[Changes of humoral and cellular immunity in chronic hepatitis C patients of different staging]. / Izmenenie pokazatelei gumoral'nogo i kletochnogo immuniteta u patsientov s khronicheskim gepatitom C pazlichnoi tiazhesti.

The purpose of the present study was to investigate the influence produced by viral proteins in the hepatic cells and RNA of hepatitis C virus (HCV) on the indices of T- and B-cell response in 52 patients with chronic hepatitis C (CHC). A relative count of peripheral-blood lymphocytes (PBL), expressing antigens CD3+, CD4+, CD8+, CD16+, CD20+ and CD95+ was estimated. The repertoire of antibodies to HCV proteins was specified. The thus obtained data were compared with an activity and a disease stage by using the histological diagnosis and alanine-amino-transferase (ALT) level as well as with the presence of HCV RNA in the serum and viral protein of the liver. Such comparison of data and the use of the correlation analysis made it possible to establish that the antibodies to NS5 protein were detected reliably more often in patients with a more pronounced hepatic fibrosis, with a higher ALT activity and with expression of HCV proteins in the liver. At the same time, the presence of proteins in the liver and of RNA in the serum were accompanied by a more active humoral response to the non-structure proteins of NS4 and NS5 as well as by more profound discrepancies of the immunity T-cell chain (a lowered ratio of CD4+/CD8+ and a smaller content of CD95+). There were no differences between PBL of the studied populations in patients with various activities and an HCV stage. A relatively bigger quantity of CD95(+)--positive PBL was found to be reliably higher in patients with viremia but lower in those cases, in which HCV proteins were detected in the liver. This confirms the inhibiting ability of HCV proteins to the Fas-mediated apoptose of PBL in CHC patient.

Characterization of monoclonal antibodies and epitope mapping of the NS4 protein of hepatitis C virus.

Recombinant DNA containing sequences of HCV NS4 protein was expressed in Escherichia coli cells. Six hybridoma clones producing monoclonal antibodies (MAB) to recombinant NS4 protein (rNS4), aa 1677-1756, were developed. Mapping with a panel of 33 peptides and reciprocal competitive EIA have shown that MAB obtained revealed five antigen determinants, not described earlier: MAB 3F11 and 3F12-one genotype-independent epitope of NS4A (aa 1700-1707) common for genotypes 1, 2 and 3; MAB 1D11-genotype-independent epitope (aa 1713-1728) and MAB 1D3-genotype (subtype 1b)-specific epitope of NS4B (aa 1711-1731); MAB 6B11 and C1-two conformation-dependent determinants in 5-1-1 region. These data indicate that the 5-1-1 region of NS4 protein has a complex antigenic structure and contains at least eight epitopes, including five, revealed in the present work. MAB obtained recognized native viral protein in the cytoplasm of liver cells of patients with chronic hepatitis C. The positive rates of the immunostaining for NS4 antigen using MAB 6B11, 1D11 and 3F12 were 64, 59 and 50%, respectively. It was found that 6B11 MAB to a conformation-dependent epitope much more actively interacts with native NS4 than with the recombinant protein to which MAB was developed. The epitope recognized by 6B11 MAB is highly immunogenic since it induces the B-cell response in all patients investigated with identified anti-NS4 antibodies in blood serum. The MAB panel obtained in this study may become a useful tool for the diagnostic purposes, for the investigation of NS4B function and for the host-viral interactions at the cell level.

[Hepatitis C virus RNA in patients with chronic hepatitis C]. / RNK virusa gepatita C v organizme bol'nykh khronicheskim gepatitom C.

Reviews recent data on the detection of genome and replicative HCV RNA in patients with chronic hepatitis C by PCR and in situ hybridization. Discusses the results of HCV RNA detection in the liver, lymphocytes, serum, and other organs and tissues and notes the relationship between the incidence of RNA and activity of the pathological process. Analyzes the results of HCV RNA detection after IFN treatment. Discusses the role of HCV RNA in the pathogenesis of hepatitis C.

[The role of hepatitis C virus RNA detection in various substrates of patients with chronic hepatitis C]. / Znachenie viiavleniia RNK virusa gepatita C v razlichnykh substratakh u bol'nykh khronicheskim gepatitom C.

Puncture biopsy of the liver and blood count were made in 72 patients with chronic hepatitis C (CHC). Morphological alterations in the liver were assessed by Knodell index. The blood serum, lymphocytes and hepatic tissue were examined for a genome form of hepatitis C virus (HCV) RNA, blood lymphocytes and hepatic tissue--for a relevant replication form. HCV RNA was detected using "nested" RT-PCR. Only 26% patients had symptoms of asthenovegetative and dyspeptic syndromes. Normal alaninaminotransferase (ALT) level was observed in 24% patients, the rest had it high. HCV RNA was encounted more frequently in hepatic tissue than lymphocytes or serum (83, 68 and 46%, respectively). A replication form of HCV RNA was present in hepatic tissue of 31% patients and was absent in the lymphocytes. The incidence of the RNA detection was not related either to the disease symptoms or morphological alterations in hepatic tissue. The occurrence of the genome and replication forms in hepatic tissue does not correlate to ALT level. HCV RNA occurs more often in the serum, blood lymphocytes and in three substrates simultaneously in patients with hyperalatemia.

[Use of PCR for detection of positive (genomic) and negative (replicative) RNA chains of hepatitis C virus in the serum, lymphocytes, and liver of patients with chronic hepatitis C]. / Vyiavlenie pozitivnykh (genomnykh) i negativnykh (replikativnykh) tsepei RNK virusa gepatita C v syvorotke krovi, limfotsitakh i tkani pecheni bol'nykh khronicheskim gepatitom C s pomoshch'iu polimeraznoi tsepnoi reaktsii.

The presence of viral RNA in liver tissue and peripheral blood serum and lymphocytes of patients with chronic hepatitis C (CHC) was studied by polymerase chain reaction with nested primers on the 5'-untranslated region of hepatitis C virus (HCV) genome. Positive (genome) RNA was more often detected in the liver (81% cases) than in the peripheral blood serum (55%) or lymphocytes (64%). Active replication of HCV (presence of negative RNA chains) was observed only in the liver (in 37% cases). Correlation between the frequency of HCV RNA detection in the liver, blood cells and sera and parameters of the inflammatory process activity (SGPT level, histologic activity index and sclerosis index) was investigated. No relationship between the studied parameters was revealed. Positive correlation between the presence of HCV genome RNA in lymphocytes and serum was detected. A tendency to a decrease in the incidence of replicative RNA of the virus in liver tissue with increase in the activity of chronic hepatitis C was observed.