Evidences of Hepatitis C Virus Replication in Hepatocytes and Peripheral Blood Monocuclear Cells from Patients Negative for Viral RNA in Serum

Abstract

Abstract:In this study, we examined the expression of hepatitis C virus (HCV) components in hepatocytes and peripheral blood mononuclear cells (PBMC) from patients positive for anti-HCV antibodies and negative for serum HCV-RNA. The samples obtained from 25 randomly selected patients (13 of 25 patients showed no histological evidences of chronic hepatitis and had normal serum ALAT and GGTP levels) were studied by in situ Hybridization and Immunofluorescence assays. The findings show that HCV-RNA of both positive and negative polarity was carried in the hepatocytes of more than 80% of cases. The proportion of cells expressing the negative strand (mean ± SD, 10.25% ± 5.56%) was lower than those expressing positive strand (mean ± SD, 19.88% ± 9.19%) (p=0.0076; Student's t test). In addition, reaction products suggestive of HCV core, E1 and E2 antigens within hepatocytes were also observed. Both hybridization and immunofluorescence signals were localized in the cytoplasm suggesting that this is the place of active HCV replication. On the other hand, the HCV-RNA of positive polarity was detected in PBMC from 16 out of 17 samples analyzed (94%) while the HCV-RNA of negative polarity was detected in 82% of samples investigated. Positive hybridization signals were localized in the cytoplasm of PBMC. Interestingly, 12 out of 13 patients with clinical and histological resolution of hepatitis, contain HCV-RNA in either liver or PBMC. These results provide further evidences that the intermediate replicative form of the HCV genome can persist in hepatocytes and PBMC after apparently complete resolution of chronic hepatitis C.

Occult hepatitis B virus and hepatitis C virus infections

“…In this population, known as 'healthy' HCV carriers, HCV infection and replication has been detected in liver and PBMC of nearly 90% of the subjects [88][89][90]. (B) in patients with chronic hepatitis C who have 'cleared' HCV infection, either spontaneously or after a successful antiviral treatment [91][92][93].…”

Evidences of Apoptosis in Hepatitis C Virus-Infected Hepatocytes and Peripheral Blood Monocuclear Cells in the Absence of Liver Injury

“…These 21 patients were selected based upon they showed negative detection of serum HCV-RNA by RT-PCR (Amplicor HCV Amplification Kit 2.0, Roche Diagnostic Systems,Inc) on the same day of the liver biopsy. In addition, all patients showed positive detection of intrahepatic HCV-RNA of both positive and negative polarity [11] . None was seropositive for markers of hepatitis B virus (HBV), hepatitis A virus and human immunodeficiency virus by enzyme immunoassay (Tecnosuma International, Havana, Cuba).…”

“…In addition, PBMC samples from 14 of these 21 selected patients taken on the same day of the liver biopsy were analyzed. They showed positive detection of HCV-RNA of positive and negative polarity [11] . A total 11 out of 21 patients showed no histological evidences of chronic hepatitis and had normal serum alanine aminotransferase (ALAT) or gamma-glutamyl transpeptidase (GGTP) levels for at least six month before the biopsy was performed.…”

“…The total number of 21 patients used in this study have been recently described [11] . They were serologically positive to third-generation HCV enzyme immunoassays (Tecnosuma International, Havana, Cuba).…”

“…In addition, it has been shown that HCV-RNA can persist at very low levels in the serum, PBMC and liver for many years after apparently complete spontaneous or antiviral therapy-induced resolution of chronic hepatitis C [9,10] . Recently, it has been shown the presence of HCV-RNA of both positive and negative polarity in hepatocytes and PBMC from patients positive for anti-HCV antibodies and negative for serum HCV-RNA [11] . In the present work, the presence of apoptosis in HCV-infected liver and PBMC from 21 of these patients was investigated..…”