Acute non-A, non-B hepatitis--clinical, epidemiological and histological characteristics

Among 73 consecutive patients with biopsy documented acute non-toxic hepatitis, half of the patients (49%) had acute type B hepatitis, while 27 patients (37%) had acute type A infection. One patient had a significant rise in antibodies against cytomegalovirus. The remaining 10 patients (14%) fulfilled the criteria of hepatitis type non-A, non-B. The main type of exposure for hepatitis A was visit to endemic hepatitis areas (41%), and for type B it was drug addiction (46%). Half of the patients with hepatitis non-A, non-B had no known hepatitis exposure while some had visited endemic hepatitis areas or were drug addicts. The patients with non-A, non-B hepatitis had significantly less biochemical changes as compared to the patients with hepatitis B. In contrast, the histological findings showed the greatest activity in the biopsies from patients with hepatitis B and non-A, non-B. Follow-up liver biopsies in half of the patients with non-A, non-B hepatitis showed no signs of chronic active liver disease. It is concluded that hepatitis type non-A, non-B is a significant problem in Denmark.     

The chronic sequelae of non-A, non-B hepatitis.

Twenty-six of 388 patients (6.7%) followed prospectively after open-heart surgery developed non-A, non-B hepatitis. Of these 26, 12 had an elevated (often fluctuating) serum alanine aminotransferase (SGPT) for greater than 1 year. Liver biopsy, done in eight of 12, showed chronic active hepatitis in six and chronic persistent hepatitis in two; one patient with chronic active hepatitis had early cirrhosis. Anicteric patients with peak SGPT greater then 300 IU/L were at greatest risk of developing chronic hepatitis. Chronic non-A, non-B hepatitis was symptomatically mild and unaccompanied by physical signs or laboratory evidence of autoimmune disease or severe chronic liver disease. In all 12 patients there was spontaneous improvement in serum transaminase over a period of 1 to 3 years, and four patients had sustained normalization of SGPT. Thus chronic active hepatitis is a common sequela of acute non-A, non-B hepatitis but may have a better prognosis than chronic active hepatitis of other causes.     

Role of hepatitis C virus in non-B chronic liver disease.

To assess the contribution of the recently identified hepatitis C virus to chronic liver diseases of unknown cause and chronic hepatitis attributed by exclusion to non-A, non-B hepatitis, we tested for antibody to hepatitis C in hepatitis B surface antigen-negative patients with a spectrum of chronic liver diseases. Antibody to hepatitis C virus, a marker of hepatitis C infection, was detected with a first-generation radioimmunoassay at the following frequencies in the following patient groups: 69% of transfusion-associated non-A, non-B hepatitis; 53% of non-transfusion-associated non-A, non-B hepatitis; 26% of hepatitis B surface antigen-negative hepatocellular carcinoma; 8% of cryptogenic cirrhosis; 5% to 7% of autoimmune chronic liver diseases; 19% of patients with miscellaneous types of chronic liver disease; and 0.67% of healthy controls. Among non-transfusion-associated cases, 81% with a history of intravenous drug use but only 18% with occupational exposure as health workers had antibody to hepatitis C virus. Among cases of hepatocellular carcinoma, 63% of Japanese patients but only 11% of American patients had evidence of hepatitis C infection. Comparison in a subgroup of 79 serum samples of a second-generation radioimmunoassay with the first-generation assay demonstrated a 12% increase in antibody frequency from 30% to 42%. We conclude that hepatitis C plays a substantial role in transfusion-associated and non-transfusion-associated non-A, non-B hepatitis as well as in hepatocellular carcinoma, especially in Japan, a limited role in cryptogenic cirrhosis, and essentially no role in autoimmune chronic liver diseases. Application of more sensitive immunoassays will increase the frequency of antibody seropositivity in all subgroups, but relative distinctions among risk groups are likely to remain.     

Post-transfusional vs. sporadic non-A, non-B chronic hepatitis. A clinico-pathological and evolutive study

The clinical, morphological and evolutive features of 60 patients with chronic hepatitis, presumably caused by non-A, non-B virus infection, have been retrospectively analyzed. In all the cases the disease began as an acute episode of viral hepatitis that was followed by persistently abnormal liver function tests. No patient had evidence of current or past hepatitis B virus infection and other known causes of chronic liver disease were excluded. Thirty patients had received blood transfusions in the recent past, five were drug addicts and the source of the infection was not identified in the remaining 25, in whom the disease was considered to be sporadic. Clinical or biochemical differences between patients with post-transfusional and sporadic non-A, non-B chronic hepatitis were not observed, but liver histology showed a higher proportion of patients with chronic persistent hepatitis in the sporadic (72%) than in the transfusional group (53%). On follow-up, sustained normalization of liver function tests was observed in 46% of the cases with sporadic hepatitis but only in 13% of the cases with post-transfusion hepatitis. These observations suggest that non-A, non-B chronic hepatitis is more severe in patients with transfusion-related infection than in sporadic cases.     

The role of acute hepatitis type A, B, and non-A non-B in the development of chronic active liver disease

In 19 patients followed from biopsy-verified acute viral hepatitis to chronic active liver disease and 74 patients followed to complete resolution verified by a normal liver biopsy, sera from the acute phase were studied for serologic evidence of hepatitis type A and B. Eleven of the 19 patients who developed chronic active liver disease progressed from acute hepatitis type B and 7 from acute hepatitis type non-A non-B. One patient could not be classified because the sera were exhausted. None had serological markers of actual hepatitis type A infection. Of the 74 patients with a histologically complete resolution, the acute episode could be classified as type B hepatitis in 47 and type A hepatitis in 13 patients. The remaining 14 patients were classified as having acute viral hepatitis type non-A non-B. Our findings confirm that type B and non-A non-B hepatitis may give rise to chronic liver disease, whereas type A hepatitis so far has not been demonstrated to initiate a chronic liver disease.     

Non-A, non-B hepatitis after open-heart surgery in Sweden

In recent years evidence has emerged that most post-transfusion hepatitis is caused by one or more previously unknown agents named non-A, non-B. A prospective investigation was made of 74 patients who underwent open-heart surgery. Only volunteer blood was used for transfusions. Transfusion-associated hepatitis appeared in 15 (20%) of the patients 4-12 weeks after the operation. In no case was the hepatitis found to be caused by hepatitis B, A or Epstein-Barr virus. One patient had a cytomegalovirus infection; the other 14 cases (19%) were classified by definition as non-A, non-B hepatitis. Although most of the patients were asymptomatic and all were anicteric, the course of the hepatitis was protracted in many cases. Thus, 6/12 observed patients still had pathologic transferase values more than a year after the onset of hepatitis. Liver biopsy was performed in 3 cases and showed histologic signs of chronic active hepatitis in all of them.     

The long-term prognosis of non-transfusion-associated non-A, non-B hepatitis. A clinical, epidemiological, and histological investigation

In a prospective study of the natural course of acute hepatitis, 157 of 1020 patients with biopsy-verified acute hepatitis could be classified as having hepatitis type non-A, non-B. We here report on the long-term prognosis for these 157 patients. The main type of exposure was drug addiction (40%), whereas 40% had no known hepatitis exposure. Only two patients had received blood products (blood transfusion and factor VIII). Follow-up liver biopsy (mean histological follow-up, 22 months) in 94 of the 157 patients showed chronic liver disease in 15-that is, cirrhosis in 6, suspicion of cirrhosis in 2, chronic aggressive hepatitis in 5, and chronic persistent hepatitis in 2. There was a striking predominance of elderly women with no known hepatitis exposure and with a high frequency of autoantibodies in serum among the patients with progression to chronicity, whereas chronic non-A, non-B hepatitis in drug addicts or after blood transfusions seems to be a limited problem. A comparison of histological features in the initial biopsies from patients with progression to chronicity or complete resolution showed piecemeal necrosis and abnormal bile duct epithelium to be of prognostic value.     

Detection of hepatitis C viral RNA sequences in fresh and paraffin-embedded liver biopsy specimens of non-A, non-B hepatitis patients.

In this study methods of HCV-RNA detection in fresh frozen and formalin-fixed, paraffin-embedded liver biopsies are described. Of 22 untreated chronic non-A, non-B hepatitis patients and 6 control patients, a plasma sample and part of a liver biopsy were freshly frozen for hepatitis C virus (HCV) cDNA-PCR. From 16 of the same non-A, non-B hepatitis patients and from 5 of the same control patients formalin-fixed, paraffin-embedded liver tissue from the same biopsy was available also for HCV cDNA-PCR. In 13 of 22 non-A, non-B hepatitis patients HCV-RNA could be detected in plasma as well as in liver tissue. In the other 9 non-A, non-B hepatitis patients and in 6 control patients, no HCV-RNA was detectable in either plasma or liver tissue. The comparison between HCV cDNA-PCR results in fresh frozen versus formalin-fixed, paraffin-embedded liver biopsies showed that although detection of HCV-RNA in both correlated 100% the quantity of HCV-RNA was lower in the formalin-fixed, paraffin-embedded liver biopsies of 5 of 8 patients for whom end-point dilution titration of liver RNA was performed. We conclude that using the procedures described HCV-RNA can be reliably detected in both fresh-frozen and formalin-fixed, paraffin-embedded liver biopsies and that HCV cDNA-PCR in liver tissue may become an important assay, especially for monitoring anti-viral therapy.     

Chronic Viral Hepatitis in Thalassemic Liver Disease: A Long-Term Study in Patients with Acute Hepatitis with Nontransfusion-Dependent Thalassemia Minor

To evaluate the effective role of hepatitis viruses in thalassemic (Th) liver disease, we carried out a long-term study in 42 subjects with nontransfusion-dependent Th minor hospitalized for an episode of acute viral hepatitis. 10 patients had serologic evidence of hepatitis A, 23 of hepatitis B and 9 of hepatitis non-A, non-B. In the follow-up chronic hepatitis was detected histologically in 5/23 patients with hepatitis B and 5/9 with hepatitis non-A, non-B. All hepatitis A patients recovered completely. The prevalence in 7 out of 10 patients with chronic hepatitis of piecemeal necrosis and of inflammatory changes over hepatic siderosis and fibrosis evidenced a determinant role of chronic viral infection in the development of liver damage in these patients. Thus, heterozygous nontransfusion-dependent Th patients seem to have a high risk of developing a chronic inflammatory liver disease especially after an episode of non-A, non-B hepatitis. Therefore, in our geographical area, chronic hepatitis of viral origin should be taken into account, among other pathogenetic factors, in many cases of cryptogenic thalassemic liver disease.     

Long-term analysis of non-A, non-B hepatitis--clinical, histologic and immunologic findings

By means of a long-term analysis of histological and clinical-biochemical data taken of a group of patients (n = 38) having contracted an acute viral hepatitis non-A, non-B with a uniform parenteral source of infection evidence is given concerning the dynamic processes in the course of chronic non-A, non-B hepatitis. The analysis was therefore not based on the total biopsy material but on a section of 143 liver biopsies of patients who had been punctured at least 3 times (up to 6 times) in the 7 years of examination. A rate of 57,2% chronic persistent and 42,8% chronic lobular hepatitis was ascertained. A chronic active hepatitis or liver cirrhosis were not found. Both the histological activity degrees and ALAT-mean values were declining during the follow-up study though attacks (ALAT) and histological reactivations were observed in the 7th year, too. The data suggest that the chronic liver changes that developed in this type of non-A, non-B hepatitis proved to be extremely lingering but as a whole a regressive tendency could be observed.     

Detection of hepatitis B virus DNA in paraffin-embedded liver tissues in chronic hepatitis B or non-A, non-B, hepatitis using the polymerase chain reaction

We developed a polymerase chain reaction assay for the direct detection of hepatitis B virus in paraffin-embedded liver tissue and applied this assay to determine whether hepatitis B virus DNA exists in livers with chronic hepatitis non-A, non-B. Fifty five liver biopsy samples were studied: 11 from patients with HBeAg-positive chronic hepatitis (paraffin-embedded) and 44 from patients with chronic hepatitis non-A, non-B (21 paraffin-embedded; 25 fresh frozen). Thirty three (75%) of the non-A, non-B cases were positive for hepatitis C virus antibodies. Approximately 1 to 10 ng of DNA was extracted from the paraffin-embedded tissue and amplified using oligonucleotide (23-mer) primers specific for the S gene (positions 261 to 692). The beta-globin gene was used as an internal control for sensitivity because this is a single copy gene and allows for relative quantification. In each of the chronic hepatitis B livers, the expected 432-base-pair amplification product for hepatitis B virus DNA and beta-globin gene product were both detected. On the other hand, in the 21 paraffin-embedded chronic hepatitis non-A, non-B livers, no hepatitis B virus DNA was detected, although beta-globin gene was observed in all. Furthermore, in all 25 frozen non-A, non-B livers, beta-globin gene was observed, but no hepatitis B virus band was seen. The limit of detection of hepatitis B virus DNA by this method was estimated to be one genomic copy of hepatitis B virus DNA per cell.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pseudotumor appearance in chronic hepatitis

A pseudotumor appearance of the liver has not been previously described in chronic hepatitis. We reviewed 81 charts with a biopsy diagnosis of "chronic hepatitis" and at least one radiologic liver scan, and found four cases with a pseudotumor appearance. They all had features on radiologic scans suggestive of space-occupying lesions. Histology showed chronic hepatitis in all four. Two were cirrhotic, one due to alcohol and the other due to hepatitis B. The other two patients had chronic active hepatitis with regenerative nodules but only limited fibrosis, one due to hepatitis B, the other probable non-A, non-B. None had any evidence of hepatic malignancy. We conclude that some patients with chronic hepatitis may present with a pseudotumor appearance on radiological scans, due to the presence of regenerative nodules. Space-occupying lesions on liver scans in chronic hepatitis may represent non-neoplastic liver disease.     

Long-term follow-up of chronic post-transfusion non-A, non-B hepatitis: clinical and histological outcome

A chart review of chronic hepatitis cases at the Department of Infectious Diseases, Roslagstull Hospital, Karolinska Institute, Stockholm, Sweden, revealed 37 patients with chronic non-A, non-B hepatitis, caused by blood transfusions or intravenous gammaglobulin infusions. They had been followed up long-term, mean 46 months (range 10-149). During the initial hepatitis episode most patients had been anicteric and 13/37 (35%) asymptomatic. Yet, the majority developed progressive liver disease with chronic active hepatitis, with or without histological signs of cirrhosis. Thus, 10/18 (56%) on whom a liver biopsy had been performed within 7-12 months had chronic active hepatitis, four (22%) with histological signs of cirrhosis, and of the five patients biopsied after greater than or equal to 5 years of follow-up, three (60%) had histological signs of cirrhosis. Accordingly, as the duration of follow-up increases, an increasing number of patients with post-transfusion non-A, non-B hepatitis seem to develop cirrhosis.     

Satellite vasculitis of B or non-A non-B hepatitis. Diagnostic value of a provocation test by intradermal injection of histamine

The histo-pathological lesions of hypersensitivity vasculitis (HV) consist in an inflammatory reaction to circulating immune complexes deposited on the vascular endothelium. A provocation test by intradermal injection of histamine was used to demonstrate HV in patients with chronic hepatitis. Three groups of patients were studied: 16 with chronic hepatitis and HV, 22 with chronic hepatitis without HV, 4 control subjects. Skin biopsy was performed before and 3 hours after intradermal histamine. The following markers were looked for in the serum, skin and liver of all patients with chronic hepatitis: HBV (HBs Ag and HBe Ag, anti-HBs, anti-HBe and anti-HBc in the serum, HBs and HBc Ag in the liver); non-A non-B (non-A, non-B Ag and antibody, anti non-A non-Bc in the serum, non-A, non-Bc Ag in the liver). Viral antigens were found in healthy skin in 2 cases (1 chronic A hepatitis, 1 vasculitis), and after injection of histamine in 9 cases (4 chronic A hepatitis, 5 vasculitis). Viral antigens were found on skin biopsy in 2 patients with negative sera. Significant skin changes after histamine were detected only in HV (75 p. 100) even in the absence of cutaneous vasculitis (6/12 positive cases). An intradermal histamine provocation test is a simple, non-invasive method for diagnosing HV.     

Hepatitis C

The major cause of chronic post-transfusion hepatitis, the hepatitis C virus (HCV), has been identified. HCV is a single-stranded linear RNA virus with characteristics similar to the flaviviruses. A different agent, the hepatitis E virus, is associated with epidemic (enterically-transmitted) non-A, non-B hepatitis. At present, infection with HCV is recognized by the finding of anti-HCV antibodies, positive in up to 90% of patients with chronic non-A, non-B post-transfusion hepatitis. Antibodies to HCV are detected in 1% of normal volunteer blood donors and in the majority of donors implicated in post-transfusion hepatitis. HCV antibodies are also found in patients with autoimmune liver disease and hepatocellular carcinoma. Moreover, HCV infection may contribute to the pathogenesis of liver disease in alcoholic patients. The role of HCV infection in fulminant non-A, non-B hepatitis and hepatitis-associated aplastic anemia has not been elucidated as yet. Therapy of chronic non-A, non-B hepatitis with recombinant human alpha-interferon has been shown to improve or normalize aminotransferase levels in approximately 50% of patients, most of whom have evidence of HCV infection. However, relapse after cessation of treatment is common. In the future, screening blood for evidence of HCV infection may prevent most cases of non-A, non-B post-transfusion hepatitis.     

Electron microscopic evidence of non-A, non-B hepatitis markers and virus-like particles in immunocompromised humans

Characteristic pathological alterations of the liver in chimpanzees inoculated with non-A, non-B hepatitis sera have been described, but no corresponding findings have been reported in humans. Electron microscopic studies of the liver biopsy specimens of two homosexual patients with acquired immune deficiency syndrome, one without hepatitis (Patient 1) and one with chronic active hepatitis in remission (Patient 2), revealed the cytoplasmic tubular structures which are characteristic of chimpanzee non-A, non-B hepatitis. A cluster of 23 nm double-shelled particles was also seen in the cytoplasm of a hepatocyte in patient 1 who had received a blood transfusion 8 days before the biopsy. These particles were smaller than the Dane particles, Epstein-Barr virus, cytomegalovirus or herpes simplex virus, and different from hepatitis A virus particles; the antibodies to all of which are found in high concentration in acquired immune deficiency syndrome patients. These observations may reflect the morphologic findings for non-A, non-B hepatitis infection in humans.     

Epidemic non-A, non-B hepatitis in patients from Pakistan

Epidemic non-A, non-B hepatitis was diagnosed in three young Pakistani men during a 10-month period at the Los Angeles County-University of Southern California Medical Center. All three patients had recently visited or lived in Karachi, Pakistan. None had serologic markers of hepatitis B virus infection or IgM antibody (acute-phase) to hepatitis A virus. A liver biopsy from one patient showed marked cholestasis and cholangiolar transformation of hepatocytes, a pattern previously described in patients with epidemic non-A, non-B hepatitis. Immune electron microscopy of a stool specimen obtained from this patient 10 days after the onset of symptoms showed virus-like particles, 27 nm in diameter, that were specifically aggregated by antibody contained in acute-phase sera from the three Pakistani patients, from patients with non-A, non-B hepatitis in Burma and Nepal, and from an experimentally infected marmoset. Recognition of three separate cases of probable epidemic-type non-A, non-B hepatitis in patients at one institution during such a short time suggests that Pakistan is endemic for this infection and that the disease may be more commonly spread to the United States than is now presumed.     

Detection by immunofluorescence of an antigen-antibody system in patients with acute and chronic non-A,non-B hepatitis

ABSTRACT— An antigen-antibody system has been identified by immunofluorescence in patients with non-A, non-B hepatitis. The non-A, non-B antigen was localized in the hepatocyte nuclei of liver biopsies from patients with acute post-transfusion or sporadic non-A, non-B hepatitis and in those from patients with chronic post-transfusion non-A, non-B hepatitis, the percentage of positive cells being most prominent in patients receiving immunosuppressive treatment. Absence of the antigen in normal livers and in livers from patients with type B hepatitis infection indicated its specific association with non-A, non-B infection. Antibody reacting with the nuclear antigen became detectable in serum during post-transfusion acute non-A, non-B hepatitis in 11 out of 15 cases; it was absent before transfusion. Six out of 12 cases of sporadic acute non-A, non-B hepatitis were also found to produce the antibody, which was repeatedly found to be absent during the acute phase in five patients with type A and in eight with type B hepatitis. The non-A, non-B antibody, mainly an IgM antibody, persisted in serum for prolonged periods of time after onset, both in patients showing biochemical resolution of their illness and in those who continued to have liver damage after the acute phase. Accordingly, eight out of nine patients withchronic non-A, non-B hepatitis were found positive for the antibody in serum, seven at the time the non-A, non-B antigen was detected in their liver. Thus this non-A, non-B associated antigen-antibody system shares remarkable similarities of behaviour with the “core” system of the hepatitis B virus.     

Detection of 2',5' oligoadenylate synthetase activity in acute viral hepatitis with special reference to histologic features in the acute stage.

We measured the sequential changes in 2',5' oligoadenylate synthetase activity in 21 patients with acute viral hepatitis (5 with type A, 6 with type B, and 10 with type non-A, non-B hepatitis) by radioimmunoassay. Liver biopsies were performed during the acute phase in all patients. Among patients with acute hepatitis A and B, the 2',5' oligoadenylate synthetase levels were transiently elevated at the time of the peak alanine aminotransferase level in the patients in whom a liver biopsy showed acute hepatitis or non-specific reactive hepatitis. Of 10 patients with acute non-A, non-B hepatitis, 4 showed a similar 2',5' oligoadenylate synthetase activity pattern and liver histology to those observed in acute hepatitis A and B. In the remaining 6, the 2',5' oligoadenylate synthetase levels remained elevated for 3.5 to 6 months while the alanine aminotransferase was elevated. Liver biopsy in these patients showed chronic hepatitis. Persistent detection of raised 2',5' oligoadenylate synthetase activity levels during the acute stage of non-A, non-B hepatitis may thus be an indicator of progression of the disease.     

Immunophenotyping of lymphocytes in liver tissue of patients with chronic liver diseases by flow cytometry.

Immunological factors are important in the pathogenesis of a spectrum of hepatobiliary diseases. To characterize the nature of specific immunological responses in liver disease, we determined lymphocyte changes in liver tissue and in blood using flow cytometry. A total of 113 liver biopsy specimens was collected from patients with the following diseases: 19 chronic hepatitis B; 39 chronic non-A, non-B hepatitis; 27 alcoholic liver disease; 10 hepatic malignancy; 8 autoimmune hepatitis; 6 fatty liver and 4 primary biliary cirrhosis. The lymphocytes were isolated from the liver biopsy specimens by mechanical and enzymatic methods. The lymphocyte yield was 7,901 +/- 575 cells/mg of liver tissue. The viability of lymphocytes was 97.7% +/- 0.3%. Lymphocytes were stained with four pairs of two-color mixed fluorescein-conjugated monoclonal antibodies, including T4-T8 (CD4/CD8), T11-B1 (CD2-CD20), NKH1-T8 (CD56-CD8), IL-2R1-T11 (CD25-CD2), and the ratios were determined by an Epics Profile flow cytometer. Immunophenotyping of lymphocytes in whole blood samples was simultaneously analyzed. Variability in lymphocyte yield and different patterns of lymphocyte subsets were found in the liver biopsy specimens. The yields of lymphocytes from patients with chronic non-A, non-B and autoimmune hepatitis were highest, and the lowest yield was from patients with fatty liver. Patients with primary biliary cirrhosis, fatty liver and hepatic malignancy had relatively high ratios of CD4/CD8, CD56/CD8 and CD25/CD2; whereas patients with chronic hepatitis B, autoimmune hepatitis and non-A, non-B hepatitis had lower ratios of CD4/CD8, CD56/CD8 and CD25/CD2. No difference in lymphocyte ratios between the patients with cirrhotic and noncirrhotic alcoholic liver disease was found.(ABSTRACT TRUNCATED AT 250 WORDS)