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1.
J Mol Diagn ; 7(4): 535-43, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16237224

RESUMO

Pegylated alpha-interferon plus ribavirin is the current therapy for chronic hepatitis C virus (HCV) infection. Serum HCV-RNA concentration before treatment has been identified as an independent predictive factor of response. We have compared the percentage of HCV-infected hepatocytes with the concentration of serum HCV-RNA in baseline samples as predictors of response. We included 97 patients with chronic HCV infection (genotype 1), treated with pegylated-interferon-alpha2b plus ribavirin. Of these 97, 38 (39%) were sustained responders and 59 (61%) were not. Statistical differences between responders and nonresponders were found regarding the percentage of infected hepatocytes (6.83+/-4.50% versus 13.44+/-10.05%; P=0.00003) but not in serum HCV-RNA concentration [1.71+/-2.70 (x10(6) IU/L) versus 1.32+/-1.86 (x10(6) IU/L); P=0.40694]. Other factors associated with response were age, gamma-glutamyl transpeptidase level, and absence of previous therapy. Logistic regression demonstrated that percentage of infected hepatocytes (odds ratio, 1.160; 95% confidence interval, 1.065-1.264) and previous therapy (odds ratio, 0.294; 95% confidence interval, 0.109-0.795) were significant predictive factors for response. Therefore, the percentage of infected hepatocytes in liver biopsy before treatment is a better predictive factor of sustained response to 48 weeks of therapy with pegylated alpha-interferon plus ribavirin than serum HCV-RNA concentration in baseline serum sample.


Assuntos
Hepacivirus/isolamento & purificação , Hepatite C/diagnóstico , Hepatite C/tratamento farmacológico , Hepatócitos/virologia , Viremia/virologia , Adulto , Biópsia , Feminino , Hepacivirus/efeitos dos fármacos , Hepacivirus/fisiologia , Hepatite C/virologia , Humanos , Hibridização In Situ , Masculino , Pessoa de Meia-Idade , Razão de Chances , Prognóstico , RNA Viral/genética , Curva ROC , Reprodutibilidade dos Testes , Resultado do Tratamento , Viremia/sangue
2.
J Med Virol ; 73(2): 177-86, 2004 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15122790

RESUMO

Hepatitis B virus (HBV) DNA may persist in the liver in the absence of serum HBV-DNA after a self-limited acute hepatitis B. This may also occur in patients with chronic hepatitis C virus (HCV) infection but its prevalence and its impact on liver histology is unknown. HBV-DNA was tested by polymerase chain reaction (PCR) and by in situ hybridisation in liver biopsies from 98 patients with chronic hepatitis C who were hepatitis B surface antigen negative and serum HBV-DNA negative by PCR. HBV-DNA resulted positive in the liver of 37/98 (37.7%) patients without serum HBV-DNA. To test whether these patients had serum HBV-DNA levels under the detection limit of the PCR assay used in this study (50 copies/ml), PCR products in which HBV-DNA was undetectable after visualization of agarose gels were analysed by dot-blot hybridisation. With this method, HBV-DNA was positive in serum of 12/37 patients with liver HBV-DNA. Thus, 25/98 (25.5%) patients have HBV-DNA detectable only in liver. This was confirmed by in situ hybridisation, the percentage of infected hepatocytes ranging from 0.1% to 12%. In patients in whom the HCV infection was shorter than 20 years, HBV infected patients had higher (P = 0.01) fibrosis score (1.64 +/- 1.21) than HBV negative cases (0.53 +/- 0.66). In conclusion, a significant proportion of patients with chronic HCV infection have HBV-DNA in the liver in the absence of viral DNA in serum. The impact of this finding on liver histology deserves further research.


Assuntos
DNA Viral/análise , DNA Viral/sangue , Vírus da Hepatite B/isolamento & purificação , Hepatite B/complicações , Hepatite B/virologia , Hepatite C Crônica/complicações , Fígado/virologia , Adulto , Sequência de Bases , Biópsia , DNA Viral/química , Feminino , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/genética , Hepatócitos/virologia , Humanos , Hibridização In Situ , Fígado/patologia , Cirrose Hepática/patologia , Cirrose Hepática/virologia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Estudos Prospectivos , Alinhamento de Sequência , Análise de Sequência de DNA
3.
J Infect Dis ; 189(1): 7-14, 2004 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-14702147

RESUMO

BACKGROUND: There are patients in whom the etiology of long-standing abnormal results of liver-function tests is unknown (ALF-EU) after exclusion of all known causes of liver diseases. We analyzed the presence of hepatitis C virus (HCV) RNA in liver-biopsy specimens from 100 patients who were negative for anti-HCV antibodies and for serum HCV RNA and who had ALF-EU. METHODS: HCV RNA status was tested by reverse-transcription polymerase chain reaction (RT-PCR) and by in situ hybridization, in liver and peripheral-blood mononuclear cells (PBMCs). RESULTS: HCV RNA was detected in liver-biopsy specimens from 57 of 100 patients negative for anti-HCV antibodies and for serum HCV RNA (i.e., who had occult HCV infection). HCV RNA of negative polarity was found in the liver of 48 (84.2%) of these 57 patients with occult HCV infection. Nucleotide-sequence analysis confirmed the specificity of detection of HCV RNA and that patients were infected with the HCV 1b genotype. Of these 57 patients with intrahepatic HCV RNA, 40 (70%) had viral RNA in their PBMCs. With regard to liver histology, patients with occult HCV infection were more likely to have necroinflammatory activity (P=.017) and fibrosis (P=.022) than were patients without intrahepatic HCV RNA. CONCLUSIONS: Patients with ALF-EU may have intrahepatic HCV RNA in the absence of anti-HCV antibodies and of serum HCV RNA.


Assuntos
Hepacivirus/isolamento & purificação , Hepatite C/diagnóstico , Hepatopatias/etiologia , Fígado/enzimologia , RNA Viral/análise , Adulto , Idoso , Alanina Transaminase/sangue , Biópsia , Estudos de Coortes , Feminino , Genótipo , Glutamil Aminopeptidase/sangue , Hepacivirus/genética , Hepacivirus/patogenicidade , Hepatite C/sangue , Hepatite C/complicações , Anticorpos Anti-Hepatite C/sangue , Humanos , Hibridização In Situ , Leucócitos Mononucleares/virologia , Fígado/virologia , Hepatopatias/enzimologia , Testes de Função Hepática , Masculino , Pessoa de Meia-Idade , RNA Viral/sangue , Reação em Cadeia da Polimerase Via Transcriptase Reversa , gama-Glutamiltransferase/sangue
4.
J Med Virol ; 70(4): 571-80, 2003 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12794719

RESUMO

Although occult hepatitis B virus (HBV) infection (HBV-DNA in serum in the absence of hepatitis B surface antigen [HBsAg]) is common in chronic hepatitis C, its characteristics are not well known. In this work, the presence of HBV-DNA (by polymerase chain reaction; PCR) and its distribution (by in situ hybridization) in liver biopsies and peripheral blood mononuclear cells (PBMCs) from 32 patients with chronic hepatitis C and occult HBV infection and in 20 HBsAg chronic carriers were determined. The results showed that serum HBV-DNA levels were statistically lower (P = 0.001) in patients with occult HBV infection than in HBsAg chronic carriers. The HBV infection pattern in liver cells was identical between patients with occult HBV infection and those with chronic hepatitis B. However, the mean percentage of HBV-infected hepatocytes was significantly lower (P = 0.001) in patients with occult HBV infection (5 +/- 4.44%) than in HBsAg chronic carriers (17.99 +/- 11.58%). All patients with chronic hepatitis B have HBV-DNA in their PBMCs while this occurred in 50% of the cases with occult HBV infection. In conclusion, patients with occult HBV infection have a low number of HBV-infected hepatocytes and this fact could explain the lack of HBsAg detection and low viremia levels found in these cases.


Assuntos
DNA Viral/sangue , Vírus da Hepatite B/isolamento & purificação , Hepatite B/complicações , Hepatite C Crônica/complicações , Fígado/virologia , Viremia/virologia , Adulto , Biópsia , DNA Viral/análise , Feminino , Hepatite B/virologia , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/genética , Hepatócitos/virologia , Humanos , Hibridização In Situ , Fígado/citologia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase
5.
J Med Virol ; 68(4): 529-36, 2002 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-12376961

RESUMO

Hepatitis C virus (HCV) replicates in salivary glands of chronic hepatitis C patients and is released into the saliva, suggesting that HCV may replicate in other exocrine glands. The presence of positive and negative HCV RNA strands was demonstrated by in situ hybridization, and of HCV core protein by immunohistochemistry, in sweat glands and keratinocytes in healthy skin biopsies from 15 patients with chronic hepatitis C and 10 anti-HCV negative patients with chronic liver disease. Positive and negative HCV RNA strands were detected in 9.6 +/- 5.2% and 4.2 +/- 3.8%, respectively, of the epithelial cells of eccrine sweat glands. Core protein was detected in 6.0 +/- 3.93% of these cells. HCV RNA resistant to RNase digestion (encapsidated HCV RNA) was detected in 10/10 sweat samples from HCV-infected patients. Positive and negative HCV RNA strands were detected in 6.7 +/- 2.97% and 3.0 +/- 3.08% of the keratinocytes, respectively. HCV core protein was found in 4.5 +/- 2.76% of these cells. No HCV RNA or HCV core protein was detected in the skin biopsies from the 10 anti-HCV negative patients. In conclusion, HCV replicates in eccrine sweat glands cells and keratinocytes in healthy skin and is released into the sweat.


Assuntos
Hepacivirus/fisiologia , Hepatite C Crônica/virologia , Queratinócitos/virologia , Glândulas Sudoríparas/virologia , Replicação Viral , Adulto , Feminino , Hepacivirus/isolamento & purificação , Humanos , Imuno-Histoquímica , Hibridização In Situ , Masculino , Pessoa de Meia-Idade , RNA Viral/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas do Core Viral/análise
6.
J Invest Dermatol ; 119(4): 798-803, 2002 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-12406323

RESUMO

Cutaneous lichen planus has been associated in patients with chronic hepatitis C virus infection. It is still unknown whether hepatitis C virus infects keratinocytes of lichen planus lesions. In this report we have analyzed the presence of genomic and anti-genomic hepatitis C virus RNA in skin biopsies from 26 patients with chronic hepatitis C and healthy skin and from 24 patients with cutaneous lichen planus (five with and 19 without hepatitis C virus infection) by in situ hybridization. Hepatitis C virus RNA was detected in the keratinocytes of 69% of the patients with healthy skin and chronic hepatitis C, in 100% of the patients with lichen planus and hepatitis C virus infection, and in none of lichen planus patients without hepatitis C virus infection. The percentage of keratinocytes showing genomic or anti-genomic hepatitis C virus RNA was statistically lower (p < 0.01 in all cases) in patients with healthy skin (mean +/- SD: 5.7 +/- 3.5% and 2.7 +/- 3.1% of keratinocytes with genomic or anti-genomic hepatitis C virus RNA, respectively) than in those with lichen planus lesions (31.7 +/- 7.9% and 18.8 +/- 7.4%, mean +/- SD) or the unaffected adjacent skin (24.8 +/- 6.9% and 14.3 +/- 3.8%, mean +/- SD). In conclusion, we have demonstrated that hepatitis C virus infects keratinocytes from patients with lichen planus and hepatitis C virus infection.


Assuntos
Hepatite C Crônica/virologia , Queratinócitos/virologia , Líquen Plano/virologia , RNA Viral/análise , Proteínas do Core Viral/análise , Adulto , Idoso , Feminino , Hepacivirus/química , Hepacivirus/genética , Humanos , Imuno-Histoquímica , Hibridização In Situ , Masculino , Pessoa de Meia-Idade , RNA Viral/sangue , Proteínas S100/análise
7.
Virology ; 301(1): 121-9, 2002 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-12359452

RESUMO

TT virus (TTV) is an unenveloped, single-stranded, circular-DNA virus which resembles members of the Circoviridae, that is commonly found in humans and which lacks pathological consequences for the infected host. TTV replication has been demonstrated in bone marrow cells but not in peripheral blood mononuclear cells (PBMC), suggesting that hematopoietic cells must be activated to support TTV replication. To test this hypothesis, PBMC from two naturally TTV-infected individuals and from two healthy TTV-DNA negative donors infected in vitro with a TTV-DNA-positive serum were cultured in the presence (stimulated) or absence (unstimulated) of phytohemagglutinin, lipopolysaccharide, and interleukin-2. TTV-DNA was detected in both stimulated and unstimulated PBMC. However, TTV-DNA replicative intermediates and mRNA were detected only in stimulated PBMC. Furthermore, TTV-DNA and mRNA were detected in PBMC from two TTV negative donors reinfected with supernatants from TTV-infected stimulated cells but not when using culture supernatants from unstimulated cells. These results demonstrate that TTV replicates in PBMC only when stimulated.


Assuntos
Leucócitos Mononucleares/virologia , Torque teno virus/fisiologia , Replicação Viral , Células Cultivadas , DNA Viral/análise , Humanos , Hibridização in Situ Fluorescente , RNA Viral/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Torque teno virus/genética
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