Detection of transfusion-associated hepatitis caused by non-A, non-B, non-C flavivirus.

Sera of patients suffering from acute hepatitis, and different forms of chronic hepatitis were found to be reactive to reagents prepared from the yellow fever virus (YF) vaccine strain. Serum samples of 1974 patients were tested, and 133 of them were positive. Hepatitis C virus specific antibodies were absent from the majority of them. The frequency of antibodies to other flaviviruses (tick-borne encephalitis, West Nile) and hepatitis B virus markers was similar to that measured among the population in Hungary positive for any of the surrogate markers of hepatitis infections. Results of both immunofluorescence tests, and Western blots suggest that there is a non-A, non-B, non-C hepatitis virus circulating among the Hungarian population, which possesses antigenic cross-reactivity with the yellow fever virus, but the identity to any of the known flaviviruses could not be verified yet. No history of yellow fever vaccination could be revealed in any of the patients included into this study. The anamnestic data on previous transfusions or surgical operations can be verified only in the case of the half of YFV-positive patients, nevertheless, the sexual transmission seems to be very infrequent. Attempts are continued in order to detect the viral RNA using polymerase chain reaction, and clone cDNA sequences for sequence analysis.

[Advances in the diagnosis of non-A non-B viral hepatitis]. / A non-A, non-B virushepatitis diagnosztikájának fejlödése.

Having constructed the serological diagnostics of Hepatitis A and B viruses there remained non identified agent(s): named "Non-A, non-B". The non-A, non-B infecting parenterally is a Flavivirus, with lipoid envelope and a diameter of 30-60 nm, as stated very early. After having known the simple filament RNS genome, named as Hepatitis C. In the same time it was tried to isolate the antigen and antibody from ill people, without success. There were not usable the surrogate markers too (anti-HBc and ALT). One part of the synthetized polypeptides from little fragments of C virus genome was apt to antibody-detection. In the acute state there was 20-40, in the chronic state 60-80% anti-HCV positive during the illness. This seropositivity is not equal of healing, neither the potency to infect. The infectivity is showed by the nucleic acid's presence (of the virus) in the blood (PCR), the perfect healing by the neutralizing antibody.

[Transfusion-associated non-A, non-B, non-C hepatitis caused by flaviviruses]. / A non-A, non-B, non-C flavivírus által okozott inokulációs hepatitis.

Hepatitis C virus was shown to be a member of the flavivirus family. Tick-borne encephalitis virus and West Nile virus, members of the same family occur in Hungary, too. Serum samples from patients suffering from transfusion associated hepatitis were tested with yellow fever virus antigens for specific IgG, and IgM using immunofluorescence test. Eight hundred serum samples were tested. Yellow fever virus related IgG antibodies were found in 232 sera. In the case of 72 patients specific IgM antibodies could also be detected. The majority of the IgM positive patients underwent surgical operation and/or blood transfusion 1 to 2 months before the onset of the disease. Fifty-four sera positive for yellow fever virus-related antibodies were tested with HCV reagents, but only 13 were found to be positive, or cross-reacting. The 20 patients with yellow fever related antibodies were controlled with tick-borne encephalitis antigens, too. Nevertheless, no measurable cross-reaction could be detected. No measurable cross-reaction could be detected with the West Nile virus. The hepatitis B markers also were tested in 44 sera positive for yellow fever antibodies. There was only one, which contained HBsAg, and 10 of them proved to be positive for anti-HBcAg. The results indicate, that a non-A, non-B, non-C flavivirus is also present in the Hungarian population, which can be detected on the basis of the antigenic cross-reactivity with the attenuated yellow fever virus. This virus seems to be responsible for every 11th transfusion associated hepatitis examined.(ABSTRACT TRUNCATED AT 250 WORDS)

Prevalence of antibody to hepatitis C virus in blood donors, high-risk groups and patients with liver diseases in Hungary. A multicentre study using ABBOTT EIA test and a comparison with an ORTHO ELISA test system.

Serum samples from 1185 individuals (blood donors, health-care workers, patients on haemodialysis, those from other high-risk groups and those suffering from non-A, non-B hepatitis or other liver diseases) were examined for antibody to a recombinant HCV antigen. An ABBOTT HCV EIA system was used throughout and in addition a parallel study with ORTHO HCV ELISA was done in 380 of the samples to compare the two anti-HCV tests. A confirmatory neutralizing ABBOTT ELISA probe was also performed in 45 cases. The anti-HCV test was positive in 1.60% of the healthy blood donors and in 9% of subjects excluded from donation for elevated aminotransferase. In patients on haemodialysis 47%, in other high-risk-group subjects 33% anti-HCV prevalence was found. Patients with acute and chronic post-transfusion NANB hepatitis showed 40% and 70% prevalence, respectively. The two ELISA tests revealed 95% agreement in the parallel determinations. Serial end-point-dilution studies of anti-HCV-positive sera suggest that the ABBOTT test was of superior sensitivity. The results of the confirmatory test suggest that reactive (positive) samples of low optical density near to the cut-off value, required a confirmation with the naturalization test. HCV infection seems to be a common aetiological factor in PT-NANB hepatitis in Hungary, therefore, screening of blood donors for anti-HCV may be justified.

[Preparation of human heart valve grafts in Hungary]. / Human szívbillentyü transzplantátumok készítése a hazai gyakorlatban.

A method of preparation and preservation of valved aortic and pulmonary allografts is described, used in corrective procedures of congenital heart diseases. The method of sterilization, exclusion of bacterial and viral contaminations is enhanced. Complications did not occur after implantations of the grafts.

HBsAg-like structures in immunosuppressed mice inoculated with human hepatitis B virus.

Thymectomised and irradiated DBA/2 mice were injected intraperitoneally with human serum containing high titer of HBsAg, and were positive for HBsAg. Through the entire experiment neither degenerative and inflammatory lesions nor hepatitis B virus antigens could be detected in the liver of these animals by histomorphology and immunofluorescence, respectively. The sera of all these mice were negative for HBsAg by radioimmunoassay. By electron microscopy, however, increasing amounts of filaments and round particles measuring 20-22 nm in diameter could be observed in the endoplasmic reticulum of the mouse hepatocytes from the 8th day following injection. From the 90th day after inoculation the number of the filaments increased in an extreme degree. After fixation with KMnO4 and EDTA preferential staining, the filaments proved to be highly electrondense. According to the authors the filaments observed in mouse livers are lipoproteins produced by the hepatocytes in response to HBV inoculation. The appearance of the filaments is HBsAg-like, though their immunological characteristics become modified.

Occurrence of tubuloreticular inclusions in acute viral hepatitis.

Light and electron microscopy were used to study the liver needle-biopsy material of 20 patients with acute viral hepatitis. According to clinical and serologic data, 5 cases proved to be acute viral hepatitis type A, 13 were type B, and 2 were type non-A/non-B. In 2 of the hepatitis type A, in 6 of the hepatitis type B, and in one of the type non-A/non-B cases, tubuloreticular inclusions (TRI) were found in the endoplasmic reticulum of the sinusoidal endothelial cells of the liver. These data support the possible presumption that the inclusions represent a characteristic reaction of the endoplasmic reticulum to different influences, as for example to viral infection, rather than to the virus itself.

Cytochemical characterization of hepatitis B virus infected liver cells in chronic liver diseases.

The cytochemical characteristics of the hepatitis B virus (HBV) components, the nuclei and cytoplasm of the infected cells were studied in liver needle biopsy material obtained from patients suffering from serologically HBsAg positive chronic hepatitis. The HBV surface and core components were found to be more resistant to proteolytic enzymes and phospholipase than the other cellular components, and only combined digestion was effective. With preferential RNP and specific DNP staining the accumulation of granules 10-20 nm and 20-40 nm in diameter could be observed in the liver cell nuclei. The cytochemical-morphological polymorphism of the latter was well detectable. Part of the granules corresponded to accumulated perichromatin granules. It is assumed that the groups of granules 20-40 nm in diameter, showing a varying density on DNP staining, similar to the chromatin on RNP staining, and not surrounded by a peripheral area represents one of the forms of appearance of the HB virus core component.

Serologic studies of hepatitis viruses for their aetiologic role in chronic liver disease.

Serum samples from 130 patients with chronic (persistent or active) hepatitis, including cirrhosis, and 70 patients with chronic alcoholic disease, all confirmed by biopsy, were examined by counter electrophoresis (CEP) for HBsAg. The patients were residents in the South-Transdanubian region of Hungary. Of the former 25%, of the latter 3%, proved positive. In 138 of the cases, 4 different tests were performed, viz. CEP, complement fixation, reverse passive haemagglutination (RPH) and radioimmunoassay (RIA). The results were positive in 24, 36, 43 and 60%, respectively, in chronic hepatitis and in 4, 6, 8 and 14%, respectively, in cases of alcoholic origin. Serum samples from the same patients were tested for anti-HBs and anti-HBc. Positive anti-HBs was found in 24% of the patients with chronic hepatitis and in 26% of those with alcoholic liver disease. The presence of anti-HBc was demonstrated in 42% and 38%, respectively. Joint evaluation of RPH-positive and anti-HBc positive cases on the one hand and of RIA-positive and anti-HBc positive cases on the other resulted in 68% and 71% HB virus-related positivity, respectively. The present study failed to furnish evidence in support of the presence of hepatitis A virus of its involvement in the aetiology of chronic liver disease.

Levamisole treatment of acute viral hepatitis and HBsAG-positive chronic active hepatitis.

A fundamental role is attributed to the pathological immune response in the development of chronic hepatitis B. By virtue of its non-specific immune modulatory effect, levamisole is capable of improving impaired T-cell function. Hence, studies with treated and control groups were performed in determine the effect of levamisole in acute viral hepatitis and chronic active hepatitis B. In acute hepatitis B, levamisole promoted the normalization of GPT, the elimination of HBsAg; this improvement was preceded by higher GPT values and increased titres of IgG, IgA, and of anti-HBcore. In chronic active hepatitis in the first few months of treatment a moderate increase of the GPT and HBsAg levels occurred, followed later by a decrease of these values. At the same time the phytohaemagglutinin reactivity of lymphocytes increased, and so did the ratio of circulating active T-cells.

Progressive vaccinia: immunological aspects and transfer factor therapy.

Progressive vaccinia is a rare and serious complication of smallpox vaccination. Depressed immune function can generally be found as an underlying disorder; thus adequate immuno-correction may be expected to be therapeutically effective. Humoral and cell-mediated immunity was repeatedly examined in one case throughout the course of the disease. Results indicated partial deficiency of cell-mediated immunity. No therapeutic effect was achieved by using human antivaccinia immunoglobulin and N-methylisatin beta-thiosemicarbazone. Transfer factor therapy was also attempted. Treatment with a non-specific transfer factor preparation was followed by a transitory clinical improvement. A specific transfer factor preparation given during the last month of life, however, had no therapeutic effect. The patient died on the 145th day after vaccination. Autopsy findings pointed to combined immune deficiency.