Cinética de carga viral em componentes do sangue de pacientes cronicamente infectados pelo vírus da hepatite C, antes e durante terapia tripla.

Submitted by Nuzia Santos ([email protected]) on 2017-12-20T18:15:02Z No. of bitstreams: 1 Dissertacao_BCM_Jordana Rodrigues Barbosa Fradico - Cinética de carga viral em componentes do sangue de pacientes cronicamente infectados pelo vírus da hepatite C, antes e durante terapia tripla - 2017.pdf: 2989814 bytes, checksum: c2766b20ebc2a64706e7066b53f8dc90 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2017-12-20T18:25:58Z (GMT) No. of bitstreams: 1 Dissertacao_BCM_Jordana Rodrigues Barbosa Fradico - Cinética de carga viral em componentes do sangue de pacientes cronicamente infectados pelo vírus da hepatite C, antes e durante terapia tripla - 2017.pdf: 2989814 bytes, checksum: c2766b20ebc2a64706e7066b53f8dc90 (MD5)Made available in DSpace on 2017-12-20T18:25:58Z (GMT). No. of bitstreams: 1 Dissertacao_BCM_Jordana Rodrigues Barbosa Fradico - Cinética de carga viral em componentes do sangue de pacientes cronicamente infectados pelo vírus da hepatite C, antes e durante terapia tripla - 2017.pdf: 2989814 bytes, checksum: c2766b20ebc2a64706e7066b53f8dc90 (MD5) Previous issue date: 2017Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.A hepatite C é um grave problema de saúde pública mun dial, causada pelo vírus da hepatite C (VHC). Apesar dos hepatócitos serem o alvo principal do VHC, o vírus pode ainda interagir com monócitos, linfócitos, célu las endoteliais, eritrócitos e plaquetas. Neste contexto o presente estudo buscou a valiar a cinética viral em componentes do sangue de pacientes sintomáticos cron icamente infectados pelo VHC, antes e durante a terapia tripla (interferon-pe guilado/ribavirina e inibidor de protease). Com esta finalidade, os seguintes objetivos específi cos foram propostos para a população avaliada: otimizar uma metodologia in house para adaptação do kit Abbott RealTime HCV ® (Abbott Molecular, EUA) para a execução da quantifica ção da carga viral de VHC em soro, sangue total, eritrócit os, plaquetas e leucócitos; quantificar a carga viral de VHC nesses componentes sa nguíneos; associar e correlacionar a carga viral de VHC com dados laborato riais dos pacientes antes e durante terapia tripla. Os objetivos foram cumprido s pela execução da metodologia de transcrição reversa seguida de PCR em tempo real ( RT-qPCR) utilizando o kit Abbott RealTime HCV ® . Dessa maneira avaliou-se amostras de componentes sanguíneos de 20 pacientes com diagnóstico confirmado de hepatite C crônica, infectados pelo VHC genótipo 1. Esta população de estu do apresentou perfis epidemiológicos e clínico-laboratoriais semelhantes aos descritos na literatura. Obteve-se otimização do kit Abbott RealTime HCV ® para quantificação do RNA-VHC em condições de pesquisa e aumentou-se em 4 vezes o número de reações por kit, resultando em uma economia financeira considerável. A análise da quantificação da carga viral foi realizada por quatro variáveis, sendo elas: tempo de tratamento, componente sanguíneo, níveis da carga viral e desfech o clínico. Em todas, observou-se que o soro foi o componente com maiores níveis de carga viral e as hemácias com menor nível. Durante a terapia tripla, houve redução dos níveis de cargas virais em todos os componentes sanguíneos anal isados. Esta redução só não foi observada na 24ª semana de tratamento, quand o houve aumento das cargas virais, quando comparado com a quarta semana. Estes ní veis apresentaram queda na 48ª semana sem, no entanto, atingir carga nula no soro, sangue total, hemácias e plaquetas. Positividade que não foi capaz de alterar o desfecho clínico dos pacientes, que se mantiveram apresentando resposta v irológica sustentada (RVS). A concretização deste estudo demonstrou que pacientes infectados pelo VHC cronicamente apresentaram altas cargas de VHC associa das aos componentes sanguíneos. O tratamento medicamentoso da infecção a presentou-se eficiente e efetivo, provocando uma alteração na cinética de in teração do vírus com os componentes sanguíneos. O aumento das cargas virais n a 24ª semana é um indicativo de que esta é uma semana crítica na terap ia tripla, que merece maior atenção dos profissionais de saúde. A constatação de baixas cargas virais no soro, sangue total, hemácias e plaquetas na 48ª semana da t erapia foi um resultado surpreendente do estudo, visto que 85,7% dos pacient es obtiveram RVS. Sugere-se a continuidade de estudos que aprofundem pontos impo rtantes evidenciados por este trabalho. Os resultados obtidos proporcionaram um entendimento maior da fisiopatologia do processo e da interação do VHC com os componentes do sangue.ABSTRACT Hepatitis C is a serious global public health proble m, caused by the hepatitis C virus (HCV). Although hepatocytes are the major target of HCV, the virus may still interact with monocytes, lymphocytes, endothelial cells, eryt hrocytes, and platelets. In this context, the present study sought to evaluate viral kinetics in blood components of patients chronically infected with HCV before and dur ing triple therapy (pegylated interferon/ ribavirin and protease inhibitor). The following specific objectives were proposed: to optimize an in-house methodology for th e adaptation of the Abbott RealTime HCV ® kit (Abbott Molecular, USA) for the quantification of HCV viral load in serum, whole blood, erythrocytes, platelets and leuk ocytes; to quantify the HCV load in these blood components; to associate and correla te HCV load with laboratorial data of patients before and during triple therapy. T he objectives were met by the implementation of the reverse transcription methodo logy followed by real-time PCR (RT-qPCR) using the Abbott RealTime HCV ® kit. Using this approach, peripheral blood samples from 20 patients with a confirmed diagn osis of chronic hepatitis C, infected by HCV genotype 1, were analyzed. This study population presented epidemiological and clinical-laboratorial profiles s imilar to those described in the literature. Abbott RealTime HCV ® kit was optimized for HCV RNA quantification in research conditions and increased the number of rea ctions per kit in 4 times. The analysis of the viral load was performed by four var iables: treatment time, blood component, viral load levels and clinical outcome. In all of them, it was observed that serum was the component with the highest levels of vir al load and the red cells with the lowest level. During triple therapy, there was a r eduction in levels of viral loads in all blood components analyzed. This reduction was no t observed at 24th week of treatment, when viral loads increased when compared t o the fourth week. At 48th week, the viral load levels were decreased without, ho wever, reaching zero load in serum, whole blood, red blood cells and platelets. T his persistent positivity was not able to change the clinical outcome of the patients, who maintained a sustained virological response (SVR). The achievement of this st udy demonstrated that HCV chronically infected patients present high HCV loads associated with blood components. The drug treatment of the infection was e fficient and effective, causing a change in the kinetics of virus interaction in the blood components. The increase in viral load at 24th week is indicative that this is a critical week in triple therapy, which deserves more attention from health professionals. Low viral load in serum, whole blood, red blood cells and platelets at 48th week o f therapy was a surprising result of the study, since 85.7% of the patients obtained SVR. This finding suggests that additional studies could im prove the understanding of the mechanisms responsible for the blood components-HCV interaction during the viral infection

Structural Investigation of Anti-Trypanosoma cruzi 2-Iminothiazolidin-4-ones Allows the Identification of Agents with Efficacy in Infected Mice

We modified the thiazolidinic ring at positions N3, C4, and C5, yielding compounds 6-24. Compounds with a phenyl at position N3, 15-19, 22-24, exhibited better inhibitory properties for cruzain and against the parasite than 2-iminothiazolidin-4-one S. We were able to identify one high-efficacy trypanocidal compound, 2-minothiazolidin-4-one 18, which inhibited the activity of cruzain and the proliferation of epirnastigotes and was cidal for trypomastigotes but was not toxic for splenocytes. Having located some of the structural determinants of the trypanocidal properties, we subsequently wished to determine if the exchange of the thiazolidine for a thiazole ring leaves the functional properties unaffected. We therefore tested thiazoles 26-45 and observed that they did not inhibit cruzain, but they exhibited trypanocidal effects. Parasite development was severely impaired when treated with 18, thus reinforcing the notion that this class of heterocycles can lead to useful cidal agents for Chagas disease.CNPq [471461/2011-3]CAPES [23038.003155/2011-37]FAPESB (PRONEX grant)European Union ChemBioFight [269301]CAPES-Fulbright Foundatio

Acute-Phase Levels of CXCL8 as Risk Factor for Chronic Arthralgia Following Chikungunya Virus Infection

The immunopathogenesis of chikungunya virus (CHIKV) infection and the role of acute-phase immune response on joint pain persistence is not fully understood. We investigated the profile of serum chemokine and cytokine in CHIKV-infected patients with acute disease, compared the levels of these biomarkers to those of patients with other acute febrile diseases (OAFD) and healthy controls (HC), and evaluated their role as predictors of chronic arthralgia development. Chemokines and cytokines were measured by flow Cytometric Bead Array. Patients with CHIKV infection were further categorized according to duration of arthralgia (≤ 3 months vs >3 months), presence of anti-CHIKV IgM at acute-phase sample, and number of days of symptoms at sample collection (1 vs 2-3 vs ≥4). Patients with acute CHIKV infection had significantly higher levels of CXCL8, CCL2, CXCL9, CCL5, CXCL10, IL-1β, IL-6, IL-12, and IL-10 as compared to HC. CCL2, CCL5, and CXCL10 levels were also significantly higher in patients with CHIKV infection compared to patients with OAFD. Patients whose arthralgia lasted > 3 months had increased CXCL8 levels compared to patients whose arthralgia did not (p3 months. Patients with chikungunya and OAFD had similar cytokine kinetics for IL-1β, IL-12, TNF, IFN-γ, IL-2, and IL-4, although the levels were lower for CHIKV patients. This study suggests that chemokines may have an important role in the immunopathogenesis of chronic chikungunya-related arthralgia.Fil: Jacob Nascimento, Leile Camila. Fundación Oswaldo Cruz; BrasilFil: Carvalho, Caroline Xavier. Fundación Oswaldo Cruz; BrasilFil: Oliveira Silva, Monaíse Madalena. Fundación Oswaldo Cruz; BrasilFil: Kikuti, Mariana. Fundación Oswaldo Cruz; Brasil. Universidade Federal da Bahia; BrasilFil: Oliveira Anjos, Rosângela. Fundación Oswaldo Cruz; BrasilFil: Rodrigues Barbosa Fradico, Jordana. Fundación Oswaldo Cruz; BrasilFil: Campi Azevedo, Ana Carolina. Fundación Oswaldo Cruz; BrasilFil: Tauro, Laura Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú; ArgentinaFil: Soares Campos, Gúbio. Universidade Federal da Bahia; BrasilFil: Sousa dos Santos Moreira, Patricia. Fundación Oswaldo Cruz; BrasilFil: Machado Portilho, Moyra. Fundación Oswaldo Cruz; BrasilFil: Assis Martins Filho, Olindo. Fundación Oswaldo Cruz; BrasilFil: Sousa Ribeiro, Guilherme. Universidade Federal da Bahia; Brasil. Fundación Oswaldo Cruz; BrasilFil: Galvão Reis, Mitermayer. Fundación Oswaldo Cruz; Brasil. University of Yale; Estados Unidos. Universidade Federal da Bahia; Brasi

Duration of Humoral and Cellular Immunity 8 Years After Administration of Reduced Doses of the 17DD-Yellow Fever Vaccine

Corrigendum: In the original article, there was a mistake in Figure 5 as published. One orange frame erroneously shifted slightly to the right. The corrected Figure 5 appears below. Additionally, there was a mistake in Supplementary Figure 2 as published. The asterisks indicating statistical significance were erroneously deleted during the JPEG conversion. It is important to mention that no results have been modified. The corrected Supplementary Figure 2 appears below.Submitted by Nuzia Santos ([email protected]) on 2019-08-21T13:30:06Z No. of bitstreams: 1 Duration of Humoral and Cellular Immunity 8 Years After .pdf: 3045695 bytes, checksum: 524cbfff77a5b05dcf0f4526fae5fb3c (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2019-08-21T13:43:11Z (GMT) No. of bitstreams: 1 Duration of Humoral and Cellular Immunity 8 Years After .pdf: 3045695 bytes, checksum: 524cbfff77a5b05dcf0f4526fae5fb3c (MD5)Made available in DSpace on 2019-08-21T13:43:11Z (GMT). No. of bitstreams: 1 Duration of Humoral and Cellular Immunity 8 Years After .pdf: 3045695 bytes, checksum: 524cbfff77a5b05dcf0f4526fae5fb3c (MD5) Previous issue date: 2019Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil / Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. Departamento de Microbiologia. Laboratório de Virologia Básica e Aplicada. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Núcleo de Estudos em Saúde Pública e Envelhecimento. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Escola Nacional de Saúde Pública. Departamento de Epidemiologia e Métodos Quantitativos em Saúde. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Rio de Janeiro, RJ, Brasil.Instituto de Biologia do Exército. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Rio de Janeiro, RJ, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Departamento de Vigilância das Doenças Transmissíveis. Brasília, DF, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Departamento de Vigilância das Doenças Transmissíveis. Brasília, DF, Brasil.Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brasil.The present study aims to determine whether 17DD-YF-specific humoral and cellular immunological memory is maintained 8-years after primary vaccination with subdoses (10,447IU;3,013IU;587IU;158IU;31IU). For this purpose, this follow-up study was carried out in a subset of volunteers (n = 98) originally enrolled in the dose-response study in 2009 and 46 non-vaccinated controls. Our results demonstrated that vaccinees, who had seroconverted following primary vaccination and had not been revaccinated, present similar neutralizing antibodies levels and YF-specific cellular memory, particularly CMCD4 and EMCD8 as compared to the reference full dose (27,476IU). Although, PRNT seropositivity rates were similar across subgroups (94, 82, 83, 94, 80, and 91%, correspondingly), only doses above 587IU elicited similar iterative proportion of seropositivity rates, calculated as a progressive decrease on seropositivity rates along time (89, 80, 80, and 91%, respectively) as compared to 158IU and 31IU (68 and 46%, respectively). Noteworthy were the strong positive correlations ("EMCD4,EMCD8" and "TNFCD8,IFNCD8") observed in most subdoses, except for 31IU. Major similarities underscored the preserved antibody titers and the outstanding levels of EMCD8, relevant correlates of protection for YF-specific immunity. These findings provide evidences to support the regular use of dose sparing strategy for YF vaccine in adults

Late-Relapsing Hepatitis after Yellow Fever

One patient presented hyporexia, asthenia, adynamia, and jaundice two months after acute yellow fever (YF) onset; plus laboratory tests indicating hepatic cytolysis and a rebound of alanine and aspartate transaminases, and total and direct bilirubin levels. Laboratory tests discarded autoimmune hepatitis, inflammatory or metabolic liver disease, and new infections caused by hepatotropic agents. Anti-YFV IgM, IgG and neutralizing antibodies were detected in different times, but no viremia. A liver biopsy was collected three months after YF onset and tested positive for YFV antigens and wild-type YFV-RNA (364 RNA-copies/gram/liver). Transaminases and bilirubin levels remained elevated for five months, and the arresting of symptoms persisted for six months after the acute YF onset. Several serum chemokines, cytokines, and growth factors were measured. A similar immune response profile was observed in the earlier phases of the disease, followed by more pronounced changes in the later stages, when transaminases levels returned to normal. The results indicated viral persistence in the liver and continual liver cell damage three months after YF onset and reinforced the need for extended follow-ups of YF patients. Further studies to investigate the role of possible viral persistence and the immune response causing relapsing hepatitis following YF are also necessary

Structural Investigation of Anti-<i>Trypanosoma cruzi</i> 2‑Iminothiazolidin-4-ones Allows the Identification of Agents with Efficacy in Infected Mice

We modified the thiazolidinic ring at positions N3, C4, and C5, yielding compounds <b>6</b>–<b>24</b>. Compounds with a phenyl at position N3, <b>15</b>–<b>19</b>, <b>22</b>–<b>24</b>, exhibited better inhibitory properties for cruzain and against the parasite than 2-iminothiazolidin-4-one <b>5</b>. We were able to identify one high-efficacy trypanocidal compound, 2-minothiazolidin-4-one <b>18</b>, which inhibited the activity of cruzain and the proliferation of epimastigotes and was cidal for trypomastigotes but was not toxic for splenocytes. Having located some of the structural determinants of the trypanocidal properties, we subsequently wished to determine if the exchange of the thiazolidine for a thiazole ring leaves the functional properties unaffected. We therefore tested thiazoles <b>26</b>–<b>45</b> and observed that they did not inhibit cruzain, but they exhibited trypanocidal effects. Parasite development was severely impaired when treated with <b>18</b>, thus reinforcing the notion that this class of heterocycles can lead to useful cidal agents for Chagas disease