RESUMO
BACKGROUND: Nonalcoholic steatohepatitis (NASH) is a chronic inflammatory liver disease influenced by risk factors for the metabolic syndrome. In adult patients, NASH is associated with an altered phenotype and functionality of peripheral immune cells, the recruitment of leukocytes and intrahepatic activation, and an exacerbated production of reactive oxygen species (ROS) and cytokines. It remains unclear if the previously described differences between pediatric and adult nonalcoholic fatty liver diseases also reflect differences in their pathogenesis. AIMS: We aimed to investigate the phenotype and functionality of circulating immune cells and the potential contribution of liver infiltrating leukocytes to the immunological imbalance in pediatric NASH. RESULTS: By a real-time PCR-based analysis of cytokines and immunohistochemical staining of liver biopsies, we demonstrated that the hepatic microenvironment is dominated by interferon-gamma (IFN-γ) but not interleukin-4 and is infiltrated by a higher number of CD8(+) cells in pediatric NASH. The number of infiltrating neutrophils positively correlated with ROS generation by peripheral polymorphonuclear cells. By a flow cytometric analysis of peripheral blood lymphocytes, a distinctive increase in CD8(+) CD45RO and CD8(+) CD45RA subpopulations and an increased production of IFN-γ by CD4(+) and CD8(+) cells were shown. The production of ROS following PMA stimulation was augmented in circulating neutrophils but not in monocytes. CONCLUSION: In sum, the distinctive phenotype and functionality of infiltrating and circulating cells suggest that the role of innate cells is coupled to a Th1-polarized immune response in pediatric NASH.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Fígado Gorduroso/imunologia , Adolescente , Linfócitos T CD8-Positivos/imunologia , Criança , Fígado Gorduroso/patologia , Feminino , Humanos , Interferon gama/genética , Interferon gama/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Leucócitos Mononucleares/imunologia , Masculino , Neutrófilos/imunologia , Hepatopatia Gordurosa não Alcoólica , RNA Mensageiro/imunologia , Espécies Reativas de Oxigênio/imunologiaRESUMO
INTRODUCTION: Neonatal cholestasis due to endocrine diseases is infrequent and poorly reco-gnized. Referral to the pediatric endocrinologist is delayed. OBJECTIVE: We characterized cholestasis in infants with congenital pituitary hormone deficiencies (CPHD), and its resolution after hormone replacement therapy (HRT). SUBJECTS AND METHODS: Sixteen patients (12 males) were included; eleven with CPHD, and five with isolated central hypocortisolism. RESULTS: Onset of cholestasis occurred at a median age of 18 days of life (range 2-120). Ten and nine patients had elevated transaminases and γGT, respectively. Referral to the endocrinologist occurred at 32 days (range 1 - 72). Remission of cholestasis occurred at a median age of 65 days, whereas liver enzymes occurred at 90 days. In our cohort isolated, hypocortisolism was a transient disorder. CONCLUSION: Cholestasis due to hormonal deficiencies completely resolved upon introduction of HRT. Isolated hypocortisolism may be a transient cause of cholestasis that needs to be re-evaluated after remission of cholestasis.
INTRODUÇÃO: A colestase neonatal causada por doenças endócrinas é pouco frequente e reconhecida. Existe um atraso no encaminhamento dos pacientes a um endocrinologista pediátrico. OBJETIVO: Caracterizamos a colestase em recém-nascidos com deficiências congênitas de hormônio hipofisário (DCHH) e sua resolução após a terapia de reposição hormonal (TRH). SUJEITOS E MÉTODOS: Dezesseis pacientes (12 do sexo masculino) foram incluídos; sete com DCHH, e cinco com hipocortisolismo central isolado. RESULTADOS: O início da colestase ocorreu aos 18 dias de vida (variação 2-120). Dez e nove pacientes apresentaram elevação das transaminases e γGT, respectivamente. A consulta com um endocrinologista aconteceu aos 32 dias (variação 1-72). A remissão da colestase ocorreu em uma idade mediana de 65 dias, enquanto a remissão das enzimas hepáticas aconteceu aos 90 dias. Na coorte isolada, o hipocortisolismo foi uma desordem transitória. CONCLUSÃO: A colestase causada por deficiências hormonais foi completamente resolvida após a introdução da TRH. O hipocortisolismo pode ser uma causa transitória da colestase e precisa ser reavaliado após a remissão da colestase.
Assuntos
Feminino , Humanos , Lactente , Masculino , Insuficiência Adrenal/etiologia , Colestase/etiologia , Hidrocortisona/uso terapêutico , Hipopituitarismo/congênito , Hepatopatias/etiologia , Tiroxina/uso terapêutico , Idade de Início , Insuficiência Adrenal/fisiopatologia , Colestase/fisiopatologia , Seguimentos , Terapia de Reposição Hormonal/métodos , Hidrocortisona/deficiência , Hipopituitarismo/tratamento farmacológico , Hepatopatias/fisiopatologia , Hormônios Adeno-Hipofisários/deficiência , Indução de Remissão , Estudos Retrospectivos , Resultado do TratamentoRESUMO
INTRODUCTION: Neonatal cholestasis due to endocrine diseases is infrequent and poorly recognized. Referral to the pediatric endocrinologist is delayed. OBJECTIVE: We characterized cholestasis in infants with congenital pituitary hormone deficiencies (CPHD), and its resolution after hormone replacement therapy (HRT). SUBJECTS AND METHODS: Sixteen patients (12 males) were included; eleven with CPHD, and five with isolated central hypocortisolism. RESULTS: Onset of cholestasis occurred at a median age of 18 days of life (range 2-120). Ten and nine patients had elevated transaminases and γGT, respectively. Referral to the endocrinologist occurred at 32 days (range 1 - 72). Remission of cholestasis occurred at a median age of 65 days, whereas liver enzymes occurred at 90 days. In our cohort isolated, hypocortisolism was a transient disorder. CONCLUSION: Cholestasis due to hormonal deficiencies completely resolved upon introduction of HRT. Isolated hypocortisolism may be a transient cause of cholestasis that needs to be re-evaluated after remission of cholestasis.
Assuntos
Insuficiência Adrenal/etiologia , Colestase/etiologia , Hidrocortisona/uso terapêutico , Hipopituitarismo/congênito , Hepatopatias/etiologia , Tiroxina/uso terapêutico , Insuficiência Adrenal/fisiopatologia , Idade de Início , Colestase/fisiopatologia , Feminino , Seguimentos , Terapia de Reposição Hormonal/métodos , Humanos , Hidrocortisona/deficiência , Hipopituitarismo/tratamento farmacológico , Lactente , Hepatopatias/fisiopatologia , Masculino , Hormônios Adeno-Hipofisários/deficiência , Indução de Remissão , Estudos Retrospectivos , Resultado do TratamentoRESUMO
The immunopathogenesis of type I autoimmune hepatitis (AIH-I) might involve the deregulation of different cellular processes. Here, we investigated the liver expression of selected cytokines and genes of regulatory cell populations in children both at diagnosis and during biochemical remission following immunosuppressive treatment (AIH-Ir). We found a higher Vα24, IFN-γ, FoxP3, IL-27p28, IL-12p40 and IL-21 expression at diagnosis as well as a positive correlation between IL-21 and transaminase levels. Interestingly, only IFN-γ and FoxP3 were decreased in AIH-Ir. An "AIH-I phenotype" (high Vα24, IFN-γ and FoxP3 expression at diagnosis) was observed in only 5 out of 22 AIH-Ir patients but not in controls. These results indicate a local deregulation of the innate and adaptive immune responses with an increased transcriptional activity of immunoregulatory cells at diagnosis. In addition, IL-21 is highlighted as a mediator of liver injury. AIH-Ir is characterized by a partial reversal of the deregulated response.
Assuntos
Fatores de Transcrição Forkhead/metabolismo , Hepatite Autoimune/metabolismo , Interferon gama/metabolismo , Fígado/metabolismo , Receptores de Antígenos de Linfócitos T/metabolismo , Adolescente , Biomarcadores/metabolismo , Criança , Pré-Escolar , Feminino , Fatores de Transcrição Forkhead/genética , Regulação da Expressão Gênica , Hepatite Autoimune/imunologia , Humanos , Interferon gama/genética , Subunidade p40 da Interleucina-12/metabolismo , Interleucinas/metabolismo , Fígado/imunologia , Masculino , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , Receptores de Antígenos de Linfócitos T/genética , Transaminases/metabolismoRESUMO
Infection with hepatitis C virus (HCV) is a worldwide health problem with more than 170 million infected individuals. In children, since 1992 almost all infections occurred through vertical transmission from an infected mother to her newborn infant. Natural history of HCV infection in children is not yet well defined, most children are asymptomatic and may remain so for decades. Most infected individuals (60-80%), regardless of their age at infection, become chronically infected with HCV. Spontaneous resolution in children appears to be infrequent. Positive HCV antibody implicate that patient has been exposed to the virus (EIA test). To discriminate between active or resolved HCV viral infection it is necessary to perform HCV RNA (PCR). Liver biopsy assess degree of liver injury and exclude concurrent diseases. HCV chronic infection is slow progressive in childhood. Progression of fibrosis seems to be a function of infection duration. Treatment objective is clearance of HCVRNA. IFNa is recognized as the drug approved for hepatitis C treatment in pediatric population. Combination therapy with IFNa or pegylated IFNa plus ribavirin is recently approved by US FDA and EMEA and clinical trials are in progress.
Assuntos
Antivirais/uso terapêutico , Hepatite C Crônica/tratamento farmacológico , Infecções Assintomáticas , Biópsia , Criança , Pré-Escolar , Quimioterapia Combinada , Genótipo , Hepacivirus/genética , Hepacivirus/imunologia , Anticorpos Anti-Hepatite C/sangue , Hepatite C Crônica/diagnóstico , Hepatite C Crônica/epidemiologia , Hepatite C Crônica/transmissão , Humanos , Lactente , Recém-Nascido , Transmissão Vertical de Doenças Infecciosas , Interferon-alfa/uso terapêutico , RNA Viral/sangue , Ribavirina/uso terapêutico , Fatores de Risco , Resultado do Tratamento , Carga ViralRESUMO
To investigate the immunopathogenic mechanisms of type I autoimmune hepatitis in children, we analyzed by quantitative or semiquantitative reverse transcription-polymerase chain reaction the expression of cytokines interferon (IFN)-gamma, interleukin (IL)-12p40, IL-18, IL-4, IL-10, and IL-12R beta 2. In addition, liver and peripheral blood was collected to investigate the expression of the natural killer T (NKT) cell marker V alpha 24. The presence of NKT cells in hepatic lesions were also identified by immunohistochemistry. The analysis was performed on liver biopsies from 25 children with type I autoimmune hepatitis. As disease controls, we included six children with hepatitis C virus-related chronic hepatitis and nine control livers. The expression of IFN-gamma and IL-12p40 was not detected in controls but was clearly upregulated in pathologic biopsies. In addition, these samples showed an increased expression of IL-18 (p = 0.0003), IL-4 (p = 0.0055), and IL-12R beta 2 (p = 0.007). Western blot analysis confirmed the expression of IL-12p40 and IL-18. However, for IL-18, we detected only the immature biologically inactive polypeptide. The V alpha 24 transcripts were found increased in the liver (p = 0.0007) where V alpha 24(+) cells were also localized, but decreased in peripheral blood mononuclear cells (p = 0.041). In addition to a type I immune response, NKT cells might play a substantial role in the pathogenesis of type I autoimmune hepatitis in children.
Assuntos
Citocinas/genética , Expressão Gênica , Hepatite Autoimune/patologia , Interleucina-4/genética , Células Th1/imunologia , Adolescente , Autoanticorpos/sangue , Biópsia por Agulha , Análise Química do Sangue , Western Blotting , Criança , Citocinas/imunologia , Citocinas/metabolismo , Feminino , Hepatite Autoimune/genética , Hepatite Autoimune/imunologia , Humanos , Imuno-Histoquímica , Interferon gama/genética , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-12/genética , Interleucina-12/metabolismo , Subunidade p40 da Interleucina-12 , Interleucina-18/genética , Interleucina-18/metabolismo , Interleucina-4/imunologia , Interleucina-4/metabolismo , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Células Matadoras Naturais/patologia , Leucócitos Mononucleares/química , Fígado/imunologia , Fígado/metabolismo , Fígado/patologia , Masculino , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Receptores de Antígenos de Linfócitos T alfa-beta/análise , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Receptores de Interleucina/genética , Receptores de Interleucina/metabolismo , Receptores de Interleucina-12 , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismo , Células Th1/metabolismoAssuntos
Colestase , Atresia Biliar/terapia , Criança , Fenômenos Fisiológicos da Nutrição Infantil , Colestase/induzido quimicamente , Colestase/diagnóstico , Colestase/etiologia , Colestase/terapia , Colestase Intra-Hepática/diagnóstico , Colestase Intra-Hepática/etiologia , Colestase Intra-Hepática/terapia , Gastroenterologia , Humanos , Cooperação Internacional , Hepatopatias/diagnóstico , Hepatopatias/etiologia , Hepatopatias/terapia , Transplante de Fígado , Guias de Prática Clínica como Assunto , Pesquisa , Sociedades MédicasRESUMO
Hepatitis C virus (HCV) infection in children was assessed by RT-nested PCR of the 5'untranslated region (5'UTR) of the viral genome combined with virus genotyping, performed by restriction fragment length polymorphism (RFLP). We analysed HCV infection in 64 children and in 9 HCV chronically infected mothers corresponding to 10 of them. Thirty two children were positive for serum HCV RNA as determined by RT-nested PCR. The viremia was analysed in consecutive samples of 25 children. Nine children (36 per cent) were always positive for HCV RNA, in 5 (20 per cent) a positive RT-nested PCR turned negative in subsequent samples, other 9 (36 per cent) showed alternating RT-nested PCR results and in 2 (8 per cent) the RT-nested PCR turned positive after an initial negative result. The HCV genotype distribution was studied in 27/32 children and in 9 mothers, and it was similar to that reported in the literature for adult and pediatric patients in our country. Genotype 1 was predominant in our population. HCV genotype did not change in the same patient during the time of this study. HCV genotype was the same in mother-infant pairs. We could not establish a correlation between HCV genotype and vertical transmission of HCV. This study will be helpful to further analyze HCV behavior during pediatric infection and the host's response in the initial stages of it.
Assuntos
Humanos , Masculino , Feminino , Lactente , Pré-Escolar , Criança , Adolescente , Adulto , Hepacivirus , Hepatite C , Seguimentos , Genótipo , Hepatite C , Transmissão Vertical de Doenças Infecciosas , Polimorfismo de Fragmento de Restrição , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA ViralRESUMO
Hepatitis C virus (HCV) infection in children was assessed by RT-nested PCR of the 5untranslated region (5UTR) of the viral genome combined with virus genotyping, performed by restriction fragment length polymorphism (RFLP). We analysed HCV infection in 64 children and in 9 HCV chronically infected mothers corresponding to 10 of them. Thirty two children were positive for serum HCV RNA as determined by RT-nested PCR. The viremia was analysed in consecutive samples of 25 children. Nine children (36 per cent) were always positive for HCV RNA, in 5 (20 per cent) a positive RT-nested PCR turned negative in subsequent samples, other 9 (36 per cent) showed alternating RT-nested PCR results and in 2 (8 per cent) the RT-nested PCR turned positive after an initial negative result. The HCV genotype distribution was studied in 27/32 children and in 9 mothers, and it was similar to that reported in the literature for adult and pediatric patients in our country. Genotype 1 was predominant in our population. HCV genotype did not change in the same patient during the time of this study. HCV genotype was the same in mother-infant pairs. We could not establish a correlation between HCV genotype and vertical transmission of HCV. This study will be helpful to further analyze HCV behavior during pediatric infection and the hosts response in the initial stages of it. (Au)
Assuntos
Humanos , Masculino , Feminino , Lactente , Pré-Escolar , Criança , Adolescente , Adulto , Hepatite C/genética , Hepacivirus/genética , Hepatite C/transmissão , Genótipo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Seguimentos , Transmissão Vertical de Doenças Infecciosas , RNA Viral/sangue , Polimorfismo de Fragmento de RestriçãoRESUMO
HCV genomic characterization was performed by nucleotide sequence analysis (n=50) combined with restriction fragment length polymorphism (RFLP) of the 5'UTR region in 82 isolates coresponding to different Argentine groups. Genotype 1 was detected in 70.7 percent of the samples (58 out of 82), genotype 2 in 21.9 percent (18 of 82) and genotypes 3 in the remaining 6 sera (7.3 percent). HCV ib subtype contributed with 35.3 percent to the whole population studied (29 of 82) and was detected in 6 out of 21 sporadic cases. Besides their epidemiological significance, these results should be taken into account when future vaccines are considered on the basis of geographical HCV genotypic prevalence.
Assuntos
Adulto , Pessoa de Meia-Idade , Pré-Escolar , Criança , Feminino , Humanos , Adolescente , Hepacivirus/genética , Hepatite C Crônica/sangue , Filogenia , Polimorfismo de Fragmento de Restrição , Argentina , Sequência de Bases , Genótipo , Hepatite D Crônica , Reação em Cadeia da Polimerase , Fatores de Risco , Análise de Sequência de RNARESUMO
HCV genomic characterization was performed by nucleotide sequence analysis (n=50) combined with restriction fragment length polymorphism (RFLP) of the 5UTR region in 82 isolates coresponding to different Argentine groups. Genotype 1 was detected in 70.7 percent of the samples (58 out of 82), genotype 2 in 21.9 percent (18 of 82) and genotypes 3 in the remaining 6 sera (7.3 percent). HCV ib subtype contributed with 35.3 percent to the whole population studied (29 of 82) and was detected in 6 out of 21 sporadic cases. Besides their epidemiological significance, these results should be taken into account when future vaccines are considered on the basis of geographical HCV genotypic prevalence. (AU)