Genetic subtypes of HIV type 1 in Hungary.

We examined the diversity of HIV-1 subtypes in 11 adults from Hungary, using the heteroduplex mobility assay (HMA) and DNA sequencing. HMA results showed that HIV-1 gp120 sequences from 10 patients were of subtype B, whereas 1 patient, infected in Africa, carried a subtype C strain. DNA sequencing confirmed the HMA results and revealed a high intrasubtype diversity in the C2V3 region of env in different clade B isolates, which suggests multiple introduction of subtype B to Hungary. This study shows that subtype B is the predominant HIV-1 clade in Hungary.

Periodic reappearance of bovine herpesvirus type 4 DNA in the sera of naturally and experimentally infected rabbits and calves.

A BHV-4 specific nested PCR was used for the detection of viral DNA in serum samples of rabbits and calves. All animals were followed up for 62 days, blood samples were taken for PCR studies every second day. Maternal infection of calves resulted in the repeated regular reappearance (10-14 days) of the virus (DNA) in serum samples. When PCR positive five-day-old calves were infected with tissue culture adapted virus, the reappearance of the DNA in the serum was shown to be irregular, nevertheless, DNA peaks reappeared during the whole observation period. A PCR negative calf infected at the age of 60 days was found to possess viraemia until p.i.d. 32. In rabbits treated intravenously with BHV-4 the inoculum or a primary viraemia was detected at p.i.d. 2-3 and p.i.d. 14-16. Published data on human herpesviruses suggest, that the target cells might be a pluripotent stem cell population of the bone marrow and differentiated virus-infected cells destroyed by the immune system might be the source of viral DNA detected in the serum. Frequency of DNA reappearance was depended on the age of the infected animals but not on the inoculated amount of BHV-4. The described phenomenon might be part of BHV-4 infection of very young animals.

Human herpesvirus 8 in hematologic diseases.

Human herpesvirus type 8 (HHV-8), also known as Kaposi's sarcoma-associated herpesvirus (KSHV) is a new member of the g-herpesvirus family. It is an unusual herpesvirus in that it carries a large number of genes that encode oncoproteins or cell signaling proteins. In addition to being the causative agent of both HIV-associated and non-HIV-associated Kaposi's sarcoma this DNA tumor virus has been implicated in the pathogenesis of several diseases. These include multiple myeloma (MM), Waldenstöm's macroglobulinemia (WM), multicentric Castleman's disease (MCD), body cavity-based lymphoma (BCBL), and various other conditions such as sarcoidosis and pemphigus. While the causative role of the viral infection is fairly certain in the development of BCBL and multicentric Castleman's disease, HHV-8 may act through a different mechanism to induce plasma cell malignancies. It has been suggested though the finding is still controversial - that infection of bone marrow stromal dendritic cells by HHV-8 might be a key factor in the etiology and pathogenesis of monoclonal gammopathies. The aim of this review is to provide a short introduction into the tumorigenic potential of HHV-8 as well as to detail the available data and possible mechanisms on the involvement of this virus in different hematologic diseases.

Spontaneous lymphoma in an AKR mouse with dominance of 42 chromosomes.

The chromosome content of the lymphoma cells derived from various organ manifestations of an AKR female mouse with spontaneous lymphoma was investigated. It was found that the lymphoma cells were heterogeneous and the dominant subpopulation of the lymphoma cells contained 42 chromosomes. At least 8 subpopulations of the lymphoma were detected at the karyotype analysis, from which the 41XX; +15; 41XX; +18; 42XX; +15, +18 and 44XX; +5, +8, +15, +18 karyotypes were the most frequent aberrations. The frequency of the presence of various subpopulations was different also in the thymus, spleen and lymph nodes. The authors suggest that over the trisomy of chromosomes 15, other trisomies (e.g. gain of chromosome 18) can play a role in the AKR mouse lymphomagenesis.

Comparative chromosome examination of AKR mouse lymphoma after its i.p. transplantation with thymus- or spleen-derived lymphoma cells into hybrid mice.

A spontaneous AKR female mouse lymphoma was transplanted with its thymus cell suspension (10(6) cells i.p.) into AKR female mice. Lymph node cell suspension derived from one of the leukemic mice was injected (10(6) cells i.p.) into AKR females. The lymphoma cells "homing" to the thymus of one of the AKR females were suspended in Parker solution and 5 X 10(6) cells were given i.p. 5 (C3H X AKR) F3 hybrid mice (group 1). The lymphoma cells "homing" to the spleen of the same AKR female were also suspended, and 5 X 10(6) cells were injected i.p. into 5 (C3H X AKR) F3 hybrids (group 2). Chromosomes were prepared from the thymus and the spleen of two mice in both groups. The karyotypes derived from the hybrids of group 1 were compared to that of the group 2. It was found that the lymphoma cells "homing" to the thymus could be characterized by the trisomy of chromosome 15, while the lymphoma cells "homing" to the spleen had primarily the trisomy of chromosome 18. The results indicate that the thymus manifestation of the spontaneous AKR lymphoma is heterogeneous, and it contains at least two major subpopulations of the lymphoma cells.

Prospects for the control of AIDS patients by introducing defective-HIV harbouring leukocytes.

Introduction of leukocytes harbouring an artificially constructed defective HIV provirus into AIDS patients may result in inducing superinfection resistance against HIV and interfering with HIV receptors or replication of HIV. All these may slow down progression of the disease.

Restriction site maps of the human adenovirus type 8 DNA.

Restriction site maps of human adenovirus 8 (Ad h 8) were constructed with BamHI, HindIII, PstI, SalI and KpnI endonucleases. The genome size was found to be 22.1 to 22.3 X 10(6) Mr. Comparison of the results with the data available on h Ad subgenera A, B, C showed that the SalI enzyme revealed subgenus-specific differences in the genomes. Similar patterns of the SalI fragments in both type 8 and 10 suggest that the differences were specific for the subgenus D of h Ad.

Characterisation of an echovirus type 11' (prime) epidemic strain causing haemorrhagic syndrome in newborn babies in Hungary.

Echovirus 11' (prime) isolates from an epidemic of haemorrhagic syndrome in departments of obstetrics in Hungary have been characterised. The leading component of the clinical disease was carditis and its lethal outcome occurred in 13 newborn babies. Maternal immunity was found to be absent even in women of 41 years of age. The application of monovalent oral poliovirus type 1 vaccine prevented the progress of the epidemic within two weeks. Nevertheless, a serological survey among primovacinees of 3-15 months of age revealed that 20% of the babies seroconverted without clinical symptoms during the epidemic. Serological evidence showed that the echovirus 11' infection was unable to interfere with the efficacy of oral poliovirus vaccine (OPV), since seroconversion rates of primovaccinees did not differ significantly from those in the group seroconverted also to echovirus 11' during the vaccination campaign. A 440 nucleotide (nt) fragment of the 5'-non-translated region of 12 epidemic echovirus 11' isolates and 26 echovirus prototype strains was amplified by a nested reverse transcription-polymerase chain reaction (RT-PCR) and analysed using three different restriction endonucleases. The 5'-regions of the echovirus 11' isolates were found to be identical to each other but different from that of the prototype echovirus 11 (Gregory) strain. The results indicate that echovirus 11' isolates underwent genetic changes in the 5'-end and P1 region of the genome before the onset of the epidemic.

Effects of a herbicide on the peroxide metabolism enzymes and lipid peroxidation in carp fish (Hypophthalmichthys molitrix).

A study was made of the peroxide metabolism enzymes in carp fish, and of the effects on these of three different concentrations of the herbicide paraquat. Mainly the changes in the superoxide dismutase, catalase and lipid peroxidation resembled the changes observed in previous intoxication studies on mammals.

[Human cytomegalovirus, its significance in immune deficiency states, laboratory diagnosis, therapeutic possibilities]. / Humán cytomegalovirus, jelentösége immunhiányos állapotokban, laboratóriumi diagnózis, terápiás lehetöségek.

The authors report on the virological findings of 59 transplant recipients. The following procedures were used for the detection of human cytomegalovirus (HCMV) infection: detection of antiviral antibodies by ELISA, the detection of virus-coded antigens in the patients' leucocytes (HCMV antigenemia test), "accelerated" virus isolation using immunofluorescence (IF). Serial examinations revealed the HCMV infection in 12 patients following organ transplantation. The antigenemia test proved to be positive in all cases. Two third of the cases suffered from viremia. The virus specific serology possess diagnostic value only in every second acute illness. Since the antigenemia test used to be successful in the earliest phase of acute illnesses, the chance of effective chemotherapy can be increased significantly. The virus serological examinations are of essential importance during the pretransplantation screening of donors and recipients. The "accelerated" procedure of virus isolation experiments indicates the presence of infective HCMV within 1 to 4 days. Transplant recipients obtain new life perspectives, nevertheless, the modern diagnostic procedures may only support the prevention of life-threatening virus infections under the conditions of immunosuppression. The excellent mutual cooperation of the clinicians and diagnostic virologists seems to be at least as important condition of successful transplantation medicine as the high technology in surgery and clinical diagnostics.

[Clinical immunological features and interferon therapy in chronic hepatitis C]. / Klinikai immunológiai megfigyelések és interferon terápia krónikus C hepatitisben.

Clinical and immunological findings of 74 patients with chronic hepatitis C have been reported and experiences with interferon-alpha treatment of 31 patients are summarized. In addition, the first results of anti-HCV screening of blood donors are also briefly described. Transfusion in the history was noted in 69% of patients and the time, elapsed from the transfusion to the diagnosis was a mean of 7.15 +/- 8.1 years. Concerning the severity of the liver disease, chronic persistent hepatitis was established in 40%, active hepatitis in 45% and cirrhosis in 15% of the patients, respectively. Cholestasis was recorded in 32% of the cases. A significant elevation of serum immunoglobulin levels was noted in 83%, an antibody to liver specific protein (anti-LSP) has occurred in 80%, cryoglobulinaemia in 44% and circulating immune complexes in 33% of the patients. Natural killer cell activity of peripheral blood mononuclear cells significantly decreased. HLA B8 and DR3 antigens were found with elevated frequency (36.6% and 42.1%). Recombinant interferon-alpha at a weekly dose of 3MU thrice, for six months, has normalized serum alanine aminotransferase in 45% of patients and a sustained remission was found in 26%. The treatment resulted in the clearance of HCV-RNS from the serum in 40% of patients and that well correlated with the complete remission. In the good responders, a decrease in CD4+ cell count and a moderate decrease in CD8+ cell count as well as a transient rise in B cell count were seen during the treatment. Mitogen-induced lymphoproliferative response and natural killer cell activity increased. Predictors of response were as follows: female sex, shorter time elapsed from transfusion, absence of HLA, A1, B8, DR3 and serum anti-HBc negativity. Anti-HCV has been found in 0.33--0.38% of blood donors screened, and it is suggested, that a liver disease accompanied with elevated serum alanine aminotransferase, may be present in about 25-30% of anti-HCV positive symptom-free persons.

[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)

Variability of the PreS1/PreS2/S regions of hepatitis B virus in Hungary.

Infection with the hepatitis B virus can occur perinatally, parenterally, or sexually, and it can cause acute or chronic liver diseases. Phylogenetic analysis of the virus has led to its classification into eight genotypes (A-H), which show a characteristic worldwide distribution. The aim of this study was to reveal the HBV genotypes present in Hungary and to investigate a nosocomial and an intrafamilial outbreak. The collected samples were tested by nested PCR, and a 650-nucleotide-long segment of the preS1/preS2/S region was sequenced. As no previous genotype data were available from Hungary, sera of 24 HBsAg-positive patients were collected from different regions of the country. They also served as control samples for the molecular epidemiologic study. Nineteen of them carried genotype D of hepatitis B virus, and five of them carried genotype A. Twenty-nine patients from a haemato-oncology unit were affected in a nosocomial outbreak. The patients had haematological and/or oncological diseases, most of them were immunosuppressed. In twenty-eight cases, based on phylogenetic analysis of the viruses, there was presumably a common source of infection, and an epidemiological investigation showed that the infections seemed to be hospital-acquired. In the intrafamilial outbreak, two asymptomatic carrier children infected their foster mother. The three sequences were totally identical.

A soluble factor(s) released by MRC-5 cells early and late after human cytomegalovirus infection induces maturation of monocyte-derived dendritic cells.

A human cytomegalovirus (HCMV) strain passaged 10 times on MRC-5 human fibroblast cells failed to express immediate early (IE) antigens in immature dendritic cells (iDCs) after infection. However, both the early and the late HCMV conditioning medium, harvested from MRC-5 cells at 24 h or 7-9 days after infection, respectively, induced a higher ratio of DCs expressing maturation markers (CD40, CD83, CD86 and HLA-DR) on the surface of the cells. HCMV conditioning medium, ultracentrifuged to remove virus particles, exhibited a similarly enhanced expression of DC maturation markers. DCs treated with HCMV conditioning medium harvested late after infection increased the percentages of autologous CD4+ and CD8+ cells of seropositive donors to produce IFN-gamma and stimulated HCMV-specific lymphoproliferative responses. The early HCMC conditioning medium was also able to induce the functional maturation of DCs, as demonstrated by supplementing this medium with a Chlamydia pneumoniae antigen.

Genotypic distribution of human herpesvirus-8 strains circulating in HIV-positive patients with and without Kaposi's sarcoma in Hungary.

Open reading frame (ORF) 26 of human herpesvirus-8 (HHV-8) from peripheral blood samples of 15 Hungarian HIV-positive patients with or without Kaposi's sarcoma (KS) were amplified and sequenced. Four variants of HHV-8 were identified according to ORF 26 genotyping. Most of the samples were shown to be subtype A3, however, subtypes A, B3/C2/C2', and C3 (ORF 26 region) were also identified. The ORF 26 subtypes A and C3 of HHV-8 were only recovered from patients with KS while A3 was dominant in KS negative cases. The amplification of the hypervariable ORF K1 gene was successful only from 2 of the same 15 patients. Sequence analysis of the amplified ORF K1/VR1 regions identified subtype A3 from 2 patients with AIDS-associated KS. A novel ORF K1/VR1 variant belonging to subgroup A' was detected in a different sample in one of them. Amplification of the ORF K15, another representative locus for HHV-8 genotyping, was not successful from any of the peripheral blood samples. Unsuccessful amplification of the terminal K1 and K15 ORFs from peripheral blood samples suggests that KS biopsy specimens are needed for complete genotyping of HHV-8 strains from Hungary.

The geopathological significance of drinking water on the Great Hungarian Plain.

The author has stated that the teeth of the population of a village on the Great Hungarian Plain were much better in the last century than now concerning the quality of teeth. The water supply of the village has changed by the turn of the century, as the population began to drink the water of deep wells instead of that of the superficial ones. The relation between the decay of teeth and the change in the quality of drinking water could be detected on the basis of the chemical analysis of the water. The water of the last century contained much more fluoride than the water used at present. It must be pointed out that the change in the quality of drinking water was much more favourable from the point of view of epidemiology.

Molecular biological characterization of adenovirus DNA.

The agarose gel electrophoresis of DNA, the ethidium bromide fluorescence detection of DNA fragments and restriction endonucleases were discovered at the end of the '60s. The methodological progress enabled institutions equipped with less sophisticated technology to achieve also unique experimental and scientific results in the field of viral DNA research. The team working on virus DNA within the adenovirus research group has constructed several new restriction endonuclease maps of the genomes of human and animal adenoviruses; contributed to the methodology of the determination of specific endonuclease sites, and genome polarity; discovered new restriction endonucleases, adenovirus subtypes, new empty capsid, and genome subpopulations; participated in cooperations leading to novel, although hypothetical approaches in AIDS therapy, taxonomic definition of viruses, and evolutionary origins of adenovirus replication and encapsidation strategy.