non-B Hepatitis Patients Research Articles

A protracted war of human beings against virus: Interpreting for the Nobel Prize in Physiology or Medicine in 2020

On 5th October of 2020, the Nobel committee announced in the Karolinska Institute that the 2020 Nobel Prize in Physiology or Medicine was awarded to Harvey J. Alter, Michael Houghton, and Charles M. Rice to commend their contribution in the aspect of discovering hepatitis C virus (HCV). Alter and his team acquired C100-3 protein by utilizing recombinant yeast to express viral sequence segments and found the HCV antibody by performing radioimmunoassay. Houghton and his colleagues successfully created the cDNA library from the serum of non-A and non-B hepatitis patients in bacteriophage. Subsequently, the large-scale blind screening method was performed in the serum of patients, acquired clone 81 from the library screening as the clone 5-1-1 involving short sequences to be the probe, and confirmed HCV ultimately. In 1995, Rice and his collaborators completed the full-length HCV genome sequencing from the 3′ terminal sequence using a quantitatively competitive reverse transcription PCR method and verified the full-length sequence has infectivity. This is the first causative agent entirely discovered by molecular biological techniques in human being’s history. Furthermore, Wakita with his team, acquired a full-length HCV genome from acute HCV patients and transformed viral RNA to Hun-7 cell to generate HCV virion in 2005. On this basis, Rice and his team successfully established the cell-cultured system that can effectively generate HCV with infectivity. According to the WHO latest data, there were approximately 71 million chronic HCV cases and nearly 400000 deaths globally in 2016. About 14% of the total HCV infected individuals worldwide lived in China. As a common infectious pathogen globally, HCV infection may cause severe threats and damages to human health, such as chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. The high genetic variation is major characteristics of HCV, which has phylogenetically generated more than 7 genotypes and hundreds of subtypes. The various genotypes and subtypes have distinct geographic distribution and different responses on anti-virus treatment. The resistant mutation for the clinical using drugs including Epclusa (Sofosbuvir and Velpatasvir), is occurring. Moreover, the highly chronic progression render HCV as a silent slayer for human health, and plenty of HCV cases were difficult to be timely diagnosed due to its high proportion of asymptomatic. All these indicated that HCV prevention and control are still facing severe challenges.

Seroepidemiology of hepatitis E virus in patients with non‐A, non‐B, non‐C hepatitis in Hungary

Many cases of acute hepatitis remain undiagnosed and the hepatitis E virus (HEV) is emerging in industrialized countries. The aim of this study was to assess the role HEV as causative agent in acute non-A, non-B, and non-C hepatitis patients in Hungary. 10.5% of the 264 acute non-A, non-B, and non-C hepatitis patients tested had anti-HEV IgG and 1.9% had anti-HEV IgM as tested by ELISA. After confirmation by Western blot 6.1% of the acute non-A, non-B, and non-C hepatitis patients had anti-HEV IgG antibodies only and 1.1% of the patients had both IgG and IgM. All 19 patients that were positive for anti-HEV IgG and/or IgM tested negative for HEV RNA by PCR. Only a small proportion of the acute hepatitis cases in the southwest of Hungary are assumed to be attributed to HEV infection, however, hepatitis E should be considered along with hepatitis A, B, and C in the diagnosis of acute hepatitis.

SEN Virus Prevalence among Non-B and Non-C Hepatitis Patients with High Liver Function Tests in the South of Turkey

SEN Virus Prevalence among Non-B and Non-C Hepatitis Patients with High Liver Function Tests in the South of Turkey

Epidemiology of hepatitis A and E virus infection in Brazil

This review has the objective to discuss the epidemiological aspects of the enterically transmitted hepatitis A and E in Brazil. The prevalence of hepatitis A varies greatly in different Brazilian regions, from 56% in South and Southeast to 93% in North region (Manaus, Amazon). Such differences are also found in different socioeconomic levels among age groups. A significantly higher prevalence was seen in the low socioeconomic group between 1-30 years. This difference is most striking in the first 10 years of age (23.5% vs 60.0%, high/middle vs low, respectively). Despite the improvements in sanitary conditions, hepatitis A is still endemic and outbreaks may occur. As an increasing proportion of the population is becoming susceptible to hepatitis A virus infection and as adult individuals may present more severe forms of the disease, the authors conclude that the implement of hepatitis A vaccination should be considered. Some Brazilian data have shown that the genotype found in our country were IA and IB. Isolates from this study were closely related genetically (or even identical) to isolates originating in other South American countries and overseas, providing firm evidence for epidemiological links between persons who travel to endemic areas. In spite of favorable environmental conditions, outbreaks of hepatitis E have never been reported in Brazil. Nevertheless, reports have demonstrated the evidence of anti-hepatitis E virus antibodies in some Brazilian regions. The seroprevalence of IgG anti-hepatitis E virus among normal populations shows positivities of 6.1% in gold-miners, 3.3% in general population, 2.0-7.5% in blood donors, 1.0% in pregnant women, and 4.5% in children, with no differences among regions. In populations at risk the prevalence of anti-hepatits E virus varies greatly. Among patients with acute non-A, non-B, non-C hepatitis 2.1% was detected in the Southeast to 29% in the Northeast, in 10.6% of acute non-A, non-B, non-C hepatitis relatives in the Amazon basin, in 12% of acute sporadic non-A non-B hepatitis patients in the Northeast, a co-infection with acute hepatitis A in 25 to 38% in the Northeast, in 14 to 18% among prostitutes and women considered at risk for human immunodeficiency virus in the Southeast, and in 12% of the intravenous drug users in the Southeast.

Frequent detection of the human herpesvirus 6-specific genomes in the livers of children with various liver diseases.

This study was performed to investigate the frequency of human herpesvirus 6 (HHV-6) infection of the liver in children with a variety of liver diseases and to evaluate the role of HHV-6 infection in pediatric patients with prolonged non-B non-C hepatitis. Detection of the HHV-6 genomes in liver, in peripheral blood mononuclear cells (PBMC), and in plasma was performed by PCR or by in situ hybridization. Liver biopsy materials from 48 patients, in whom HHV-6 infection was serologically confirmed, were available for PCR analysis. Sequences of the HHV-6B genome were detectable in the livers of 36 of 48 patients (75%). The presence of the genome was not associated with serum transaminase activities. The genome was detectable in PBMC of 22 of 31 (71%) patients tested. In these 31 patients HHV-6 was detected in both the livers and PBMC of 20, was detected in PBMC but not in the livers of 2, was detected in the livers but not in PBMC of 3, and was detected in neither of samples of 6. In situ hybridization of the livers of six patients showed the presence of the HHV-6B genome in the nuclei of hepatocytes. The anti-HHV-6 immunoglobulin M antibody was detectable in 2 of 9 of the non-B non-C hepatitis patients, whereas none of the 22 patients with etiology-defined liver diseases tested positive. Cell-free viral DNA was not detectable in either group of patients. Our results showed that HHV-6B is frequently present in the livers of children with a variety of liver diseases but do not support the assumption that HHV-6B infection of the liver is associated with prolonged non-B non-C hepatitis.

Empty virus-like particle-based enzyme-linked immunosorbent assay for antibodies to hepatitis E virus.

Hepatitis E, an enterically transmitted non-A, non-B hepatitis, is a serious viral infection that occasionally causes large epidemics in developing countries. In developed countries, the disease only appears sporadically due to the transmission routes, and it is considered to be less important. The hepatitis E virus (HEV) cannot grow in cultured cells and no reliable assay system has ever been developed. In addition, the present diagnostic are not perfect, and actual rates of HEV infection may be underestimated. Highly purified empty virus-like particles (VLPs) of HEV have been produced by the use of a recombinant baculovirus vector in insect cells. Using these VLPs as an antigen, an enzyme-linked immunosorbent assay (ELISA) for antibodies to HEV was developed. A panel of 164 sera that were randomized and coded, and sera collected periodically from three patients with hepatitis E were used for the evaluation. The sensitivity of the assay was shown to be equal to or better than that obtained in previous research that used the same serum panel. The ELISA demonstrated that the serum IgM level of the patients was highest at the onset of the clinical illness and then rapidly decreased. In contrast, a high level of circulating IgG antibody titers lasted for more than 4 years. In Japan, a non-endemic country, the prevalence of the IgG class antibody to HEV in healthy individuals was found to range from 1.9% to 14.1%, depending on the geographical area. Only one out of 900 (0.1%) serum samples was IgM-positive. The IgM class antibody to HEV was detected in 10.8% of non-A, non-B, and non-C acute hepatitis patients in northeast China, whereas none of the patients in Korea had the IgM antibody. The ELISA utilizing the VLPs is sensitive and specific in its detection of the IgM and IgG antibodies to HEV. The ELISA is therefore useful for diagnosing HEV infection and for seroepidemiological study of hepatitis E.

Prevalence of GBV-C/hepatitis G virus viraemia among blood donors, health care personnel, chronic non-B non-C hepatitis, chronic hepatitis C and hemodialysis patients in Egypt

A new RNA virus, designated GBV-C/hepatitis G virus (HGV) has been identified recently. To evaluate the prevalence of GBV-C/HGV infection among Egyptians, five groups were enrolled in this study: group I, healthy blood donors (82); group II, health care personnel (30); group III, chronic non-B non-C hepatitis patients (63); group IV, chronic hepatitis C patients (100); group V, renal dialysis patients (79). GBV-C/HGV-RNA was detected by nested reverse transcription-polymerase chain reaction (RT-PCR) using primers derived from 5′-non coding region of GBV-C/HGV. GBV-C/HGV-RNA was detected in 57 of 354 tested sera with an overall prevalence of 16.1%. Meanwhile, isolated GBV-C/HGV infection was detected in 16/57 (28.1%), GBV-C/HGV coinfection with hepatitis C virus (HCV) in 37/57 (64.9%) and with hepatitis B virus (HBV) in 4/57 (7.6%) of cases. The highest prevalence was encountered among dialysis patients reaching 30% followed by chronic hepatitis C (14%), blood donors (12.2%), chronic non-B non-C hepatitis (11.1%), whereas the lowest prevalence rate of 6.6% was detected among health care personnel. Nucleotide sequence analysis in three Egyptians confirmed that these PCR products were derived from GBV-C/HGV genome and all isolates classified into US/European type (type 2) of GBV-C/HGV genotypes. The risk factors of all cases were non-transfusion parenteral exposure, e.g. reusing syringes, dental treatment, surgery, invasive medical maneuvers, with an exception of renal dialysis patients who have had repeated blood transfusion. It is concluded that there is a relatively high prevalence of GBV-C/HGV-RNA among different Egyptian groups compared to international figures. The main risk factors were direct percutaneous exposure rather than blood transfusion. The Egyptian GBV-C/HGV isolates are very similar to the American isolate PNF 2161.

Serum Neopterin Assay to Interpret the Main Cause of Chronic Liver Damage Involved with Plural Etiological Factors

Summary Identification of the main cause of chronic liver damage (CLD) involved with plural etiological factors is important for the management and the treatment of patients. To evaluate its clinical usefulness in interpreting the main cause of CLD involved with hepatitis C virus, alcoholic overdrinking, or fatty steatosis, which are the most frequent and troublesome etiological facotors in the daily outclinic, serum neopterin level was measured in ten healthy subjects, ten chronic hepatitis C (CH-C) patients, nine alcohol-induced liver damge (AL) patients, eight non-alcholic fatty liver (FL) patients, eight CH-C with AL padtients, seven CH-C with FL patients and ten non-B non-C chronic hepatitis (CH-NBNC) patients by using radioimmunoaasay. There were no significant difierences in the mean serum neopterin levels among the healthy subjects, the AL patients and the FL patients. The serum neopterin level in the CH-C with AL or FL patients was significantly higher than that in the AL or the FL patients and just as high as that in the CH-C patients. Furthermore, serum neopterin level in CH-NBNC patients was as high as that in the patients with CH-C. Therefore, the serum neopterin assay can easily assume the main cause of CLD involved with plural causes, so it will be very useful in the daily clinical practice as an easyto- use auxiliary test.

HCV and HBV coexist in HBsAg-negative patients with HCV viraemia: possibility of coinfection in these patients must be considered in HBV-high endemic area.

Hepatocellular carcinoma (HCC) is one of the most common cancers and is highly associated with hepatitis B virus (HBV) infection in Korea. The role of HBV and hepatitis C virus (HCV) in HCC patients who are negative for hepatitis B surface antigens (HBsAg) remains poorly defined. It has been suggested that HCV core protein may impair the polymerase activity of HBV in vitro, potentially lowering HBV titre in coinfected patients. Therefore, routine enzyme immunoassay may not detect HBV, in spite of the presence of HBV viraemia in low titres. The aim of this study was to confirm the coexistence of HBV viraemia in hepatitis C-infected patients with HCC who have apparent HBsAg seronegativity and to establish the need for clinical reinterpretation of enzyme immunoassay (EIA) serological tests of HBsAg in patients with HCV viraemia and HCC. The serological profiles of HBV and HCV in 616 patients with HCC were analysed and the coinfection rate of HCV and HBV investigated. Sera were obtained from 16 patients who were both anti-HCV and HCV-RNA positive but HBsAg negative, and tested for HBV by polymerase chain reaction (PCR). Eleven non-A and non-B chronic hepatitis patients without HCC who had the same profiles of anti-HCV, HCV-RNA, and HBsAg were tested for HBV by PCR. As a control group, sera were obtained from 15 patients with HCC and 30 non-A and non-B chronic hepatitis patients without HCC; both were anti-HCV, HCV-RNA, and HBsAg negative and tested for HBV PCR. Of the 616 patients with HCC, 450 (73.1%) had current HBV infection, 48 (7.8%) had anti-HCV antibodies, and nine (1.5%) had viral markers of both HCV and HBV by serological profiles. Of the 27 patients with HCV viraemia and HBsAg seronegativity (16 with HCC; 11 with non-A non-B chronic hepatitis), 14 (51.9%) showed HBV viraemia by PCR. In contrast, of the 75 patients in the control group (45 with HCC; 30 with non-A and non-B chronic hepatitis) who were both HCV PCR negative and HBsAg negative, five (11.1%) showed HBV viraemia by PCR. The PCR for HBV revealed coexistent HBV viraemia in HCV viraemia patients, despite HBsAg negativity by EIA. In HBV-endemic areas, the possibility of coinfection of HBV in HBsAg-negative patients with HCV viraemia should be considered and molecular analysis for HBV-DNA performed.

Plasma levels and hepatic mRNA expression of transforming growth factor-β 1 in patients with fulminant hepatic failure

Background/Aims: Transforming growth factor-β 1 is an important cytokine involved in cell growth and inflammation which has been shown to be inhibitory to hepatic DNA synthesis. The aim of this study was to investigate the plasma levels and hepatic mRNA expression of transforming growth factor-β 1 ni patients with fulminant hepatic failure in whom liver regeneration may be impaired. Methods: Plasma levels of transforming growth factor-β 1 and human hepatocyte growth factor were measured in 57 fulminant hepatic failure patients and 20 healthy volunteers by ELISA. Northern blot analysis of transforming growth factor-β 1 and H3 histone, a marker for liver proliferation, was performed in liver tissue of 14 fulminant hepatic failure patients. Results: The plasma levels of total transforming growth factor-β 1 in fulminant hepatic failure patients on admission (median 38.8 ng/ml, range 8.4–108 ng/ml) were significantly higher than those in control subjects (23.0 ng/ml, 8.5–34.9 ng/ml, p<0.001). Significantly higher levels were observed in non-A, non-B hepatitis patients (57.9 ng/ml, 38.8–108 ng/ml, n=10, p<0.001) compared to patients with paracetamol overdose (37.1 ng/ml, 8.4–72.5 ng/ml, n=47). In contrast, the plasma levels of free transforming growth factor β 1 were greater in paracetamol overdose (623 pg/ml, 46.7–1241 pg/ml, n=21) than in non-A, non-B hepatitis (131 pg/ml, 77.2–254 pg/ml, n=9), with both being higher than control (72.3 pg/ml, 28.7–108, n=7, p<0.001). The plasma levels of human hepatocyte growth factor in patients with paracetamol overdose (7.04 ng/ml, 1.00–62.4 ng/ml) were significantly higher than those in patients with non-A, non-B hepatitis (4.48 ng/ml, 0.74–9.10 ng/ml, p<0.05). Northern blots showed increased mRNA expression of transforming growth factor-β 1 in paracetamol-overdose patients ( n=8, p<0.05), but not in patients with non-A non-B hepatitis ( n=6), compared to controls ( n=4). Conclusions: The increased circulating plasma TGF-β 1 in FHF may be part of the tissue repair process in fulminant hepatic failure. In patients with non-A, non-B hepatitis, the increased total transforming growth factor-β 1 together with a less elevated hepatocyte growth factor could be related to impaired liver regeneration in this group.

Hepatitis E virus infection in a cohort of patients with acute non-A, non-B hepatitis

Background/Aims: The aim of this study was to determine the frequency of hepatitis E virus infection in a cohort of patients with acute non-A, non-B hepatitis in Greece. Methods: Serial serum samples of 198 patients with acute non-A, non-B hepatitis and a single serum specimen from 316 healthy subjects were tested for IgG and IgM antibodies to hepatitis E virus (anti-HEV). Results: Anti-HEV IgG was found in 15 198 (7.6%) of acute non-A, non-B hepatitis patients and 7 316 (2.2%) of healthy controls ( p=0.007). Anti-HEV IgM was found in 2 198 (1.0%) acute non-A, non-B hepatitis patients and in none of the healthy subjects. Neither anti-HEV IgM (+) case reported any risk factor and neither had travelled in areas endemic for hepatitis E virus infection. HEV-RNA was detected by reverse transcription polymerase chain reaction in one patient. The prevalence of anti-HEV IgG was 7 45 (15.6%), 1 46 (2.2%), 5 30 (16.7%) and 2 77 (2.6%) in acute non-A, non-B hepatitis reporting transfusion, intravenous drug use, occupational/hospitalization, and unknown transmission, respectively ( p=0.007). Anti-HEV IgG was found in 13 122 (10.7%) and 2 76 (2.6%) of acute non-A, non-B hepatitis patients positive and negative for anti-HCV, respectively ( p=0.03). A similar association was found with anti-HBc ( p=0.007). The prevalence of anti-HEV IgG was significantly higher in cases reporting transfusion [OR=7.3, 95% C.I. 1.4–37.7, p=0.017] and occupational/hospitalization [OR=6.8, 95% C.I. 1.2–38.2, p=0.029], as transmission category after controlling for age. Conclusions: These findings indicate that: (a) hepatitis E virus may be a cause - although not a frequent one - of sporadic or community-acquired acute non-A, non-B hepatitis in Greece; (b) hepatitis E virus may share transmission routes with hepatitis B and C viruses; and (c) the hypothesis that hepatitis E virus may be transmitted by parenteral routes deserves careful consideration.

The distribution of hepatitis C virus genotypes in Turkish patients.

The distribution of hepatitis C virus (HCV) genotypes was investigated in 89 HCV-infected Turkish patients. Blood samples were collected from haemodialysis patients (n = 45), chronic liver disease (CLD) patients (n = 38), acute non-A, non-B (NANB) hepatitis patients (n = 2) and blood donors (n = 4). HCV RNA sequences were amplified in the 5' non-coding region and were typed by restriction fragment length polymorphism analysis. The predominant genotype was 1b (75.3%), followed by 1a (19.1%), 2 (3.4%) and 4 (2.2%). While there was no significant difference in the distribution of HCV genotypes with respect to age, sex, transfusion history, alanine aminotransferase levels or liver histology (in the CLD group), type 1a-infected patients were younger than type 1b-infected patients (P < 0.05) in the haemodialysis group. Serological reactivity to recombinant HCV proteins was assessed in 58 samples using the Chiron RIBA-2 assay. The reactivity of samples from patients infected with type 1b with 5-1-1 and c100 antigens was significantly lower (P < 0.05) than the reactivity of samples from those infected with type 1a. These results, together with the results of two previous studies, indicate that HCV genotypes 1, 2, 3 and 4 are prevalent in different frequencies in the Turkish population. Determination of the genotype distribution of HCV in a geographical area may provide important clues for studying the epidemiology, transmission and pathogenesis of HCV-related diseases and may also aid in improving serological assays to detect HCV infection.

Serological evidence for hepatitis E virus infection in Israel.

Israel is suspected to be endemic for hepatitis E virus (HEV) because of its geographic location and the large-scale immigration from endemic countries. Although no cases of local HEV infection have been diagnosed, a serological survey would provide indirect evidence for such infection. We examined sera from 1,416 healthy subjects, including 1,139 Jews from various regions of Israel and 277 Arabs, most of whom reside in the West Bank of the Jordan River. In addition, we tested 13 non-A, non-B, and non-C viral hepatitis patients. Sera were screened for antibody to hepatitis E virus (anti-HEV) by a newly developed enzyme immunoassay (EIA) and by immunoblots for both IgG and IgM anti-HEV activity. Positive samples were confirmed by neutralization. The seroprevalence found by EIA was 2.81% and 1.81% in the Jewish and Arab populations, respectively. More than a 2-fold higher prevalence in males compared to females and an increase with age were found in both populations. However, these differences were nonsignificant. The geographical distribution was even throughout the country, except for two clusters of 3 and 4 seropositive individuals possibly reflecting past foci of infection. Eight of 37 EIA-positive sera were positive for IgG, and 3 were positive for IgM by the immunoblot assay. Among hepatitis patients (9 acute and 4 chronic), one patient with chronic hepatitis was positive for both IgG and IgM. Our study provides indirect evidence that Israel is endemic for HEV.(ABSTRACT TRUNCATED AT 250 WORDS)

Sensitivity and Specificity of Three Third-Generation Anti-Hepatitis C Virus ELISAs

Three commercially available 3rd-generation anti-HCV ELISAs (Abbott, Murex and Ortho) were evaluated in various serum panels: (A) blood donor samples (n = 403) with 1st- or 2nd-generation anti-HCV ELISA (various manufacturers) positive test results; (B) non-A, non-B hepatitis patients (n = 212); (C) multi- transfused patients (n = 253); (D) serial dilutions of HCV confirmed (RIBA and PCR) positive blood donors (n = 24), and (E) first-time blood donors (n = 1,055). All samples of panels A, B and C were tested in PCR and RIBA-2. In panels A, B and C, 398 samples were HCV PCR positive: all were detected by Abbott and Ortho, and 397 (99.7%) by Murex. The sample missed by the Murex ELISA showed an isolated anti-C33c reactivity in RIBA-2. In panels A-C, 442 samples were RIBA-2 positive and all were detected by the 3 tests. With Probit analysis on results of panel D, no significant difference in sensitivity was observed between the 3 evaluated ELISAs. Specificities of Abbott, Murex and Ortho in 1,055 blood donors were 99.7, 99.3 and 99.9%, respectively (NS, χ2). We conclude that the sensitivity and specificity of the 3 ELISAs are comparable although the C33c antigen in the Murex VK47 test should be improved.

Isolation of the novel agent from human stool samples that is associated with sporadic non-A, non-B hepatitis.

The agent(s) responsible for sporadic non-A, non-B hepatitis in humans was serially transmitted in rhesus monkeys by intravenous inoculation of the stool extract from a patient. A novel agent called HFV (hepatitis French [origin] virus) was present as 27- to 37-nm particles in the infectious stool extract. Hepatopathic lesions were noticed in infected monkeys during the acute phase of illness. The purified viral 27- to 37-nm particles consist of a double-stranded DNA of approximately 20 kb and are detected in infected monkey liver. Analysis of cell culture detects the approximately 20-kb-long viral DNA in stool samples from infected monkeys and sporadic enteric non-A, non-B hepatitis patients. Furthermore, the 27- to 37-nm viral particles were able to protect monkeys challenged with infectious stool extract. Our results indicate that 27- to 37-nm virus like particles are responsible for sporadic non-A, non-B hepatitis in rhesus monkeys.

Detection of hepatitis C virus antigen by immuno-histochemical staining: a histological marker of hepatitis C virus infection

Hepatitis C virus has been recognized as a major cause of non-A, non-B viral hepatitis. Although serologic tests have been commercialized, no specific histological or immuno-histochemical markers for hepatitis C virus infection are available for routine use. In an effort to detect hepatitis C virus antigen in liver tissue we investigated the immuno-reactivity to monoclonal antibodies on frozen liver tissue from a chimpanzee and patients with chronic non A, non B hepatitis. Monoclonal antibodies were developed in mice immunized with a synthetic peptide derived from hepatitis C virus core antigen. One monoclonal antibody was reactive and showed typical cytoplasmic granules in chimpanzee hepatocytes. Using this monoclonal antibody a similar staining pattern was found in the liver biopsies of 21 out of 28 chronic non-A, non-B hepatitis patients, positive for hepatitis C virus-RNA and anti-HCV. The granular immuno-reactivity was abolished after pre-incubation of this monoclonal antibody with infected chimpanzee liver or with hepatitis C virus synthetic peptide but not with normal chimpanzee or human liver tissue. There was no reactivity in four patients with hepatitis C virus-RNA-negative, anti-HCV-positive chronic non-A, non-B hepatitis, in 11 patients with chronic type B hepatitis or in 12 hepatitis C virus-RNA-negative, anti-HCV-negative patients with various liver diseases. However, staining was found in three out of four additional chronic type B hepatitis patients suspected of co-infection with non-A, non-B agents.(ABSTRACT TRUNCATED AT 250 WORDS)

Recombinant immunoblot assay reaction patterns and hepatitis C virus RNA in blood donors and non‐A, non‐B hepatitis patients

To establish the value of the second-generation recombinant immunoblot assay (RIBA-2) and cDNA polymerase chain reaction (cDNA PCR) for confirmation of hepatitis C virus (HCV) infection, anti-HCV reaction patterns and the presence of HCV RNA were examined in 610 blood donors and 255 non-A, non-B hepatitis patients who were positive or indeterminate in RIBA-2. Of RIBA-2-positive donors (n = 191) and patients (n = 224), 75.4 and 89.7 percent, respectively, were HCV RNA positive. The most frequently observed anti-HCV recognition patterns in HCV RNA-positive donors and patients were c22, c33c, and c100 and/or 5-1-1 (67.3%, 57.7%) and c22, c33c (24.8%, 29.3%). Among subjects with a RIBA-2-indeterminate result, HCV RNA was detected more often in patients (n = 31) than in donors (n = 419): 67.7 and 2.1 percent, respectively. In viremic persons with single-band reactivity in RIBA-2, this reactivity was always directed against either c22 or c33c. HCV RNA was detected by cDNA PCR in none of 162 persons with only anti-c100 and/or anti-5-1-1 reactivity. Therefore, RIBA-2 reactivity against c100 in combination with 5-1-1 should not be considered positive but indeterminate. In RIBA-2-indeterminate persons, HCV RNA detection is necessary for reliable confirmation of HCV infection.

Antibodies to hepatitis C virus and hepatitis C virus infection

The authors studied the interrelationship of antibody to hepatitis C virus core protein (anti-HCV core), antibody to C100-3 antigen (anti-C100-3) and serum hepatitis C virus RNA positivity in chronic liver disease patients. Anti-HCVcore was detected with high titers in 95% (69/73) of chronic non-A, non-B hepatitis patients, while anti-C100-3 was found in 60% (44/73). A close relationship was observed between detection of anti-HCVcore and viremia. Anti-HCVcore was also detected with low titers in 24% (10/41) of hepatitis B virus carriers negative for anti-C100-3. Hepatitis C virus RNA was found in 3 of the 10 anti-HCVcore-positive carriers. A significant correlation was observed between the occurrence of high titers of anti-HCVcore and serum hepatitis C virus RNA positivity. In chronic hepatitis C patients treated with interferon-alpha, a sustained reduction of anti-HCVcore was more closely associated with sustained virus clearance than that of anti-C100-3. These results indicate that anti-HCVcore may serve as a reliable marker of hepatitis C virus infection.

Aplastic anemia and hepatitis C: Molecular biology exonerates another suspect

Objective. - To test the hypothesis that the rare, often fatal, syndrome of hepatitis-associated aplasia is associated with hepatitis C virus infection. Design. - Case series. Setting.-Tertiary referral centers in the United States, Japan, Italy, and Germany. Patients. - Twenty-eight patients with onset of aplastic anemia within 90 days after seeking medical attention for jaundice, or having serum transaminase levels 150% or more of normal (hepatitis-associated aplasia patients) and three patients who developed aplastic anemia following liver transplantation for non-A, non-B, hepatitis. Outcome Measures. - Presence of hepatitis C in serum, bone marrow, and liver samples, detected by the polymerase chain reaction; antibody testing; and percentage of activated peripheral cytotoxic T lymphocytes determined by immunophenotyping. Results. - Hepatitis ribonucleic acid was present in the serum samples of 10 (36%) patients with hepatitis-associated aplasia. However, hepatitic C virus viremia was associated with transfusions received after the onset of aplasia: seven (58%) of 12 patients with hepatitis-associated aplasia who had received 21 or more units of blood products at the time of serum sampling were viremic, compared with only three (19%) of 16 patients with hepatitis-associated aplasia who had received 20 or less units of blood products (P &lt;.05). Hepatitis C virus was not found in blood and bone marrow samples of three National Institutes of Health case patients tested at the time of diagnosis. None of three livers from non-A, non-B hepatitis patients who developed aplastic anemia after liver transplantation contained hepatitis C virus ribonucleic acid. Activated CD8+ T lymphocytes were elevated three- to 20-fold early in the course of hepatitis-associated aplasia. Conclusions. - Our results implicate a novel, non-A, non-B, and non-C agent in both hepatitis-associated aplasia and fulminant hepatitis.

Diagnosis of hepatitis C virus (HCV) infection using an immunodominant chimeric polyprotein to capture circulating antibodies: reevaluation of the role of HCV in liver disease.

Structural and nonstructural regions of the HCV-encoded polyprotein have been expressed in recombinant yeast, bacteria, or insect cells and used to capture and measure reactive antibodies circulating in different individuals. The putative nucleocapsid protein (C) and nonstructural proteins 3-5 (NS3-NS5) were found to contain the most immunodominant epitopes. The NS3, NS4, and C regions were expressed in yeast in the form of a fused, chimeric polyprotein (C25) and a capture assay for reactive antibody was developed. This anti-C25 assay detects all previously identified HCV-seropositive cases and provides a substantially more sensitive diagnostic for both acute and chronic HCV infections than the current anti-C100-3 (NS4) assay. Anti-C25 was detected more frequently than anti-C100-3 in chronic, transfusion-associated non-A, non-B hepatitis patients from the United States (95% vs. 71%) and Japan (98% vs. 82%), in cryptogenic cirrhosis patients from the United States (62% vs. 28%), and in hepatitis B surface antigen-negative cases of hepatocellular carcinoma from Japan (83% vs. 63%). These data indicate that HCV has a greater role in these liver diseases than was previously thought. In volunteer United States blood donors sampled following the introduction of anti-C100-3 screening, the prevalence of anti-C25 and anti-C100-3 was 0.5% and 0.08%, respectively.