV replicated to a significantly higher titer in epithelial cells. Our results indicate that NYBRV infection results in rapid disease progression accompanied by frequent and intense histopathological alterations in the CNS in mice, and in a high replication rate in neuroblastoma cells. Although, CECRV is more pathogenic in mice, it caused milder histopathological changes in the CNS and replicated more efficiently in epithelial cells. Our data point to a correlation between clinical aspects of disease and the replication of RABV in different cell lines.
- Published
- 2019
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29. Purification of IgG against ribonucleoprotein by a homemade immunoaffinity chromatography column for rabies diagnosis.
- Author
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da Silva Santos JH, da Silva GH, Iamamoto K, Katz ISS, Guedes F, Fernandes ER, de Cassia Rodrigues da Silva A, and Dos Ramos Silva S
- Subjects
- Animals, Antibodies, Monoclonal immunology, Antibodies, Viral immunology, Antigens, Viral immunology, Cats, Cattle, Cell Line, Chiroptera, Dogs, Haplorhini, Horses, Rabies immunology, Rabies virology, Rabies virus physiology, Sensitivity and Specificity, Species Specificity, Antibodies, Antinuclear immunology, Chromatography, Affinity methods, Immunoglobulin G immunology, Rabies diagnosis, Rabies virus immunology, Ribonucleoproteins immunology
- Abstract
Polyclonal or monoclonal antibodies against rabies virus ribonucleoprotein (RNP) conjugated to fluorescein isothiocyanate (FITC) have been employed for Rabies virus (RABV) antigen detection by the direct fluorescent antibody test (DFA). To date, these biomolecules have been purified by traditional methods such as precipitation by ammonium sulfate or ion exchange chromatography followed by ammonium sulfate precipitation, which allows only for partial detection of the protein of interest. In this study, we aimed to purify anti-RNP polyclonal horse IgG antibodies by cation-exchange chromatography in combination with a homemade immunoaffinity chromatography on RNP immobilized (RNP-IAC). Furthermore, to evaluate the accuracy of the prepared anti-RNP IgG fluorescent antibody in diagnostic purposes, DFA was applied for RABV antigen detection in suspected brain samples of different animal species. The combination of these two techniques made it possible to obtain antibodies with high selectivity and purity. Compared with the performance of the traditional method, anti-RNP IgG antibodies purified by RNP-IAC can be obtained from a smaller volume of hyperimmune serum and with greater avidity. Furthermore, the results obtained by DFA analyses revealed that the prepared anti-RNP IgG fluorescent antibody achieved 100% diagnostic specificity and sensitivity for RABV antigen detection. Thus, two-technique chromatographic, including RNP-IAC technology could be appropriate methods for the purification of polyclonal anti-RNP IgG for the use as a diagnostic reagent for rabies., (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Published
- 2019
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30. Extrinsically magnetic poly(butylene succinate): An up-and-coming petroleum cleanup tool.
- Author
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Figueiredo AS, Icart LP, Marques FD, Fernandes ER, Ferreira LP, Oliveira GE, and Souza FG Jr
- Abstract
This work presents the synthesis and characterization of extrinsically magnetic poly(butylene succinate) (PBS). PBS is obtained from succinic acid (SA), which can be efficiently produced from renewable biomass by fermentation. Thus, the use of SA helps to remove CO
2 from the atmosphere, constituting a good way to accumulate carbon credits. The magnetic PBS here presented was prepared by fusion using different amounts of maghemite. Obtained materials were characterized using Fourier transform infrared spectroscopy (FTIR), Thermogravimetric analysis (TGA), Differential scanning calorimetry (DSC), X-ray diffraction (XRD), Small angle X-ray scattering and magnetic force tests. Besides, the oil removal capability (OR) of the samples was also studied. All the magnetic composites were able to remove petroleum from the water. Among them, the one filled with the highest amount of magnetic particles was able to remove 11 g of oil per gram of composite. Also, XRD and SAXS results showed that PBS is a long size oriented material, which allows it to work as a thermoset, avoiding its dissolution in organic contaminant medium. As PBS can also be considered as a platform, these are promising results for the oil spill cleanup applications., (Copyright © 2018 Elsevier B.V. All rights reserved.)- Published
- 2019
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31. Street rabies virus strains associated with insectivorous bats are less pathogenic than strains isolated from other reservoirs.
- Author
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Fuoco NL, Fernandes ER, Dos Ramos Silva S, Luiz FG, Ribeiro OG, and Santos Katz IS
- Subjects
- Animals, Disease Models, Animal, Mice, Rabies virus growth & development, Rabies virus isolation & purification, Survival Analysis, Time Factors, Virulence, Virus Internalization, Virus Release, Virus Replication, Chiroptera virology, Rabies pathology, Rabies virology, Rabies virus pathogenicity
- Abstract
Rabies is a fatal and viral zoonosis that causes acute, progressive encephalitis and remains an important concern in public health. In the last few years, there has been a change in the epidemiological profile of rabies after implementing canine rabies control in the Americas, which has led to a significant increase in both human and pet cases of rabies associated with insectivorous bats. Thus, it is important to understand the pathogenesis caused by Rabies virus (RABV) isolates from insectivorous bats. Viral growth kinetics, cell-to-cell spread and virus uptake in vitro were analyzed for RABV isolates from Eptesicus furiralis and Myotis nigricans. For pathogenesis evaluation, mice were inoculated with RABV isolates from Eptesicus furiralis and Myotis nigricans, and clinical signs were observed for 40 days. We observed that the insectivorous bat strains showed a higher replication rate, faster cell-to-cell spread and delayed virus uptake in N2a cells. Furthermore, after the first sign of a clinical infection, mice infected with Myotis nigricans and Eptesicus furiralis isolates succumbed rapidly (6 ± 9 days) compared with RABV strains associated with other reservoirs. Our results show that the insectivorous bat RABV strains are less pathogenic for mice than strains associated with other reservoirs. In addition, this study also indicates that the differences in the biological characteristics of the RABV strains are important to their pathogenicity. An enhanced understanding of rabies pathogenesis may be important for the development of novel therapies for humans and in the implementation of rabies control strategies., (Copyright © 2018 Elsevier B.V. All rights reserved.)
- Published
- 2018
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32. Sleep deprivation predisposes allergic mice to neutrophilic lung inflammation.
- Author
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Nunes JOF, Apostolico JS, Andrade DAG, Ruiz FS, Fernandes ER, Andersen ML, Keller AC, and Rosa DS
- Subjects
- Animals, Cytokines genetics, Cytokines immunology, Disease Susceptibility, Female, Humans, Hypersensitivity genetics, Hypersensitivity pathology, Inflammation genetics, Inflammation immunology, Inflammation pathology, Mice, Mice, Knockout, Pneumonia genetics, Pneumonia pathology, Sleep Deprivation genetics, Sleep Deprivation pathology, Th17 Cells pathology, Hypersensitivity immunology, Pneumonia immunology, Sleep Deprivation immunology, Th17 Cells immunology
- Abstract
Background: Although different studies associated sleep deprivation (SD) with systemic inflammatory changes, the effect of sleep duration on the pathology of allergic chronic diseases is poorly understood., Objective: We sought to evaluate the influence of SD on allergen-induced pulmonary inflammation., Methods: Ovalbumin (OVA)-sensitized C57BL/6 mice were exposed to a first set of intranasal OVA challenge under SD or healthy sleep (HS) conditions, followed by a second OVA challenge, 1 week apart. Some groups were subjected to corticosteroid treatment with dexamethasone., Results: OVA-sensitized mice with SD had more severe airway inflammation than the allergic group with HS. Analysis of lung parenchyma revealed that the inflammation in allergic mice with SD was marked by an influx of neutrophils (mainly) and eosinophils and secretion of IL-6, TNF-α, and IL-17 in contrast to the eosinophilic inflammation and IL-4 production observed in allergic mice with HS. The same cytokine profile was observed in ex vivo culture of cervical lymph node cells and splenocytes, indicating that in allergic mice SD favors immune responses toward a proinflammatory T
H 17 profile. This idea is supported by the fact that disruption of IL-17 signaling (IL-17 receptor A-/- ) prevented airway neutrophilia in allergic mice with SD. Furthermore, allergic mice with SD became refractory to corticosteroid treatment in contrast to the allergic group with HS., Conclusion: Collectively, our data show that sleep quality participates in the progression of allergen-induced eosinophilic lung inflammation to corticosteroid-refractory neutrophilic manifestation., (Copyright © 2017 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)- Published
- 2018
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33. Infection of neuroblastoma cells by rabies virus is modulated by the virus titer.
- Author
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Fuoco NL, Dos Ramos Silva S, Fernandes ER, Luiz FG, Ribeiro OG, and Katz ISS
- Subjects
- Animals, Brain virology, Cell Line, Tumor, Cells, Cultured, Mice, Neuroblastoma, Viral Load, Virus Internalization, Rabies virology, Rabies virus physiology, Virus Replication
- Abstract
Rabies is a lethal viral infection that can affect almost all mammals, including humans. To better understand the replication of Rabies lyssavirus, we investigated if the viral load in brains naturally infected with rabies influences viral internalization and viral growth kinetics in neuroblastoma cells, and if the viral load affects mortality in mice after intradermal infection. We noted that high initial viral loads in brains (group II) were unfavourable for increasing viral titers during serial passages in neuroblastoma cells when compared to low initial viral loads in brains (group I). In addition, group I strains showed higher viral growth and enhanced internalization efficiency in neuroblastoma cells than group II strains. However, we observed that the dominant virus subpopulation in group II promoted efficient viral infection in the central nervous system in the new host, providing a selective advantage to the virus. Our data indicate that rabies infection in animal models depends on not only the virus strain but also the amount of virus. This study may serve as a basis for understanding the biologic proprieties of Rabies lyssavirus strains with respect to the effects on viral replication and the impact on pathogenesis, improving virus yields for use in vaccine development., (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Published
- 2018
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34. Immunological aspects of rabies: a literature review.
- Author
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Katz ISS, Guedes F, Fernandes ER, and Dos Ramos Silva S
- Subjects
- Animals, Humans, Zoonoses, Antibodies, Neutralizing immunology, Antibodies, Viral immunology, Rabies immunology, Rabies virus immunology
- Abstract
Rabies is a lethal disease caused by the neurotropic virus rabies virus (RABV), and it remains an important public health problem globally. It is known that the host immune response is important for control of viral infection and promoting viral clearance. In this context, it is well documented that, in addition to RABV neutralizing antibody, interferons and cell-mediated immunity also have an important role in preventing the establishment of disease. On the other hand, RABV suppresses host immunity through different mechanisms, for example, direct inhibition of host gene expression, sequestration of pathogen-associated molecular patterns, or modification of cytokine signalling pathways, which hinder the protective host immune responses to RABV infection. Here, we review the immunological aspects of rabies, highlighting innate and adaptive immunity, as well as the host evasion immune mechanisms used by the virus. Finally, we briefly discuss how this knowledge can direct new research and be harnessed for future therapeutic strategies.
- Published
- 2017
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35. Use of medicines recommended for secondary prevention of acute coronary syndrome.
- Author
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Gaedke MÂ, Costa JS, Manenti ER, Henn RL, Paniz VM, Nunes MF, Motta MA, and Olinto MT
- Subjects
- Adult, Aged, Brazil, Cardiovascular Agents classification, Cohort Studies, Drug Therapy, Combination, Evidence-Based Medicine, Female, Humans, Male, Middle Aged, Practice Guidelines as Topic, Secondary Prevention, Acute Coronary Syndrome prevention & control, Cardiovascular Agents administration & dosage
- Abstract
Objective: To analyze if the demographic and socioeconomic variables, as well as percutaneous coronary intervention are associated with the use of medicines for secondary prevention of acute coronary syndrome., Methods: In this cohort study, we included 138 patients with acute coronary syndrome, aged 30 years or more and of both sexes. The data were collected at the time of hospital discharge, and after six and twelve months. The outcome of the study was the simultaneous use of medicines recommended for secondary prevention of acute coronary syndrome: platelet antiaggregant, beta-blockers, statins and angiotensin-converting-enzyme inhibitor or angiotensin receptor blocker. The independent variables were: sex, age, education in years of attending, monthly income in tertiles and percutaneous coronary intervention. We described the prevalence of use of each group of medicines with their 95% confidence intervals, as well as the simultaneous use of the four medicines, in all analyzed periods. In the crude analysis, we verified the outcome with the independent variables for each period through the Chi-square test. The adjusted analysis was carried out using Poisson Regression., Results: More than a third of patients (36.2%; 95%CI 28.2;44.3) had the four medicines prescribed at the same time, at the moment of discharge. We did not observe any differences in the prevalence of use in comparison with the two follow-up periods. The most prescribed class of medicines during discharge was platelet antiaggregant (91.3%). In the crude analysis, the demographic and socioeconomic variables were not associated to the outcome in any of the three periods., Conclusions: The prevalence of simultaneous use of medicines at discharge and in the follow-ups pointed to the under-utilization of this therapy in clinical practice. Intervention strategies are needed to improve the quality of care given to patients that extend beyond the hospital discharge, a critical point of transition in care.
- Published
- 2015
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36. Immunopathogenesis of dengue hemorrhagic fever: contribution to the study of human liver lesions.
- Author
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Pagliari C, Quaresma JA, Fernandes ER, Stegun FW, Brasil RA, de Andrade HF Jr, Barros V, Vasconcelos PF, and Duarte MI
- Subjects
- Apoptosis, Cytokines analysis, Hepatocytes immunology, Humans, Immunohistochemistry, Immunologic Factors analysis, Lymphocyte Activation, Lymphocyte Subsets immunology, Microscopy, Liver pathology, Severe Dengue immunology, Severe Dengue pathology
- Abstract
Dengue infection is an important tropical disease worldwide. The host immune response has been studied in order to better understand lesion mechanisms. It was performed an immunohistochemical study in 14 specimens of liver from patients with dengue hemorrhagic fever (DHF) to characterize cytokines and some factors present in liver lesions and their possible role in the pathogenesis of hepatic injury. Portal tract and hepatic acinus presented high expression of TLR2, TLR3, IL6, and granzyme B. Hepatic acinus also presented iNOS, IL18, and TGF-beta. Cells expressing IL12, IL13, JAk1, STAT1, and NF-κB were rarely visualized. Treg cells foxp3+ were absent. TLR2 and TLR3 seem to participate in cellular activation and cytokine production. Cytotoxic response seems to play a role. Although TGF-beta promotes the activation of Foxp3+ regulatory T cells, IL6 can significantly suppresses their generation. The expression of Treg cells is diminished probably as a result of the high frequency of these cytokines. Both cytokines play a role in the increased vascular permeability and edema observed in dengue liver specimens, with consequent plasma leakage and severity of the disease. It was observed a regular expression of IL-18 in hepatocytes and lymphocytes of the inflammatory infiltrate in portal tract, which reflects the acute inflammatory response that occurs in the liver and contributes to hepatic injury. At least in part, the increased number of cells expressing IL-18 could play a role of "up" regulation of FasL and correlate to the phenomenon of apoptosis, a mechanism of destruction of hepatocytes in DHF., (© 2013 Wiley Periodicals, Inc.)
- Published
- 2014
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37. Behavioral changes after cardiovascular events: a cohort study.
- Author
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Franken DL, Olinto MT, Paniz VM, Henn RL, Junqueira LD, da Silveira FG, Roman VR, Manenti ER, and Dias da Costa JS
- Subjects
- Adult, Aged, Cohort Studies, Female, Humans, Male, Cardiovascular Diseases prevention & control, Risk Reduction Behavior
- Published
- 2012
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38. Expression of TLR2 and TLR4 in lesions of patients with tegumentary American leishmaniasis.
- Author
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Tuon FF, Fernandes ER, Duarte MI, and Amato VS
- Subjects
- Adolescent, Adult, Cell Count, Dendritic Cells metabolism, Female, Humans, Immunohistochemistry, Interferon Type I analysis, Interferon Type I immunology, Interleukin-1 analysis, Interleukin-1 immunology, Interleukin-6 analysis, Interleukin-6 immunology, Killer Cells, Natural metabolism, Leishmaniasis, Cutaneous immunology, Macrophages metabolism, Male, Middle Aged, Polymerase Chain Reaction, Prospective Studies, Toll-Like Receptor 2 immunology, Toll-Like Receptor 4 immunology, Tumor Necrosis Factor-alpha analysis, Tumor Necrosis Factor-alpha immunology, Young Adult, Cytokines analysis, Leishmaniasis, Cutaneous metabolism, Toll-Like Receptor 2 metabolism, Toll-Like Receptor 4 metabolism
- Abstract
Objectives: The aim of this study was to describe the pattern of expression of Toll-like receptor 2 (TLR2) and Toll-like receptor 4 (TLR4) in skin biopsies of patients with American tegumentary leishmaniasis (ATL) caused by Leishmania braziliensis., Methods: This prospective study evaluated 12 patients with ATL caused by Leishmania braziliensis confirmed by polymerase chain reaction. Immunohistochemistry was performed to determine the expression of TLR2 and TLR4. The number of NK cells, dendritic cells and macrophages in the tissue were calculated. The cytokine expression was determined using the anti-TNF-α, anti-IFN-Γ, anti-IL-1 and anti-IL-6. Double immunostaining reactions were used to determine the cell expressing TLR2 and TLR4., Results: The numbers of cells expressing TLR2 and TLR4 were 145.48 ± 82.46 cell/mm² and 3.26 ± 4.11 cell/mm² respectively (p < 0.05). There was no correlation of TLR2 and TLR4 with the amount of cytokines and the number of NK cells, dendritic cells or macrophages. The double immunostaining revealed that TLR2 was expressed by macrophages., Conclusion: In human cutaneous leishmaniasis caused by Leishmania braziliensis, TLR2 is the most common TLR expressed during active disease, mainly by macrophages although without correlation with the amount of cytokines and number of cells.
- Published
- 2012
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39. Revisiting Langerhans cells in paracoccidioidomycosis: expression of CD207/langerin in human cutaneous and mucosal lesions.
- Author
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Pagliari C, Fernandes ER, Ferreira da Silva WL, Alves de Lima Silva A, Stegun FW, Duarte MI, and Sotto MN
- Subjects
- Animals, Biopsy, Cell Movement immunology, Dendrites, Dermis immunology, Dermis pathology, Epidermis immunology, Epidermis pathology, Granuloma immunology, Granuloma pathology, Humans, Immunohistochemistry, Inflammation pathology, Langerhans Cells immunology, Langerhans Cells pathology, Latin America, Mice, Mucous Membrane immunology, Mucous Membrane pathology, Paracoccidioides physiology, Paracoccidioidomycosis microbiology, Rabbits, Skin Diseases immunology, Skin Diseases pathology, Antigens, CD metabolism, Langerhans Cells metabolism, Lectins, C-Type metabolism, Mannose-Binding Lectins metabolism, Paracoccidioidomycosis immunology, Paracoccidioidomycosis pathology
- Abstract
Langerhans cells are identified by the expression of langerin. We detected this molecule in cutaneous and mucosal lesions in paracoccidioidomycosis, an important infection in Latin America. Langerin+ cells were scarcely distributed, with short dendrites in epidermis and epithelium and were frequent in the dermis and corium, in the inflammatory infiltrate and granulomas. Mucosal lesions presented a higher expression of langerin in lesions with loose granulomas. For the first time we presented the expression of langerin in paracoccidioidomycosis. Positive cells in dermis and corium could represent migrating Langerhans cells or a new subset of langerin+ cells with a role in paracoccidioidomycosis., (Copyright © 2011 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.)
- Published
- 2011
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40. Paracoccidioidomycosis: cells expressing IL17 and Foxp3 in cutaneous and mucosal lesions.
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Pagliari C, Fernandes ER, Stegun FW, da Silva WL, Seixas Duarte MI, and Sotto MN
- Subjects
- Biopsy, Granuloma pathology, Humans, Immunohistochemistry, Inflammation pathology, Mucous Membrane immunology, Paracoccidioidomycosis immunology, Skin immunology, Forkhead Transcription Factors analysis, Interleukin-17 analysis, Lymphocytes chemistry, Lymphocytes immunology, Mucous Membrane pathology, Paracoccidioidomycosis pathology, Skin pathology
- Abstract
We demonstrated and quantified by immunohistochemistry the population of cells expressing IL17 and Foxp3 in cutaneous and mucosal paracoccidioidomycosis lesions, associating these populations of cells with different presentations of granulomatous response. For this purpose, 61 skin biopsies and 55 oral mucosal biopsies were evaluated. Cells expressing IL17 were distributed in the inflammatory infiltrate in both groups of lesions and were found in the vessels' wall too. Foxp3+ expression was limited to the nuclei of lymphocytes in the inflammatory infiltrate. The distribution of IL17 was similar among the groups; however, Foxp3+ cells were increased in mucosal lesions that displayed compact granulomas. The results suggest that IL17 seems to play a role in paracoccidioidomycosis cutaneous and mucosal lesions, probably as secondary cells in the clearance of the fungal antigens. The presence of Foxp3+ cells both in skin and mucosa corroborates some previous researches that suggest the role of this group of cells in the modulation of local immune response., (Copyright © 2011 Elsevier Ltd. All rights reserved.)
- Published
- 2011
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41. In situ apoptosis of adaptive immune cells and the cellular escape of rabies virus in CNS from patients with human rabies transmitted by Desmodus rotundus.
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Fernandes ER, de Andrade HF Jr, Lancellotti CL, Quaresma JA, Demachki S, da Costa Vasconcelos PF, and Duarte MI
- Subjects
- Adolescent, Adult, Animals, Bites and Stings, Child, Encephalitis, Viral pathology, Encephalitis, Viral virology, Female, Humans, Male, Middle Aged, Neurons immunology, Neurons pathology, Rabies mortality, Rabies transmission, Apoptosis immunology, Central Nervous System Viral Diseases pathology, Central Nervous System Viral Diseases virology, Chiroptera virology, Lymphocytes immunology, Lymphocytes pathology, Rabies immunology, Rabies virology, Rabies virus immunology
- Abstract
The aim of the current study was to investigate the apoptosis of neurons, astrocytes and immune cells from human patients that were infected with rabies virus by vampire bats bite. Apoptotic neurons were identified by their morphology and immune cells were identified using double immunostaining. There were very few apoptotic neurons present in infected tissue samples, but there was an increase of apoptotic infiltrating CD4+ and TCD8+ adaptive immune cells in the rabies infected tissue. No apoptosis was present in NK, macrophage and astrocytes. The dissemination of the human rabies virus within an infected host may be mediated by viral escape of the virus from an infected cell and may involve an anti-apoptotic mechanism, which does not kill the neuron or pro-apoptosis of TCD4+ and TCD8+ lymphocytes and which allows for increased proliferation of the virus within the CNS by attenuation of the adaptive immune response., (Copyright © 2011 Elsevier B.V. All rights reserved.)
- Published
- 2011
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42. The expression of TLR9 in human cutaneous leishmaniasis is associated with granuloma.
- Author
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Tuon FF, Fernandes ER, Pagliari C, Duarte MI, and Amato VS
- Subjects
- Dermis immunology, Dermis pathology, Epidermis immunology, Epidermis pathology, Gene Expression Profiling, Humans, Immunohistochemistry, Macrophages immunology, Granuloma parasitology, Granuloma pathology, Leishmania braziliensis immunology, Leishmania braziliensis pathogenicity, Leishmaniasis, Cutaneous immunology, Leishmaniasis, Cutaneous pathology, Toll-Like Receptor 9 biosynthesis
- Abstract
The Toll-like receptor (TLR) signalling pathway is the first system that defends against Leishmania. After recognising Leishmania as nonself, TLRs trigger NF-κB expression.NF-κB proceeds to the nucleus and promotes the transcription of pro-inflammatory cytokines. TLR9 is thus an important factor in the induction of an effective immune response against Leishmania. We examined the pattern of TLR9 expression in 12 patients with cutaneous leishmaniasis caused by Leishmania braziliensis detected by polymerase chain reaction. Normal skin was analysed as a negative control. TLR9 expression was examined in the dermis and epidermis by immunohistochemical analysis of paraffin-embedded biopsy tissue. TLR9 expression was primarily observed in the granuloma. The protein was detected in a few cells in the dermis. A lower expression level was detected in the epidermis of patients with leishmaniasis when compared with normal skin. The presence of TLR9 in the skin of patients with cutaneous leishmaniasis is associated with granuloma and expressed by macrophages.
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- 2010
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43. The expression of TLR2, TLR4 and TLR9 in the epidermis of patients with cutaneous leishmaniasis.
- Author
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Tuon FF, Fernandes ER, Duarte MI, and Amato VS
- Subjects
- Adolescent, Adult, Epidermis pathology, Humans, Leishmaniasis, Cutaneous pathology, Male, Middle Aged, Prospective Studies, Toll-Like Receptor 2 analysis, Toll-Like Receptor 4 analysis, Toll-Like Receptor 9 analysis, Young Adult, Epidermis immunology, Leishmaniasis, Cutaneous immunology, Toll-Like Receptor 2 biosynthesis, Toll-Like Receptor 4 biosynthesis, Toll-Like Receptor 9 biosynthesis
- Published
- 2010
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44. Immunohistochemistry and polymerase chain reaction on paraffin-embedded material improve the diagnosis of cutaneous leishmaniasis in the Amazon region.
- Author
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Amato VS, Tuon FF, de Andrade HF Jr, Bacha H, Pagliari C, Fernandes ER, Duarte MI, Neto VA, Zampieri RA, Floeter-Winter LM, Celeste BJ, Oliveira J, Quiroga MM, Mascheretti M, and Boulos M
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Brazil, Humans, Immunohistochemistry, Middle Aged, Paraffin Embedding, Prospective Studies, Sensitivity and Specificity, Young Adult, Leishmaniasis, Cutaneous diagnosis, Polymerase Chain Reaction
- Abstract
Background: Recently, there has been an increase in the incidence of cutaneous leishmaniasis (CL), which represents an important health problem. This increase may be related to the epidemiologic expansion of the infective agent and the increase in tourism in tropical areas. The difficulty in clinical diagnosis, mainly in areas in which CL is not the first consideration of local physicians, has intensified efforts to describe diagnostic tests, which should be specific, sensitive, and practical. Amongst the new tests described are those including nucleic acid amplification (polymerase chain reaction, PCR) and immunohistochemistry (IHC)., Methods: In this study, we evaluated the sensitivity of a PCR based on small subunit (SSU) ribosomal DNA, in comparison with IHC using Leishmania spp. antibodies, in biopsies embedded in paraffin., Result: The results indicated a total sensitivity of 96% (90.9% with PCR and 68.8% with IHC), showing the possibility of using paraffin-embedded biopsies to diagnose CL., Conclusion: We propose the use of the two tests together as a routine protocol for diagnosis. This would require the provision of local medical services to perform molecular biology techniques and adequate Leishmania antibodies.
- Published
- 2009
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45. Wave expansion of CD34+ progenitor cells in the spleen in rodent malaria.
- Author
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Hermida FP, Vieira DP, Fernandes ER, and de Andrade HF Jr
- Subjects
- Animals, Female, Flow Cytometry, Immunohistochemistry, Mice, Mice, Inbred C57BL, Parasitemia immunology, Spleen immunology, Spleen pathology, Stem Cells cytology, Stem Cells immunology, Antigens, CD34 analysis, Malaria immunology, Plasmodium berghei immunology, Plasmodium chabaudi immunology, Spleen cytology
- Abstract
Defense against malaria depends upon amplification of the spleen structure and function for the clearance of parasitized red blood cells (pRBC). We studied the distribution and amount of CD34(+) cells in the spleens of mice infected with rodent malaria. We sought to identify these cells in the spleen and determine their relationship to infection. C57BL/6J mice were infected with self-resolving, Plasmodium chabaudi CR, or one of the lethal rodent malaria strains, P. chabaudi AJ and P. berghei ANKA. We then recorded parasitemia, mortality, and the presence of CD34(+) cells in spleen, as determined by immunohistochemistry and flow cytometry. In the non-lethal strain, the spleen structure was maintained during amplification, but disrupted in lethal models. The abundance of CD34(+) cells increased in the red pulp on the 4th and 6th days p.i. in all models, and subsided on the 8th day p.i. Faint CD34(+) staining on the 8th day p.i., was probably due to differentiation of committed cell lineages. In this work, increase of spleen CD34(+) cells did not correlate with infection control.
- Published
- 2009
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46. In situ immune responses to interstitial pneumonitis in human visceral leishmaniasis.
- Author
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Tuon FF, Guedes F, Fernandes ER, Pagliari C, Amato VS, and Seixas Duarte MI
- Subjects
- CD8-Positive T-Lymphocytes immunology, Cell Proliferation, Humans, Immunohistochemistry, Interferon-gamma biosynthesis, Interleukin-4 biosynthesis, Leishmaniasis, Visceral complications, Lung Diseases, Interstitial parasitology, Lung Diseases, Parasitic etiology, Macrophages immunology, Tumor Necrosis Factor-alpha biosynthesis, Leishmaniasis, Visceral immunology, Lung Diseases, Interstitial immunology, Lung Diseases, Parasitic immunology
- Abstract
Lung disease during active human visceral leishmaniasis is frequently reported. As such, studies have associated pulmonary symptoms to interstitial pneumonitis with a mononuclear infiltrate. However, the immune response in this condition has never been described before. The aim of this study was to determine the immunophenotypic pattern and cytokine profile of lung involvement (IPL) in human visceral leishmaniasis. Quantitative methods of analysis were performed using immunohistochemistry, and were compared with a control group of normal lung. Interstitial macrophages and cd8 cells were increased in IPL, and IL-4 as well as TNF-alpha displayed increased expression when compared to the control group. This inflammatory process with a Th2 pattern, as suggested by increased IL-4 and low IFN-gamma expression, is consistent with the immune response in other organs of visceral leishmaniasis. The microenvironment of the immune response in this condition is associated with lung disease in patients with interstitial pneumonitis related to visceral leishmaniasis, increasing the chance of bacterial infection.
- Published
- 2009
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47. Association between 894G>T endothelial nitric oxide synthase gene polymorphisms and metabolic syndrome.
- Author
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Piccoli JC, Gottlieb MG, Castro L, Bodanese LC, Manenti ER, Bogo MR, Peres A, Rocha MI, and Cruz IB
- Subjects
- Adult, Aged, Aged, 80 and over, Epidemiologic Methods, Female, Genotype, Humans, Hypertension genetics, Male, Middle Aged, Metabolic Syndrome genetics, Nitric Oxide Synthase Type III genetics, Polymorphism, Genetic genetics
- Abstract
Metabolic syndrome (MS) is a cluster of cardiovascular risk factors such as hypertension, dyslipidemia, obesity and type II diabetes. Here, we performed a case-control study analyzing the association between 894G>T endothelial nitric oxide synthase gene polymorphism (NOS3) and MS in 616 subjects. Genotype frequencies were TT= 9.3%, GG= 37.2 and TG= 53.6% and the allelic frequencies were T=0.36 and G= 0.64. We observed a higher TT genotype frequency in the male MS group than control subjects (p=0.02), independent of other variables. We found an association between hypertension and TT genotype in females. Our data suggests that 894G>T plays a significant role in the mechanistic interaction between metabolic risk such as hypertension and MS, although sex-related differences may exist.
- Published
- 2008
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48. The effects of human herpesvirus 8 infection and interferon-gamma response in cutaneous lesions of Kaposi sarcoma differ among human immunodeficiency virus-infected and uninfected individuals.
- Author
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Guedes F, de Andrade HF Jr, Fernandes ER, Tuon FF, Brasil RA, Pagliari C, and Duarte MI
- Subjects
- AIDS-Related Opportunistic Infections virology, Acquired Immunodeficiency Syndrome drug therapy, Acquired Immunodeficiency Syndrome immunology, Aged, Aged, 80 and over, Antigens, Viral metabolism, Antiretroviral Therapy, Highly Active, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Female, Humans, Immunity, Cellular, Interferon-gamma metabolism, Male, Middle Aged, Nuclear Proteins metabolism, Sarcoma, Kaposi drug therapy, Sarcoma, Kaposi virology, Skin Neoplasms drug therapy, Skin Neoplasms virology, AIDS-Related Opportunistic Infections immunology, Herpesvirus 8, Human immunology, Sarcoma, Kaposi immunology, Skin Neoplasms immunology
- Abstract
Background: Kaposi sarcoma (KS) is associated with human herpesvirus 8 (HHV-8). The cutaneous immune response in this tumour is not well established and a better understanding is necessary., Objectives: To evaluate the HHV-8 expression and immune response in cutaneous lesions of classic KS (CKS) and AIDS-associated KS (AIDS-KS)., Methods: We performed a quantitative immunohistochemical study of cells expressing HHV-8 latency-associated nuclear antigen (LANA), CD4, CD8 and interferon (IFN)-gamma in skin lesions from patients with CKS and AIDS-KS (with or without highly active antiretroviral therapy, HAART)., Results: CKS showed higher LANA expression compared with AIDS-KS, regardless of HAART. We also found higher LANA expression in nodules compared with patch/plaque lesions. The tissue CD4+ cell proportion was lower in AIDS-KS patients without HAART than in patients with CKS. In CKS lesions, CD4+ and CD8+ cells expressed IFN-gamma, as shown by double immunostaining. AIDS-KS presented low numbers of IFN-gamma-expressing cells. CD8+ cell numbers were similar in all groups, which appeared unrelated to the clinical or epidemiological type of KS., Conclusions: Our quantitative data on the pattern of KS lesions in selected groups of patients, as shown by in situ immune response, demonstrated a CD4+ T-cell involvement associated with IFN-gamma, an environment of immune response-modified human immunodeficiency virus (HIV) infection. In our sample, the promotion of KS in patients without HIV appears to be related to higher HHV-8 load or virulence than in those with AIDS. This higher resistance may be explained by a sustained immune response against this herpesvirus, that is only partially restored but effective after HAART.
- Published
- 2008
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49. Interaction of human papillomavirus DNA with factor XIIIa-positive dermal dendrocytes in vulvar lesions.
- Author
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Pereira NV, Pagliari C, Fernandes ER, Guedes F, Sotto MN, and Duarte MI
- Subjects
- Female, Humans, Vulvar Diseases immunology, DNA, Viral immunology, Dendritic Cells immunology, Dermis cytology, Factor XIIIa analysis, Papillomaviridae genetics, Papillomavirus Infections immunology
- Published
- 2008
- Full Text
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50. Portal CD4+ and CD8+ T lymphocyte correlate to intensity of interface hepatitis in chronic hepatitis C.
- Author
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Viso AT, de Castro Barbosa T, Yamamoto L, Pagliari C, Fernandes ER, Brasil RA, Andrade HF Jr, Duarte MI, and Barone AA
- Subjects
- Adolescent, Adult, Disease Progression, Female, Genotype, Hepacivirus genetics, Hepatitis C, Chronic pathology, Humans, Immunohistochemistry, Liver blood supply, Liver pathology, Male, Middle Aged, Severity of Illness Index, CD4-CD8 Ratio, Hepacivirus immunology, Hepatitis C, Chronic immunology, Liver virology
- Abstract
Background: The pathogenesis of chronic hepatitis C is still a matter of debate. CD4+ and CD8+ T lymphocytes (TL) are typically observed within the portal and periportal spaces of affected livers, but their functional role in hepatitis C progression has not been fully elucidated., Methods: CD4+ and CD8+ TL were quantified by immunohistochemistry in portal and periportal spaces of 39 liver biopsies from patients with chronic hepatitis C. They were associated to demographic data, histological parameters, laboratory findings of patients and hepatitis C genotypes., Results: There was high numbers of CD4+ and CD8+ TL from which the density of CD4+ T was higher than CD8+ TL in portal and periportal spaces. CD4+ and CD8+ TL were directly correlated to intensity of interface hepatitis. CD8+ TL correlated to serum enzyme levels., Conclusion: The high numbers of CD4+ and CD8+ TL in portal and periportal spaces and their correlation to interface hepatitis suggest that hepatitis C evolution depends on the action of intrahepatic T lymphocytes, lending support to the notion of an immune-mediated mechanism in the pathogenesis of chronic hepatitis C.
- Published
- 2007
- Full Text
- View/download PDF
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