Estabelecimento do modelo de Hepatite crônica pelo vírus da Hepatite E (genótipo 3) em Macaca fascicularis

Submitted by Claudete Fernandes ([email protected]) on 2017-07-19T13:19:15Z No. of bitstreams: 2 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) 73873.pdf: 16908741 bytes, checksum: 2586bd46bf6ab29694ccd5dd48f76fc5 (MD5)Approved for entry into archive by Anderson Silva ([email protected]) on 2017-08-23T13:18:46Z (GMT) No. of bitstreams: 2 73873.pdf: 16908741 bytes, checksum: 2586bd46bf6ab29694ccd5dd48f76fc5 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Made available in DSpace on 2017-08-23T13:18:46Z (GMT). No. of bitstreams: 2 73873.pdf: 16908741 bytes, checksum: 2586bd46bf6ab29694ccd5dd48f76fc5 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2017Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, BrasilA infecção pelo vírus da hepatite E (HEV) é hoje considerada a principal causa de hepatite viral aguda no mundo. Os genótipos 3 e 4 (HEV-3 e HEV-4) podem infectar várias espécies animais, tais como suínos, constituindo assim um risco para a cadeia alimentar humana em muitos países. Acreditava-se que o HEV causasse apenas hepatite aguda autolimitada. No entanto, já se reconhece que a infecção pelo HEV pode evoluir para hepatite crônica em pacientes immunossuprimidos, principalmente em receptores de órgãos sólidos. A infecção crônica pelo HEV pode evoluir para hepatite crônica e fibrose dentro de dois anos em aproximadamente 60% dos pacientes que recebem tratamento imunossupressor póstransplante. O tacrolimo, medicamento de eleição para terapia imunossupressora de receptores de transplante sólidos, tem sido considerado um fator de risco potencial para o estabelecimento de uma infecção crônica pelo HEV-3. O presente estudo teve por objetivo investigar a patogênese da infecção persistente pelo HEV usando macacos cinomolgos como modelo animal para avaliar a relação entre a condição de imunossupressão do hospedeiro e a ocorrência e persistência da infecção pelo HEV com consequente evolução para hepatite crônica. Nesse estudo nós descrevemos o sucesso obtido na infecção experimental de macacos cinomolgos, previamente tratados com o imunossupressor tacrolimo, e inoculados com uma cepa brasileira do HEV-3 isolada de um suíno naturalmente infectado. Aos três meses após a infecção, três dos quatro macacos imunossuprimidos exibiram um padrão persistente de viremia, excreção viral nas fezes e presença do HEV RNA em todas as biópsias hepáticas, achados compatíveis com uma infecção crônica pelo HEV Todos os animais soroconverteram para anti-HEV (IgA, IgM e IgG). O grupo de animais imunossuprimidos apresentou títulos mais baixos de IgM e soroconversão tardia para IgG, em comparação com os animais imunocompetentes. Todos os macacos cronicamente infectados exibiram picos baixos e intermitentes das aminotransferases hepáticas, ALT e AST, enquanto os animais imunocompetentes, com infecção aguda, apresentaram apenas um discreto pico de elevação. Ao final do experimento, a replicação viral foi evidenciada pela detecção de RNA de fita positiva e de fita negativa no tecido hepático, e pela presença do antígeno HEV nos hepatócitos e sinusóides. As amostras de tecido hepático obtidas na necropsia exibiram inflamação de interface. Nossos achados forneceram evidências de que macacos cinomolgos tratados com o imunossupressor tacrolimo e inoculados com a cepa brasileira do HEV-3 suíno (Genebank: KX578263/KX578267), foram persistentemente infectados. A infecção pelo HEV evoluiu com hepatite de interface, achado característico de hepatite crônicaHepatitis E virus (HEV) infection is a major cause of acute viral hepatitis worldwide. HEV genotypes 3 and 4 can infect a variety of animal species, such pigs, and can be found in the human food chain of a number in many countries. HEV was believed to cause only selflimiting acute hepatitis. However, it is now accepted that HEV may lead to chronic hepatitis in immunosuppressed patients, mainly in solid organ transplant (SOT) recipients. Chronic HEV infection may evolve to chronic hepatitis and fibrosis within the first two years of infection in approximately 60% of the patients receiving immunosuppressive treatment following solid-organ transplantation (SOT). Tacrolimus, the immunosuppressive therapy of choice for treatment of SOT recipients, has been considered a potential risk factor related to HEV-3 chronic infection. The present study aimed to investigate the pathogenesis of HEV persistent infection by using cynomolgus monkeys as an animal model for evaluating the relationship between the host immunosuppressive status and the occurrence and persistence of a HEV infection leading to chronic hepatitis. Here we describe a successful experimental study to investigate the pathogenesis of HEV chronic infection by using cynomolgus monkeys, previously immunosuppressed with tacrolimus, and inoculated with a Brazilian HEV-3 strain isolated from a naturally infected pig. Three months post infection three out of four immunosuppressed monkeys exhibited a persistent pattern of viremia, faecal shedding and presence of HEV RNA in all liver biopsies, that is compatible with a chronic HEV infection. All animals seroconverted to anti-HEV (IgA, IgM and IgG) The immunosuppressed group showed lower IgM titres, and later IgG seroconversion in comparison with immunocompetent animals. Of note, all chronically infected monkeys exhibited low and intermittent peaks of ALT and AST levels while the immunocompetent, acutely infected monkeys showed only a discrete elevation peak. At the end of the experiment, viral replication was evidenced by the detection of positive- and negativestranded RNA in the liver tissue, and by the presence of HEV antigen in hepatocytes and hepatic sinusoids. Liver sections obtained at necropsy showed interface inflammation. Our findings provided the evidence that immunosuppressed cynomolgus monkeys treated with tacrolimus and inoculated with swine HEV-3 (Genebank: KX578263/KX578267) can be persistently infected. HEV infection was associated with interface hepatitis, which may evolve to the beginning of a chronic hepatiti

Hepatitis E: Update on Prevention and Control

Submitted by Sandra Infurna ([email protected]) on 2018-06-05T13:21:35Z No. of bitstreams: 1 juliana_melgaço_etal_IOC_2018.pdf: 1774103 bytes, checksum: d9b10681ef0334aaab9eece71880f6d1 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2018-06-05T13:28:26Z (GMT) No. of bitstreams: 1 juliana_melgaço_etal_IOC_2018.pdf: 1774103 bytes, checksum: d9b10681ef0334aaab9eece71880f6d1 (MD5)Made available in DSpace on 2018-06-05T13:28:26Z (GMT). No. of bitstreams: 1 juliana_melgaço_etal_IOC_2018.pdf: 1774103 bytes, checksum: d9b10681ef0334aaab9eece71880f6d1 (MD5) Previous issue date: 2018Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Ambulatório. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz.Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Ambulatório. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Ambulatório. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Ambulatório. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Ambulatório. Rio de Janeiro, RJ. Brasil.Hepatitis E virus (HEV) is a common etiology of acute viral hepatitis worldwide. Recombinant HEV vaccines have been developed, but only one is commercially available and licensed in China since 2011. Epidemiological studies have identified genotype 3 as the major cause of chronic infection in immunocompromised individuals. Ribavirin has been shown to be effective as a monotherapy to induce HEV clearance in chronic patients who have undergone solid organ transplant (SOT) under immunosuppressive therapy. Efforts and improvements in prevention and control have been made to reduce the instances of acute and chronic hepatitis E in endemic and nonendemic countries. However, this review shows that further studies are required to demonstrate the importance of preventive vaccination and treatment worldwide, with emphasis on hepatitis E infection in the public health system

Production and characterization of a Brazilian candidate antigen for Hepatitis E Virus genotype 3 diagnosis.

Hepatitis E, caused by hepatitis E virus (HEV), is a viral infectious pathology of great importance in the public health. Hepatitis E outbreaks were registered in developing countries with poor or no sanitation, where drinking water was contaminated with fecal material, but also in many industrialized countries probably due to consumption of HEV-positive swine meat. In this study, we present the development and characterization of a recombinant antigen from ORF2 HEV genotype 3. Viral RNA was extracted from swine feces infected with the native virus. A total of 267 residues from the C-terminal ORF2((394-661)) coding sequence were cloned into the pET20a vector and expressed in Escherichia coli ER2566. Recombinant protein was purified by liquid chromatography and the fragment obtained a 98% homology against other human or swine HEV genotype 3 ORF2 sequences. Wistar rats were inoculated with ORF2p, developing antibodies able to recognize both the homologous antigen and the native HEV genotype 3 ORF2 present in infected stool. In parallel, HEV-negative swine were experimentally challenged with HEV genotype 3. ORF2 was detected by PCR 14 days post-inoculation in three-fourth piglets' feces and one week later by dot blot. In conclusion, this study proved the immunogenic and antigenic properties of the recombinant protein ORF2p.info:eu-repo/semantics/publishe

Cynomolgus monkeys (Macaca fascicularis) experimentally and naturally infected with hepatitis E virus: The bone marrow as a possible new viral target.

Hepatitis E virus (HEV) transmission through infected blood and blood products has already been described. However, little is known about the bone marrow (BM) as source of HEV infection. Our study aimed to investigate the presence of HEV antigen (Ag) and histological changes in BM of cynomolgus monkeys (Macaca fascicularis) experimentally and naturally infected with HEV. Four cynomolgus monkeys with acute, and two with chronic hepatitis E ─ after immunosuppressive therapy with tacrolimus ─ were compared with one colony-bred animal naturally infected. Both, natural and experimental infections were characterized by anti-HEV IgG seroconversion detected by ELISA, and viral RNA isolation confirmed by RT-qPCR and qualitative nested RT-PCR. BM biopsies were collected from all animals, submitted to histology and indirect immunofluorescence techniques and observed, respectively, by light and confocal microscopy. The HEV Ag-fluorescent-labeled cells were detected from BM biopsies obtained from three monkeys with acute and one with chronic hepatitis E, and also from the naturally infected monkey. In the experimentally infected animals with acute hepatitis, HEV Ag detection occurred at 160 days post-infection, even after viral clearance in serum, feces, and liver. Double-stranded RNA, a replicative marker, was detected in BM cells from both acute and chronically infected animals. Major histological findings included vacuolization in mononuclear and endosteal cells, an absence of organized inflammatory infiltrates, and also some fields suggesting displasic focal BM disease. These findings support the hypothesis of BM cells as secondary target sites of HEV persistence. Further experimental studies should be carried out to confirm the assumption of HEV transmission through BM transplantation

Hepatitis E seroprevalence and associated factors in rural settlers in Central Brazil

Abstract INTRODUCTION: Prevalence of hepatitis E virus (HEV) infection and associated factors were investigated in rural settlements in Central Brazil. METHODS: A total of 464 settlers were interviewed, and serum samples were tested for anti-HEV IgG/IgM. Positive samples were tested for HEV RNA. RESULTS: Sixteen participants (3.4%; 95% CI 2.0-5.7) were positive for anti-HEV IgG. None was positive for anti-HEV IgM. HEV RNA was not detected. Dwelling in a rural settlement for >5 years was associated with HEV seropositivity. CONCLUSIONS: The results revealed the absence of acute infection and a low prevalence of previous exposure to HEV

Hepatitis E virus infection in patients with acute non-A, non-B, non-C hepatitis in Central Brazil

Hepatitis E virus (HEV) infection has a worldwide distribution and represents an important cause of acute hepatitis. This study aims to investigate the occurrence of HEV infection and factors associated with this infection in patients with acute non-A, non-B, non-C hepatitis in Central Brazil. From April 2012 to October 2014, a cross-sectional study was conducted among 379 patients with acute non-A, non-B, non-C hepatitis in the City of Goiania, Central Brazil. Serum samples of all patients were tested for serological markers of HEV infection (anti-HEV IgM and IgG) by ELISA. Positive samples were confirmed using immunoblot test. Anti-HEV IgM and IgG positive samples were tested for HEV RNA. Of the 379 serum samples, one (0.3%) and 20 (5.3%) were positive for anti-HEV IgM and IgG, respectively. HEV RNA was not found in any sample positive for IgM and/or IgG anti-HEV. After multivariate analysis, low education level was independently associated with HEV seropositivity (p = 0.005), as well as living in rural area, with a borderline p-value (p = 0.056). In conclusion, HEV may be responsible for sporadic self-limited cases of acute hepatitis in Central Brazil

Hepatitis E virus infection in patients with acute non-A, non-B, non-C hepatitis in Central Brazil

Hepatitis E virus (HEV) infection has a worldwide distribution and represents an important cause of acute hepatitis. This study aims to investigate the occurrence of HEV infection and factors associated with this infection in patients with acute non-A, non-B, non-C hepatitis in Central Brazil. From April 2012 to October 2014, a cross-sectional study was conducted among 379 patients with acute non-A, non-B, non-C hepatitis in the City of Goiania, Central Brazil. Serum samples of all patients were tested for serological markers of HEV infection (anti-HEV IgM and IgG) by ELISA. Positive samples were confirmed using immunoblot test. Anti-HEV IgM and IgG positive samples were tested for HEV RNA. Of the 379 serum samples, one (0.3%) and 20 (5.3%) were positive for anti-HEV IgM and IgG, respectively. HEV RNA was not found in any sample positive for IgM and/or IgG anti-HEV. After multivariate analysis, low education level was independently associated with HEV seropositivity (p = 0.005), as well as living in rural area, with a borderline p-value (p = 0.056). In conclusion, HEV may be responsible for sporadic self-limited cases of acute hepatitis in Central Brazil

Cynomolgus Monkeys (Macaca fascicularis) as an Experimental Infection Model for Human Group A Rotavirus

Submitted by Sandra Infurna ([email protected]) on 2018-10-09T13:42:57Z No. of bitstreams: 1 gentil_bentes_etal_IOC_2018.pdf: 3661954 bytes, checksum: 45163caeab19a76f53c50788c263f240 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2018-10-09T13:52:24Z (GMT) No. of bitstreams: 1 gentil_bentes_etal_IOC_2018.pdf: 3661954 bytes, checksum: 45163caeab19a76f53c50788c263f240 (MD5)Made available in DSpace on 2018-10-09T13:52:24Z (GMT). No. of bitstreams: 1 gentil_bentes_etal_IOC_2018.pdf: 3661954 bytes, checksum: 45163caeab19a76f53c50788c263f240 (MD5) Previous issue date: 2018Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto de Ciência e Tecnologia em Biomodelos. Serviço de Controle da Qualidade Animal. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ. Brasil.MIGAL Technology Center. Virology and Vaccine Development Laboratory. Israel.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia Comparada e Ambiental. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ. Brasil.Group A rotaviruses (RVA) are one of the most common causes of severe acute gastroenteritis in infants worldwide. Rotaviruses spread from person to person, mainly by faecal⁻oral transmission. Almost all unvaccinated children may become infected with RVA in the first two years of life. The establishment of an experimental monkey model with RVA is important to evaluate new therapeutic approaches. In this study, we demonstrated viral shedding and viraemia in juvenile⁻adult Macaca fascicularis orally inoculated with Wa RVA prototype. Nine monkeys were inoculated orally: seven animals with human RVA and two control animals with saline solution. During the study, the monkeys were clinically monitored, and faeces and blood samples were tested for RVA infection. In general, the inoculated animals developed an oligosymptomatic infection pattern. The main clinical symptoms observed were diarrhoea in two monkeys for three days, associated with a reduction in plasmatic potassium content. Viral RNA was detected in seven faecal and five sera samples from inoculated animals, suggesting virus replication. Cynomolgus monkeys are susceptible hosts for human Wa RVA infection. When inoculated orally, they presented self-limited diarrhoea associated with presence of RVA infectious particles in faeces. Thus, cynomolgus monkeys may be useful as animal models to evaluate the efficacy of new antiviral approaches

Hepatitis E seroprevalence and associated factors in rural settlers in Central Brazil

<div><p>Abstract INTRODUCTION: Prevalence of hepatitis E virus (HEV) infection and associated factors were investigated in rural settlements in Central Brazil. METHODS: A total of 464 settlers were interviewed, and serum samples were tested for anti-HEV IgG/IgM. Positive samples were tested for HEV RNA. RESULTS: Sixteen participants (3.4%; 95% CI 2.0-5.7) were positive for anti-HEV IgG. None was positive for anti-HEV IgM. HEV RNA was not detected. Dwelling in a rural settlement for >5 years was associated with HEV seropositivity. CONCLUSIONS: The results revealed the absence of acute infection and a low prevalence of previous exposure to HEV.</p></div