RESUMO
Fourteen hepatitis C virus (HCV) chronically infected patients were submitted to routine liver biopsy for histological evaluation. Liver samples were assayed to HCV-RNA by in situ hybridization, using digoxigenin labeled probe. HCV genotypes were found to be predominantly type 1 (71.4%), followed by genotype 3 (21.4%), and genotype 2 (7.2%). Alanine-aminotransferase levels were raised in 10 patients. The histopathological scores were minimal (21.4%), mild (57.2%), and moderate (21.4%). Viral RNA was detected in liver cells from nine patients (64.3%). ISH method provides localization and poor confirmation of HCV RNA in the liver tissue of HCV chronic patients.
Assuntos
Hepacivirus/genética , Hepatite C Crônica/virologia , Hibridização In Situ/métodos , Fígado/virologia , RNA Viral/isolamento & purificação , Adulto , Idoso , Alanina Transaminase/sangue , Biópsia , Digoxigenina , Feminino , Formaldeído , Genótipo , Hepacivirus/isolamento & purificação , Hepatite C Crônica/patologia , Humanos , Fígado/patologia , Masculino , Pessoa de Meia-Idade , Inclusão em Parafina , Polimorfismo de Fragmento de Restrição , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Índice de Gravidade de DoençaRESUMO
Fourteen hepatitis C virus (HCV) chronically infected patients were submitted to routine liver biopsy for histological evaluation. Liver samples were assayed to HCV-RNA by in situ hybridization, using digoxigenin labeled probe. HCV genotypes were found to be predominantly type 1 (71.4 percent), followed by genotype 3 (21.4 percent), and genotype 2 (7.2 percent). Alanine-aminotransferase levels were raised in 10 patients. The histopathological scores were minimal (21.4 percent), mild (57.2 percent), and moderate (21.4 percent). Viral RNA was detected in liver cells from nine patients (64.3 percent). ISH method provides localization and poor confirmation of HCV RNA in the liver tissue of HCV chronic patients.
Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Hepacivirus/genética , Hepatite C Crônica/virologia , Hibridização In Situ/métodos , Fígado/virologia , RNA Viral/isolamento & purificação , Alanina Transaminase/sangue , Biópsia , Digoxigenina , Formaldeído , Genótipo , Hepacivirus/isolamento & purificação , Hepatite C Crônica/patologia , Fígado/patologia , Inclusão em Parafina , Polimorfismo de Fragmento de Restrição , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA Viral/genética , Índice de Gravidade de DoençaRESUMO
Common marmosets (Callithrixjacchus) were orally inoculated with a Brazilian strain (HAF-203) of hepatitis A virus (HAy). Three monkeys were euthanized at postinoculation hours 6, 12 and 24 to investigate the early events of HAV infection. Following others three inoculated and one control marmosets remained throughout the 46 day to evaluation of viral excretion. Different samples were collected to detect sequential presence of HAV RNA by nested reverse transcription-polymerase chain reaction (RT-PCR) in liver, saliva, bile and stools at 6 hours to 461h days postinoculation. Liver tissues were examined by immunofluorescence assay in a confocal laser-scanning microscope for the presence of HAV antigen. HAV RNA was detected in saliva during the course of the study, in bile from 24 hours to 46 days. in stools from 7 to 46 days and liver at 12 hours postinfection. In immunofluorescence of liver stained preparations, viral antigen was present at six hours after inoculation throughout the remainder of the 46-day study. The animals developed histological and biochemical acute hepatitis after second week postinoculation. Spleen, duodenum, and mesenteric lymph nodes specimens were negative for HAV antigens. This study supports the possibility that in Callithrixjacchus orally inoculated with hepatitis A virus the saliva route may be additional way of viral elimination. The viral replication in the liver was responsible for biliary HAV presence and latter HAV detection in fecal samples.
Assuntos
Antígenos Virais/análise , Callithrix , Vírus da Hepatite A/imunologia , Hepatite A/imunologia , Doenças dos Macacos/imunologia , Replicação Viral/imunologia , Animais , Modelos Animais de Doenças , Hepatite A/patologia , Hepatite A/transmissão , Antígenos da Hepatite A , Vírus da Hepatite A/crescimento & desenvolvimento , Vírus da Hepatite A/isolamento & purificação , Fígado/imunologia , Fígado/patologia , Fígado/virologia , Doenças dos Macacos/transmissão , RNA Viral/análise , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Callithrix jacchus is considered a reliable animal model for hepatitis A virus (HAV) infection. All three HAV orally inoculated marmosets developed hepatitis - the infection was monitored by continuous virus shedding, high levels of serum enzyme alanine aminotransferase, specific antibody and seroconversion 3-6 weeks after HAV inoculation. HAV antigen was detected in liver by immunofluorescence 4 days post inoculation (PI) and onwards. To gain insight into the biological role of inducible nitric oxide synthase (iNOS) during immune-related acute liver injury the enzyme was searched in frozen biopsies: immunofluorescent labeling was found in the cytoplasm of liver cells mainly Kupffer's cells and spleen macrophages (CD68+) starting 11 days PI with maximum intensity on the fifth to sixth week PI. Necroinflammatory liver lesions characteristic of viral hepatitis were also observed at 10 days PI with maximum severity at 4 to 6 weeks PI. Furthermore, T lymphocytes (CD2+) were raised at this time point. No difference was evident in the frequency of B lymphocytes (CD20+). Therefore, iNOS expression preceded necroinflammatory liver lesion and maximal immunofluorescence reaction was coincident with tissue injury, supporting the hypothesis that NO contributes to hepatic cytotoxic mechanism but also to virus clearance. The concomitant rise in T-lymphocyte population may suggest a role for these cells in this and/or other independent HAV-induced pathological changes.
Assuntos
Hepatite A/enzimologia , Hepatovirus , Fígado/patologia , Óxido Nítrico Sintase/biossíntese , Linfócitos T/imunologia , Animais , Callithrix , Modelos Animais de Doenças , Indução Enzimática , Imunofluorescência , Hepatite A/patologia , Imunofenotipagem , Fígado/enzimologia , Fígado/virologia , Necrose , Óxido Nítrico Sintase Tipo II , Baço/virologia , Linfócitos T/virologiaRESUMO
Hepatitis B infection and disease are highly endemic in South America. Prevalences of positivity are particularly high in Amazonia, and among Amerindian peoples in particular. This paper reports the results of a seroepidemiological survey for hepatitis B virus (HBV) carried out among four Amerindian populations from the Brazilian Amazon region: Gavião, Surui, Zoro and Navate. Rates of positivity to HBV serological markers (HBsAg, anti-HBs and or anti-HBc) are very high for the four groups, ranging from 62.8 to 95.7%. It is argued that the high rates of positivity in the Amerindian groups dealt with in this study, as well as for other Amazonian populations, are related to a complex of cultural practices which enhance the likelihood of HBV transmission (bloodletting, scarification, tattooing and orally processed food, among others). The authors suggest that, due to unique patterns of interaction between sociocultural and environmental factors. HBV infection assumes a specific profile in native Amazonian societies.
Assuntos
Cultura , Hepatite B/epidemiologia , Indígenas Sul-Americanos , Adolescente , Adulto , Distribuição por Idade , Brasil/epidemiologia , Distribuição de Qui-Quadrado , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Prevalência , Estudos Soroepidemiológicos , Distribuição por Sexo , Comportamento SocialRESUMO
Samples of serum, feces and liver tissue and organs of six cotton-eared marmosets Callithrix jacchus infected intravenously with two different strains of hepatitis A virus (HAV), were studied by conventional histologic techniques, by serological techniques and by immunocytochemical methods, such as immunofluorescence (IF) and peroxidase-antibody techniques. Hepatitis A antigen (HAAg) was detectable in daily collected stools, in liver biopsy obtained sequentially, and in organs collected at necropsy. Two marmosets also developed antibodies to HAV. By contrast, serum transaminases were not altered and there were histological hepatic lesions consistent with acute viral hepatitis in all inoculated animals. The data obtained, demonstrate that these primates are susceptible to human HAV and may be a useful animal model for the study of infection by this virus.
Assuntos
Callithrix , Hepatite A/veterinária , Animais , Antígenos Virais/análise , Fezes/microbiologia , Hepatite A/metabolismo , Hepatite A/patologia , Anticorpos Anti-Hepatite/análise , Hepatovirus/isolamento & purificação , Técnicas ImunoenzimáticasRESUMO
A previous seroepidemiological study in the rural zone of Vargem Alta (ES) SouthEast of Brazil, showed a prevalence of up to 9% of hepatitis B surface antigen (HBsAg) in some areas. One hundred susceptible children aging 1 to 5 years old were selected and immunized with a recombinant DNA hepatitis B vaccine (Smith-Kline 20 mcg) using the 0-1-6 months vaccination schedule. Blood samples were collected at the time of the first vaccine dose (month 0) in order to confirm susceptible individuals and 1,3,6 and 8 months after the first dose, to evaluate the antibody response. Our results showed that two and five months after the second dose, 79% and 88% of children seroconverted respectively, reaching 97% after the third dose. The levels of anti-HBs were calculated in milli International Units/ml (mIU/ml) and demonstrated the markedly increase of protective levels of antibodies after the third dose. These data showed a good immunogenicity of the DNA recombinant hepatitis B vaccine when administered in children of endemic areas.