Detection of replicative form of HCV RNA in peripheral blood leukocytes and its clinical significance

Abstract

Nested RT-PCR, done by using degenerated primer pair, was used to detect hepatitis C virus RNA (HCV RNA) in serum, plasma, liver and peripheral blood mononuclear cells (PBMC) of 30 patients with acute and chronic posttransfusion hepatitis C and 7 asymptomatic anti-HCV positive subjects. The results showed that the percentages of both the plus and minus strands of HCV RNA in PBMC of the patients with chronic hepatitis C was significantly higher than that with acute hepatitis C and asymptomatic anti-HCV positive subjects (P < 0. 05–0. 001). In 17 patients who were subjected to biopsy, the positive rate of the both strands of HCV RNA in PBMC of the patients with AH was lower than that of CAH (P <0. 05). In serum and plasma of all 37 cases, the minus strand of HCV RNA was not detected. Both plus and minus strands in liver of one patient with AH were positive, but the minus strand in PBMC negative. In 6 patients with CAH whose both strands in liver were positive, both strands in PBMC in 5 patients were also found. The present data confirmed that PBMC of the patients with hepatitis C were infected by HCV and the longer the infection time, the bigger the possibility of PBMC infection by HCV. The patients with active liver disease (CAH) had higher positive rate of minus strands of HCV RNA in PBMC. The results suggested that HCV may not only infect PBMC but also replicate in PBMC, and that the occurrence of minus strand of HCV RNA is associated with activity of liver disease.

Keywords

hepatitis C virus RNA / nested RT-PCR / peripheral blood mononuclear cells

References

[1] MillerR H, PurcellR H. Hepatitis C virus shares amino acid sequence similarity with pestiviruses and flaviviruses as well as members of two plant virus supergroups. Proc Natl Acad Sci USA, 1990, 87: 2057-2057

[2] FongT L, ShindoM, FeinstoneSM, et al.. Detection of replicative intermediates of hepatitis C viral RNA in liver and serum of patients with chronic hepatitis C. J Clin Invest, 1991, 88: 1058-1058

[3] RNA 1994, 74: 284.

[4] WangJ T, SheuT C, LinJ T, et al.. Detection of replicative form of hepatitis C virus RNA in peripheral blood mononuclear cells. J Infect Dis, 1992, 166: 1167-1167

[5] TakeharaT, HayashiN, MitaE, et al.. Detection of the minus strand of hepatitis C virus RNA by reverse transcription and polymerase chain reaction: implications for hepatitis C virus replication in infected tissue. Hepatology, 1992, 15: 387-387

[6] GilB, QianC, Riezu-BojJ I, et al.. Hepatic and extrahepatic HCV RNA strands in chronic hepatitis C: different patterns of response to interferon treatment. Hepatology, 1993, 18: 1050-1050

[7] DNA. PCR 1992. 62–68.

[8] SambrookJ, FritschE F, ManiatisT. Molecular Cloning: A laboratory manual. 2nd, 1989New YorkCold Spring Harbor11.5-11.16

[9] MullerH M, PfaffE, GoeserT, et al.. Peripheral blood leukocytes serve as a possible extrahepatic site for hepatitis C virus replication. J Gen Virol, 1993, 74: 669-669