HCV-RNA ASSAY IN PERIPHERAL BLOOD MONONUCLEAR CELLS IN RELATION TO IFN THERAPY.

Okuda M; First Department of Internal Medicine, Yamaguchi University School of Medicine, Ube, Japan. Gastroenterol Jpn, 1993 Aug, 28:4, 535-40

Hepatitis C virus RNA (HCV-RNA) was serially assayed in the serum and peripheral blood mononuclear cells (PBMC) of 10 patients with chronic hepatitis C who underwent interferon (IFN) therapy, and whether detection of HCV-RNA from PBMC serves as an index of the response of chronic hepatitis C to IFN therapy was evaluated. HCV-RNA was assayed by reversed transcription and polymerase chain reaction using the 5'-noncoding region as a primer. IFN therapy was effective in 3 patients and ineffective in the other 7 patients. HCV-RNA disappeared from the serum during and immediately after the IFN therapy in all 3 patients in whom the therapy was effective and in 3 of 7 patients in whom the therapy was ineffective. HCV-RNA disappeared from PBMC in all 3 patients in whom the therapy was effective, but PBMC HCV-RNA remained positive in 6 of the 7 patients in whom the therapy was ineffective, and the serum HCV-RNA became positive again in 5 of these 6 patients after 6 months. The disappearance of HCV-RNA from PBMC was associated with long-term stabilization of the serum alanine aminotransferase value, so HCV-RNA assay in PBMC is considered to be useful as a prognostic marker of chronic hepatitis C after IFN therapy.

GENOME DETECTION IN LIVER AND PERIPHERAL BLOOD MONONUCLEAR CELLS: PREDICTOR FACTORS OF SUSTAINED RESPONSE IN PATIENTS WITH CHRONIC HEPATITIS C.

Berenguer M; Olaso V; Córdoba J; Gobernado M; Carrasco D; Berenguer J; Department of Gastroenterology, La Fe Hospital, Valencia, Spain. Eur J Gastroenterol Hepatol, 1995 Sep, 7:9, 899-903

BACKGROUND: In chronic hepatitis C, relapses of liver disease occur in as many as 50% of patients responding to interferon (IFN) therapy. Although the presence of serum hepatitis C virus (HCV) RNA at the end of therapy predicts a relapse, its absence is not a reliable indicator of cure. In this study we have determined whether responders to IFN (normalization of liver chemistry and clearance of serum HCV-RNA) harbour HCV-RNA in the liver and peripheral blood mononuclear cells (PBMCs), and whether finding the genome at these sites has prognostic significance. METHODS: After the conclusion of therapy, we tested for HCV-RNA in the liver of all the patients (anti-HCV-positive): 16 complete responders with normalization of liver chemistry and clearance of serum HCV-RNA and five non-responders. In 13 of the 16 complete responders we also tested for HCV-RNA in PBMCs. Patients were followed up for 9 months. RESULTS: Liver HCV-RNA was detected in each of the five non-responders and in four of the 16 complete responders (25%). The viral genome was detected in the PBMCs of six complete responders (46%). During follow-up a relapse of hepatitis C occurred in the 10 complete responders with liver or PBMC HCV-RNA but in only one (16%) of the six complete responders without HCV-RNA in liver or PBMCs. CONCLUSION: Reliable information on the prognosis of chronic hepatitis C responders can only be obtained by testing for HCV-RNA in serum, liver and PBMCs at the end of therapy. Patients with HCV-RNA at any of these sites stand a high risk of disease relapse. The risk is low but not abolished in patients without detectable HCV-RNA.

PREDICTION OF RELAPSES AFTER INTERFERON-ALPHA THERAPY BY HEPATITIS C VIRUS RNA IN PERIPHERAL BLOOD MONONUCLEAR CELLS.

Kusaka S; Okusa T; Araki A; Fujiki K; Takashimizu I; Okayasu I; Yamamoto N; Sato C; First Department of Internal Medicine, Faculty of Medicine, Tokyo Medical and Dental University, Japan. J Med Virol, 1995 Jul, 46:3, 265-8

To investigate the predictive value of hepatitis C virus (HCV)-RNA in peripheral blood mononuclear cells in the response to interferon therapy in patients with chronic hepatitis C, 15 patients with histologically proven chronic active hepatitis and who were positive for serum HCV-RNA were treated with interferon-alpha (6 million units; i.m.) every day for two weeks and then three times a week for 22 weeks. Ten of the 15 patients were responders whose alanine aminotransferase levels decreased to the normal range at the end of interferon therapy. In four of the 10 responders, HCV-RNA was not detected by reverse transcription-polymerase chain reaction in peripheral blood mononuclear cells nor in serum at the end of treatment. These four patients were complete responders, with alanine aminotransferase levels remaining normal for the next 24 weeks. In five of the 10 responders, HCV-RNA was detected in peripheral blood mononuclear cells but not in serum at the end of treatment. All of these relapsed within the next 24 weeks. In the remaining responder, HCV-RNA was detected both in peripheral blood mononuclear cells and in serum at the end of treatment. This responder also had a relapse within the next 24 weeks. Five of the 15 patients were non-responders, in whom HCV-RNA was detected both in peripheral blood mononuclear cells and in serum. Thus, detection of HCV-RNA in peripheral blood mononuclear cells may be a good clinical marker to predict relapse after interferon treatment.

HEPATITIS C VIRUS RNA IN PERIPHERAL BLOOD MONONUCLEAR CELLS OF INFECTED PATIENTS BY IN SITU HYBRIDIZATION J Moldvay, P Deny, S Pol, C Brechot and E Lamas; INSERM U-370, CHU Necker, Paris, France.

We used in situ hybridization to detect hepatitis C virus (HCV) infection of peripheral blood mononuclear cells (PBMNC) from 11 patients with chronic active hepatitis. Using 35S-labeled HCV-RNA probe, HCV-RNA-positive and -negative strands were observed in unstimulated PBMNC from three patients, all of whom were receiving immunosuppressive drugs after orthotopic liver transplantation (OLT). HCV-RNA sequences were also identified in PBMNC from three patients who were not undergoing immunosuppression, after stimulation with either phytohemagglutinin (PHA) or pokeweed mitogen (PWM). In contrast, HCV- RNA was not found in the remaining five patients, who had not undergone OLT and whose cells were not stimulated with mitogens. These results show that mononuclear cells can be infected by HCV and that mitogenic stimulation of infected cells increases HCV-RNA replication.

DETECTION OF HEPATITIS B VIRUS DNA IN PERIPHERAL BLOOD MONONUCLEAR CELLS FROM PATIENTS WITH VARIOUS HEPATOPATHIES USING IN SITU HYBRIDIZATION

Suñén E; Fernández de Aranguiz A; De Las Heras B; Gorriño MT; Sarriá L; Malavé C; Campelo C; Cisterna R; Departamento de Microbiología e Inmunología, Universidad del País Vasco UPV/EHU, Hospital Civil de Bilbao, Lejona, Vizcaya. An Med Interna, 1991 Aug, 8:8, 372-6

We have investigated, by "in situ" hibridisation, the presence of hepatitis B virus (HBV) DNA in peripheral blood mononuclear cells (PBMC) from 45 patients with acute and chronic hepatic disorders directly related with HBV or with some seric HBV marker. Results has been related with serological markers and the different types of hepatopaties. The HBV-DNA was detected in PBMC more frequently in patients with hepatic alterations more prolongated (chronic active hepatitis, chronic persistent hepatitis and cirrhosis) than in acute hepatitis patients. It was not detected in any asymptomatic patient with HBV serological markers. As regards HBV serological markers, HBV-DNA was detected in PBMC in 8/11 HBsAg positive patients and in 11/34 HBsAg negative patients: 3 antiHBc positive, 5 antiHBc and antiHBs positive and without conventional seric markers. The detection of HBV-DNA in antiHBc and/or antiHBs positive subjects means the virus may persist after recovery of infection and suggests PMBC could serve as additional reservoirs for reinfection of hepatocytes leading to a reactivation of the liver disease. Our results suggest that HBV infection of PBMC is a frequent event during HBV infection and can have important consequences fundamentally with respect to pathogenic mechanisms of HBV induced liver disease and to the transmission of the virus.

DETECTION OF PRE-S1 PROTEINS IN PERIPHERAL BLOOD MONONUCLEAR CELLS FROM PATIENTS WITH HBV INFECTION

Zoulim F; Vitvitski L; Bouffard P; Pichoud C; Rougier P; Lamelin JP; Trépo C, Hepatitis and AIDS Research Unit, INSERM U271, Lyon, France. J Hepatol, 1991 Mar, 12:2, 150-6

The presence of pre-S1 proteins in peripheral blood mononuclear cell (PBMC) samples from 115 patients with different forms of hepatitis B virus (HBV) infection was investigated by Western blot. Among 67 chronic HBsAg carriers, HBV antigens were detected in the PBMC in 80% for HBsAg, 27% for HBc/e Ag and 34% for pre-S1 proteins. The detection of pre-S1 proteins in PBMC was significantly associated with the presence of serum markers of HBV replication (HBV DNA and/or DNA polymerase). In the group of 48 consecutive patients negative for serum HBsAg, but positive for anti-HBc with or without anti-HBs, HBsAg and pre-S1 proteins could be detected in PBMC. This finding was more frequent among anti-HIV-positive patients (77 and 23% of the cases, respectively) than in the negative ones (23 and 4% of the cases, respectively). The detection of HBV DNA and polyadenylated RNA in some of the PBMC samples positive for HBV proteins suggests that these proteins may be expressed in PBMC, especially during intense HBV replication. In patients negative for serum HBsAg, PBMC may constitute a reservoir of HBV.

PREVALENCE AND SIGNIFICANCE OF HEPATITIS B VIRUS ANTIGENS: EXPRESSION IN PERIPHERAL BLOOD MONONUCLEAR CELLS IN CHRONIC ACTIVE HEPATITIS.

Parvaz P; Lamelin JP; Vitvitski L; Bouffard P; Pichoud C; Cova L; Trepo C; Clin Immunol Immunopathol, 1987 Apr, 43:1, 1-8

One-hundred four chronic active hepatitis (CAH) patients were investigated for the expression of the hepatitis B virus (HBV) surface and core gene products (HBs Ag, HBc/HBe Ags) in peripheral blood mononuclear cells (PBMC). Two-thirds of 59 HBs antigenemic patients expressed HBs Ag in PBMC but 26% of cases positive for both anti-HBs and anti-HBc also expressed HBs Ag while none of the controls reacted. Among HBs antigenemic patients, only those who replicated HBV express the core gene products (HBc and/or HBe Ag) in PBMC, and high replicators did so more often than low replicators (P less than 0.05). The HBs Ag prevalence in PBMC, although slightly higher among HBe Ag/DNAp-positive cases could not be correlated with the intensity of HBV replication. In 16 cases (8 replicants and 8 nonreplicants) HBV DNA was detected by DNA hybridization spot test, while 8 controls devoid of HBV markers were negative. Both T and non-T cells reacted similarly for antigenic or genomic HBV markers. When the expression of HBV gene products in PBMC among 43 cases with HBs antigenemia was compared with that in the liver, a good correlation was found in 70% of cases for HBs Ag but in only 40% for HBc and HBe Ags. By contrast, among 38 cases lacking HBs Ag in the serum but positive for anti-HBc with or without anti-HBs, concordance between liver and PBMC expression of core gene products (69%) was better than for HBs antigenemic patients (40%). These data suggest that PBMC including T lymphocytes may represent the second-best HBV target and may mimic the steps of HBV cycle within hepatocytes.

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