[HCV genome variability in acute and chronic viral hepatitis C].

AIM: To evaluate HCV genome variability in acute and chronic phases of viral hepatitis C. MATERIAL AND METHODS: The study of heterogeneity of HCV in acute hepatitis C has detected genetic heterogeneity and variability of individual HCV population circulating in the blood. Significant genetic heterogeneity of HCV was observed in 1b, 2a and 3a genotypes. Variability of HCV did not depend on virus load. Genetic HCV structure changed significantly both in patients with manifest ALT deviations and in normal ALT, mean number of HCV genetic variants in these groups being the same. No significant correlations were found between virus concentration in the patient's blood, its variability and ALT values. Genetic heterogeneity of interferon-sensitive region of gene NS5A subtype 1b HCV was studied in blood of 16 patients with chronic hepatitis C resistant to interferon therapy. RESULTS: It is shown that genetic heterogeneity and variability of an individual HCV population circulating in blood serum can not be a prognostic criterion in assessment of variants of acute hepatitis C course. No mutations in ISDR region were found in 25% of 16 patients studied. 75% cases had 1-3 replacements of amino acid sequences, most frequent mutation was replacements in position 2218 (histidin/arginin). The above results are close to those obtained in Japanese and European populations. Results of ISDR sequence-analysis conducted before treatment may predict efficacy of interferon-alpha2 treatment in an individual patient in future. Large-scale trials are necessary for detection of mutations responsible for resistance to interferon-alpha2 in patients living in Russia.

[Synthesis and antigenic activity of peptides from the ORF3 protein sequence of hepatitis E virus]. / Sintez i antigennaia aktivnost' peptidov iz posledovatel'nosti belka ORF3 virusa gepatita E.

The hepatitis E virus (HEV) is an infection agent (detected recently) responsible for an enterically transmitted non-A, non-B hepatitis [1,2]. Hepatitis E is a big problem in many developing countries, including the Central Asian areas of the former Soviet Union [2]. By cloning followed by sequence analysis of the HEV genome, three open reading frames (ORF) have been identified, among them ORF3 encoding a protein containing 123 amino acid residues, the function of this protein being unknown. Recently [3], one of the immunodominant regions of ORF3 protein was revealed between the 91st and 123rd amino acid residue. The purpose of the present study was a more precise localization of epitopes in the C-terminal portion of HEV ORF3 protein by using synthetic peptides.

[Synthesis and immunological properties of the peptide fragment of the hepatitis B virus envelope preS-protein]. / Sintez i immunologicheskie svoistva peptidnogo fragmenta preS-oblasti belka obolochki virusa gepatita B.

We have synthesized the 24-41 fragment of the preS region of the hepatitis B (subtype ayw) envelope. The peptide was prepared by the solid phase synthesis on perfluoropoly-ethylene polymer grafted with polystyrene. The peptide chain was elongated from C-terminus using pentafluorophenyl- and p-nitrophenyl esters of Boc-amino acids. The peptide was cleaved off the solid phase by HBr in CF3COOH, purified by gel filtration, and, after conjugation with serum albumin (BSA), inoculated into mice. The resultant antibodies were shown to react with the peptide. The blood sera from patients with acute hepatitis B reacted with the peptide-BSA conjugates in the immunoenzymic solid phase assay.

[A comparative study of the diagnostic value of peptides, corresponding to various genomic sequences of the hepatitis delta virus]. / Sravnitel'noe issledovanie diagnosticheskoi tsennosti peptidov, sootvetstvuiushchikh razlichnym genomnym posledovatel'nostiam virusa gepatita del'ta.

A set of 5 peptides from the immunodominant region (65-80 aa) of delta-antigen was prepared by solid phase synthesis. Peptide 71-80 was synthesized in two variants with different amino acid residues in positions 73, 74, 76. Free peptides and their conjugates with bovine serum albumin were tested for antigenicity in ELISA. Correlation between the length of the peptide chain and its antigenic activity was noted. Peptides 65-80 and 69-80 demonstrated positive reaction with all sera from acute HDV patients. Both variants of peptide 71-80 reacted with 83% of positive sera. The smaller peptide 73-80 reacted strongly with one positive serum out of five used.

[Prevalence of hepatitis A markers in families of patients treated at home and in hospitals]. / Shirota rasprostraneniia merkerov gepatita A v sem'iakh bol'nykh pri lechenii ikh na domu i v statsionare.

The data on the time-course examinations of 119 family members of 59 adult patients with hepatitis A (HA) treated at home in Moscow and 170 relatives of 82 HA patients admitted to an infectious hospital are presented. The diagnosis for all the patients was verified by the detection in their blood of antibodies to hepatitis A virus (HAV) of the IgM class. Clinical methods of examination were used as well as determinations of anti-HAV of the IgG and IgM classes by enzyme immunoassay. When a case of HA was detected, the number of subjects susceptible to HAV in the families of the patients was insignificant (24-25%). The recorded cases of HA in adults in 60-70% were due to spread of the infection in the family; the sources of their infection were mostly children in whom this disease was frequently without jaundice and remained unrecognized. The incidence of infection of family members of HA patients treated at home or in hospitals was similar (10% and 9.5%, respectively) which proves the lack of epidemiological indications for mandatory hospitalization of such patients.

[Various physico-chemical properties of non-A, non-B hepatitis virus with a fecal-oral route of transmission and specific diagnosis of the illness]. / Nekotorye fiziko-khimicheskie svoistva virusa gepatita ni A ni B s fekal'no-orallnym mekhanizmom peredachi infektsii i spetsificheskaia diagnostika zabolevaniia.

Observations of 416 patients with and 112 convalescents after non-A-non-B virus hepatitis (HnAnB) with fecal-oral mechanism of transmission were carried out in 1984-1986. An enzyme immunoassay (EIA) was developed for the detection of HnAnB antigen in faeces. The rate of this antigen detection varied from 9.1% to 34.6% in different areas. The HnAnB antigen is present in two areas of cesium chloride density gradient: the maximum at 1.39-1.40 g/cm3 and another peak in the zone of 1.18-1.23 g/cm3. The method of radioimmunoprecipitation followed by SDS-polyacrylamide gel analysis showed the presence of several polypeptides of HnAnB virus. Most clearly, protein 30 K was documented, also polypeptides 91-94 K and 48-57 K were demonstrated. The resistance of nucleic acid of HnAnB virus to RNase was tested. The results indicate that the isolated NA is likely to be DNA. The above data suggest that the agent inducing HnAnB belongs to the group of parvoviruses.

[Analysis of the organization of immunodominant epitopes of hepatitis virus E orf2 and orf3 proteins using synthetic peptides]. / Analiz organizatsii immunodominantnykh épitopov belkov orf2 i orf3 virusa gepatita E s pomoshch'iu sinteticheskikh peptidov.

A set of synthetic peptides of different length derived from proteins encoded by open reading frames (ORF) 1, 2, and 3 of hepatitis E virus (HEV) genome was synthesized and tested for antigenicity in indirect enzyme immunoassay. Peptides 414-433 (ORF2 protein) and 99-119 (ORF3) specifically react with antiHEV-positive sera collected during outbreaks in the Central Asian areas of the former USSR. The antigenic structure of the Central Asian variants and the Burmese types of HEV was found similar.

[The etiological structure of acute viral hepatitis in Tadzhikistan in a period of decreased morbidity]. / Etiologicheskaia struktura ostrykh virusnykh gepatitov v Tadzhikistane v period snizheniia zabolevaemosti.

In the period of reduced incidence of viral hepatitis in Tajikistan, January-December, 1990, 1562 patients with acute viral hepatitis (AVH) were examined in the first days of the jaundice phase (928 children under 14 years and 634 adults) in Dushanbe. Markers of hepatitis A, B, and D (HBsAg, anti-HA IgM, anti-HBc IgM, anti-delta IgM) were determined by enzyme immunoassay. Hepatitis A occurred in 25.8% of the patients with AVH, mostly children of 1-6 years, HB in 22.8%, HD co- and superinfection) in 9.2%. In 42.1% of the patients who had no HA, HB, or HD markers in the blood, non-A, non-B hepatitis (mostly hepatitis E) was diagnosed, mainly in the age groups of 30-39 years (70.7%) and 15-29 years (59.2%). Thus, in Tajikistan hepatitis E occurs not only during outbreaks of this infection but also sporadically.

[IgM- and IgG-class antibodies to the hepatitis C virus]. / Antitela k virusu gepatita C klassov IgM iIgG.

The test system widely used currently for the determination of anti-HCV permits the detection of anti-HCV IgG alone. The data recently published by T. G. Wreghitt et al. confirm the probability of the presence of anti-HCV of both IgG and IgM classes in sera from hepatitis C patients. Anti-HCV IgM was detected by Ortho test with some modifications using an anti-M conjugate in the last stage of the experiment. Anti-HCV IgG were detected by regular Ortho test. A total of 46 patients with different forms of HCV infection and a control group were examined. According to the preliminary data, 18 patients were positive in the routine anti-HCV Ortho test. Among 18 anti-HCV-positive patients, nine had chronic HCV infection and the other 9 acute HCV infection. The distribution of IgM and IgG anti-HCV in the acute patients was as follows: 4 patients (44.5%) had approximately equal titres of IgG and IgM, 3 (33.5%) had predominantly IgG, 2 (22.2%) mainly IgM. A similar pattern was observed in the group with chronic HCV infection. Thus, 5 subjects (55.6%) showed approximately equal ratio of IgM and IgG anti-HCV, 2 (22.2%) had mostly IgM and the rest 2 mainly IgG. No anti-HCV in the control group was found. The control group consisted of 18 patients with chronic liver diseases without markers of HBV or HDV infection, 3 with HAV infection, 2 with HBV infection and 5 healthy subjects. The specificity of anti-HCV IgM test was confirmed by Chiron Western blot analysis using the same modification.(ABSTRACT TRUNCATED AT 250 WORDS)

[The prevalence of the markers of viral hepatitis B and delta among the population in regions differing in the level of morbidity]. / Raspprostranenie markerov virusnykh gepatitov B i del'ta sredi naseleniia regionov, kontrastnykh po urovniu zabolevaemosti.

From two regions differing by the levels of incidence of hepatitis B, 2019 blood serum specimens from normal population were examined for markers of HBV infection. In Moscow, among 1040 samples examined HBsAg was found in 2.0%, anti-HBs in 10.0%, anti-HBc in the absence of HBsAg and anti-HBs in 4.5%. In the Osh Province of Kirgizstan, among 979 subjects examined the same markers were found in 10.3%, 22.4%, and 14.0%, respectively. In this area, HBsAg was detected most frequently among infants (14.9% in infants under 1 year), in whom HBs-antigenemia was combined with the presence of HBeAg in 54.5% and with anti-HBc-IgM in 69.2%. Antibody to delta antigen (anti-delta) was found in 24 (25.8%) out of 93 HBsAg-positive subjects in the Osh Province but in none of 21 subjects with HBs-antigenemia in Moscow.

[The production of anti-HBc IgM antibodies by human lymphocytes transformed by the Epstein-Barr virus]. / Produktsiia anti-HBc IgM-antitel limfotsitami cheloveka, transformirovannymi virusom Epshteina--Barr.

In transformation by Epstein-Barr virus of lymphocytes derived from patients with acute hepatitis B, continuous cell cultures were obtained which produced anti-HBc IgM antibodies. These cell lines underwent from 50 to 150 passages. The level of the specific immunoglobulin production was shown to have a trend to decline; however, after cloning the antibody production became more stable. The antibodies produced by the cloned lymphocyte culture could not neutralize the antigen (unlike polyclonal antibodies) which indicated a high efficacy of cloning. When the solid phase was sensitized with produced immunoglobulins, specific activity of binding of HBc antigen detectable by anti-HBc conjugate with horseradish peroxidase was demonstrated. The study of sedimentation properties of antibodies produced by transformed lymphocytes showed their sedimentation constant to be 19S.

[The incidence of detecting hepatitis C viral antibodies in different age groups of the population in northeastern Ukraine]. / Chastota obnaruzheniia antitel k virusu gepatita C v razlichnykh vozrastnykh gruppakh naseleniia severo-vostochnoi Ukrainy.

Examinations for the presence of antibodies to hepatitis C virus (anti-C100-3) were carried out in 1143 human subjects, residents of the town of Sumy, Ukraine (681 blood donors, 176 school children, 286 subjects hospitalized for abortions and for treatment of acute traumas). Anti-HCV were determined by enzyme immunoassay using a commercial test system "Abbott" of the first generation, with confirmation of the positive findings by neutralization test. Anti-HCV were found in 2.3% free-giving blood donors and in 1.2% career donors, and in 0.8% of schoolchildren aged from 6 to 14 years. In older age groups anti-HCV findings were more frequent (in subjects of 40-49 years 4.0%, 50-59 years 5.26%, 60 years and over 6.25%).

[Characteristics of viral hepatitis non-A, non-B with a fecal-oral mechanism of transmission of the infection in southern Uzbekistan]. / Kharakteristika virusnogo gepatita ni A ni B s fekal'no-oral'nym mekhanizmom peredachi infektsii v iuzhnom Uzbekistane.

An increase in the incidence of viral hepatitis was observed in the autumn of 1986 in one of the districts of southern Uzbekistan. The clinico-epidemiological data and highly sensitive tests for hepatitis A (HA) and B (HB) markers showed the absence of HB infection in most of the patients and prevalence of HA among children. In 68.7% of the patients, non-A-non-B hepatitis with fecal-oral mechanism of transmission of the infection was diagnosed. The bulk of HnAnB patients consisted of adults of 20-29 years. Pregnant women experience severe forms of the disease. Solid-phase enzyme-immunoassays and immune electron microscopy revealed in one fecal specimen from patients the antigen of non-A-non-B hepatitis serologically identical to that causing a rise in the incidence in northern Turkmenia in 1984-1985.

[Prevalence of genotypes of the hepatitis C virus, circulating in northwestern and central parts of Russia]. / Rasprostranenie genotipov virusa gepatita C, tsirkuliruiushchikh na territoriiakh severo-zapadno i i tsentral;no i chaste i Rossii.

Testing of 90 sera for antibodies to hepatitis C virus (HCV) by genotyping methods resulted in determination of the genotype in 83 cases: 47 cases with 1b genotype, 27 with 1a, 7 with 3a, 1 with 2a, and 1 with 2b genotype. Hence, preliminary data indicate the predominance of HCV genotype 1b among patients with hepatitis C in these regions of Russia.

[Regularities in the spread of hepatitis C virus and its genotypes in Russian and countries within the former USSR]. / Zakonomernosti rasprostraneniia virusa gepatita C i ego genotipov v Rossii i stranakh SNG.

The incidence of markers of hepatitis C virus (HCV in the blood of 4216 normal subjects living in the European Russia (Northern, North-Western, Central, Central Chernozem, Volga-Vyatka, Volga, and North Caucasian regions), in the Urals, in Siberia (Eastern Siberian region), in the Far East, and in Monogolia is assessed. The incidence of antibodies to HCV varied from 0.7% in the Central region to 3.8% in the Central Chernozem and 10.7% in Mongolia. HCV genotyping (identification of 1a, 1b, 2a, 2b, and 3a genotypes) was performed using 469 RNA of HCV-positive sera of donors and patients collected in Russia, Moldova, Turkmenistan, and Mongolia. The 1b genotype predominated everywhere (68.9%), its incidence being the highest in Moldova (96%). Unclassifiable variants of HCV were found in 28 (6%) of sera. The regularities of HCV genotypes circulation in the European Russia were the same as in other European countries, whereas their prevalence in Eastern Russia was rather like that in China or Japan. The prevalence of genotypes did not depend on the clinical manifestations of diseases caused by HCV.

[Morbidity and mortality in acute viral hepatitides in a hyperendemic region. The role of viral hepatitis E]. / Zabolevaemost' i smertnost' pri ostrykh virusnykh gepatitakh v giperéndemichnom regione. Rol' virusnogo gepatita E.

Official annual statistical data on morbidity in acute viral hepatitides (AVH), including the number of lethal cases, for 1985-1995 were analyzed. Mortality rates per 100,000 of the population at the period of 11 years were calculated for different age groups, sex and the place of residence. 396 and 99 patients were examined for the presence of serological markers of hepatitides A, B and E, respectively, at the periods of epidemic rises in morbidity and satisfactory epidemic situation. In the course of 11 years AVH caused the death of 22,405 persons. In 1985-1987 the average mortality level (ML) reached 12.3-17.8 per 100,000 of the population (with morbidity being 1,200-1,400 and was essentially higher among the rural population in comparison with the urban population. During these years the highest ML, was registered among children aged 0-2 years (190-50 per 100,000) and, among adults, mainly among women aged 20-29 years (21.4-19.6 per 100,000). During the years when the epidemic of AVH was absent, ML among these groups was essentially lower: 40-20 among children aged 0-2 years and 4-5 among women aged 20-29 years. In 1987 in the Fergana Valley hepatitis E was detected in 72.2% of all examined patients, and in the southern areas of the country in 68.7%. A sharp rise on mortality among women of the productive age at the period of the epidemic rise of AVH morbidity in the endemic region indicated that this epidemic was linked with hepatitis E. High ML among young children may be indicative of a highly unfavorable course of hepatitis E in the group of infants, which had never been registered before. This newly established regularity may be used for the retrospective diagnostics of the outbreak of hepatitis E.

[The characteristics of an outbreak of non-A, non-B hepatitis with a fecal-oral mechanism of the transmission of the causative agent in Osh Province, the Kirghiz SSR]. / Kharakteristika vspyshki gepatitia ni A ni B s fekal'no-oral'nym mekhanizmom peredachi vozbuditelia v Oshskoi oblasti Kirgizskoi SSR.

The results of epidemiological, clinical and laboratory studies revealed that the sharp rise of morbidity in viral hepatitides in Osh Province, the Kirghiz SSR in autumn 1987 was caused by hepatitis non A, non B virus with fecal-oral transmission. At this period the results of the enzyme immunoassay showed the absence of the markers of hepatitides A, B and Delta in 72.2% of viral hepatitis patients. Hepatitis non A, non B occurred only in 2.4% of viral hepatitis patients of preschool age (of these, 83.3% had hepatitis A) and was diagnosed in autumn 1987 in 50% of the patients aged 7-14 years and in 97.4% of the patients aged 15-29 years (in the latter age group 95-98% of the patients had IgG to hepatitis A virus in their blood). The appearance of the outbreak of the above-mentioned infection in Kirghizia is linked with the water route of the transmission of the infective agent. The epidemiological and clinical signs, characteristic of fecal-oral hepatitis non A, non B in Kirghizia, were not different from those registered earlier in other republics of the Central Asia and could be used for the identification of this infection.

[The spread of hepatitis C markers among the population of regions of Russia and Central Asia]. / Rasprostranenie markerov gepatita C sredi naseleniia nekotorykh regionov Rossii i Srednei Azii.

A total of 2,217 blood serum samples taken from the population of some regions of the former USSR were studied for the presence of antibodies to hepatitis C virus (HCV) with the use of a commercial enzyme immunoassay system (obtained from Ortho, Japan). Perceptible changes in the detection rate of these antibodies throughout the territory of the former USSR were established. Most frequently anti-HCV antibodies were detected among the population of the republics of Central Asia (5.3-2.9%), as well as Tuva and Yakutia (2.5-3.0%), their lowest detection rate was in Moscow (1.3%). In Moscow the results of the assay in adults were positive twice as frequently as in children, and in donors with an elevated level of alanine-transferase activity the presence of these antibodies was confirmed in 8.9% of cases. The frequency of the confirmation of the presence of anti-HCV antibodies by recombinant immunoblotting was 70.7% in all positive sera and 100% in the sera from a group of donors with an elevated enzyme level. In many respects coincidence between the regularities in the spread of viral hepatitic C and hepatitides B, D was noted.

[Characteristics of the course and outcome of viral hepatitis A in patients treated at home]. / Kharakteristika techeniia i iskhodov virusnogo gepatita A pri lechenii bol'nykh na domu.

As many as 40 adult patients aged 15 to 40 years with virus hepatitis A treated at home were followed up since October 1987 to March 1988 in 6 districts of Moscow. There were 23 women and 17 men. The control group comprised 101 adult patients (46 women and 51 men) admitted to the Clinical Infectious Hospital No. 1. The diagnosis of virus hepatitis A was made on the basis of the clinico-epidemiologic and biochemical data, supported by detection of specific IgM antibodies to virus hepatitis A. Staying in bed, sparing diet and abundant drinking were indicated to all the patients. The patients who reported for work were followed up for 6 months. The mean duration of disability constituted 34.1 +/- 1.69 days, if the patients were treated at home and 38.6 +/- 1.26 days, provided they received hospital treatment. The number of lingering cases was 4 (10%) during the treatment at home and 11 (10.9%) at hospital treatment. The data obtained attest to the possibility of treating adult patients with virus hepatitis A at home.

[The clinical picture and outcome of acute delta infection]. / Klinika i iskhody ostrogo del'ta-gepatita.

A total of 60 patients were observed with acute hepatitis delta defined at examination of 200 adult subjects with acute HBsAg-positive hepatitis. Moderate and severe forms of the disease occurred more frequently, lethality reached 13.3%. An unfavorable prognosis was related to a short-term prejaundice period with intoxication (high body temperature, head ache, vertigo, recurrent vomiting, IgM antibodies presence in the serum at the height of the disease). No patients achieved cure within 9 months following the discharge from hospital. The process acquired the form of long-term convalescence with transformation into chronic hepatitis delta.