[ASSOCIATION OF POLYMORPHISM GENES OF SURFACTANT PROTEINS IN PATIENTS WITH INFLUENZA].

AIM: Evaluate role of gene polymorphisms of surfactant proteins in susceptibility and severity of influenza infection course in representatives of Moscow population. MATERIALS AND METHODS; 320 influenza patients, infected with various influenza virus strains, and 115 healthy individuals (control group),, were included into the study. Human DNA samples genotyping for determination of SFTPA2 gene rs1965708 and rs1059046, SFTPB gene rs1130866 polymorphisms was carried out using a modified method of "adjacent samples". RESULTS: Most of the individuals of the control group and influenza patients are carries of alleles and genotypes rs1965708 and rs1059046 of SFTPA2 gene, rs1130866 of SFTPB gene, that have, based on scientific literature data, shown association with severe course of influenza A(H1N1) pdm09 and other inflammatory diseases of the respiratory tract. Generally, significant differences in frequency of occurrence of unfavorable genotypes CC rs1965708, AA rs1059046 of SFTPA2 gene and CC rs1130866 of SFTPB gene in influenza patients in comparison with individuals of the control group were not detected, that gives evidence on a high (from 19 to 51%) prevalence of these genotypes in the studied population. Allele C and genotype CC rs1965708 of SFTPA2 gene, allele A and genotype AA rs1059046 of SFTPA2 gene, allele C and genotype CC rs1130866 of SFTPB gene did not shown an association with severe course of A(H1N1) pdm09 influenza. The following pathology registered in most (88%) of the patients with severe course of influenza A (H1N1)pdm09: diseases of cardiovascilar (44%), endocrine (36%) and respiratory (12%) systems. CONCLUSION: Because in most of the deceased patients due to severe course of A (H1N1)pdm09 influenza, diseases of cardiovascular, respiratory and endocrine system were detected, and an association of unfavorable disease outcome with the studied genetic markers was not detected, dominating risk factor of development of severe course and lethal outcome for A(H1N1)pdm09 influenza in the studied cohort was comorbidity.

[The problem of viral hepatitis C in the Russian Federation].

The incidence of chronic viral hepatitides (CVH) has increased 2.2-fold in the Russian Federation over the past decade. This increase is mainly determined by an almost threefold rise in the incidence of chronic hepatitis C (CHC): from 12.9 in 1999 to 39.1 per 100,000 population in 2012. The calculated data of hepatitis C burden in the Russian Federation show that in 2010 the total medical and social losses and expenses associated with hepatitis C and its implications were 48.47 billion rubles or 0.108% of the gross domestic product, the direct medical costs were 17.1 billion (35.28%) rubles, GDP losses were 26.05 billion (53.75%) rubles, and the disability payments were 5.32 billion (10.97%) rubles. The patients (mean age 45 years) with liver cirrhosis (LC) were 15.2% in the structure of the CHC patients (mean age 37 years) admitted to Moscow infectious diseases hospitals in 2010. Analysis of the regional registers of the Russian Federation, the proportion of patients with LC among those with CHC was 18%. The existing forms for recording morbidity and mortality from poor CHC outcomes cannot significantly estimate the true disease stage distribution of patients and hepatitis C-associated disability and mortality rates. In this connection, it is necessary to introduce a federal register and to change recording forms for patients with viral hepatitides. Standard interferon, pegylated interferon alpha 2a and pegylated alpha 2b, and the HCV protease inhibitors telaprevir, boceprevir, and simeprevir have been registered for the treatment of hepatitis C in the Russian Federation.

[Features of proliferative response of T-cells on hepatitis c virus antigens in healthy individuals contacting with this virus].

AIM: Evaluation of CD3+ and CD3-/CD56+ proliferative response on hepatitis C virus antigens in healthy medical workers. MATERIALS AND METHODS: The study included 15 medical workers with length of service of 2 and more years without common risk factors (blood and blood products transfusion, abdominal operations, invasive procedures, use of intravenous drugs). Control group consisted of 9 healthy individuals without risk factors. Peripheral mononuclears were isolated from blood and then incubated 72 hours in the presence of mitogen/PHA, Core+NS4 1b genotype HCV, NS4 HCV2a+3a genotypes or medium. Proliferative activity was registered by the presence of cell division marker ki67 by using FACS. RESULTS: The initial immunogram of medical workers differed from the control group by a significantly lower quantity of CD3+ lymphocytes, in particular CD3+/CD8+ population. Incubation with PHAresulted in a decrease of quantity of CD3+/ ki67+, CD4+/ki67+ and CD3-/CD56+/ki67+ in the medical workers group. Cultivation with HCV antigens resulted in a significant decrease of Treg (CD3+/CD25high/FoxP3+) and activated T-lymphocytes population in the case of stimulation by Core and NS4 1b genotype antigens. Analysis of cell response on virus antigens based on proliferative activity index detected significant differences only for CD8+/ki67+. Stimulation by Core and NS4 1b genotype antigens resulted in an increase of quantity of these cells whereas in the case of NS4 2a+3a genotypes their decrease was observed. CONCLUSION: The described changes may reflect exhaustion of cell reactivity due to extra antigen load in the group of medical workers, while differentiated immunologic shifts on hepatitis C viruses of various genotypes are noted.

[Immunogenetic factors of interaction of hepatitis C virus and human and possibilities of development of therapeutic tactic].

Various macroorganism factors influence spontaneous and therapy induced elimination of hepatitis C virus. Standard therapy is application of a combination of pegylated interfernos and ribavirin. However such tactics results in attainment of a resistant virological response in approximately half the cases during infection by genotype 1 virus. The future of viral hepatitis C therapy lies most probably in the sphere of application of specific antiviral preparations such as protease and/or polymerase inhibitors as an addition to standard therapy. The aim of this review was to describe mechanisms of resistance to therapy in light of reaction of innate and adaptive immune response as well as an attempt to determine factors capable of prognosing response to therapy

[The burden of viral hepatitis in the Russian Federation requires reducing its progression for the long term, including chronic hepatitis C)].

AIM: To evaluate the impact of using the innovative antiviral drugs in patients with chronic hepatitis C (CHC) on the disease burden in the long term. MATERIALS AND METHODS: A program was developed to increase the availability of innovative antiviral therapy (AVT) in patients with CHC covering the period from 2013 to 2020. To evaluate the impact of the program on the burden of hepatitis C (HC) in the Russian Federation, a Markov model of the disease progression was developed till 2030. Calculations were made in medical and social perspectives. RESULTS: The implementation of the program will allow to decrease the medical costs by RBL 2 billion and budget costs, including disability payments, by RBL 3.8 billion by 2030. During this period, in GDP losses will decrease by RBL 1.1 trillion and the total burden of HC taking into account the cost of the program by RBL 1 trillion. CONCLUSION: For the first time it was shown that the expanding coverage of CHC patients with innovative AVT will allow to reduce the costs of health and social services.

[Immunogenetic predictors of a rapid virologic response to antiviral therapy in patients with chronic hepatitis C].

AIM: To determine the correlation between interleukin 28B (IL28B) gene polymorphism in patients with chronic hepatitis C (CHC), the presence or absence a rapid virologic response to antiviral therapy, and a number of immunological characteristics as a basis for a personalized approach to treating the patients. SUBJECTS AND METHODS: Seventeen CHC patients infected with hepatitis C virus genotype 1b were examined and underwent genetic testing for IL28B gene polymorphism for rs12979860 (CC, CT or TT genotypes) and rs8099917 (TT, TG or GG genotypes) using the modified method of adjacent samples, which revealed single nucleotide substitutions in the genes. Their immunological parameters were identified by a flow cytometry technique by taking into account whether a rapid virologic response had been achieved. RESULTS: The key phenomena of a rapid virologic response in the representatives of different IL28B genotypes are the nonspecific proliferative activity of blood natural killer cells before treatment, as well as the count of regulatory T cells before and 4 weeks after therapy start. CONCLUSION: To predict the efficiency of antiviral therapy for CHC, it is desirable to supplement genetic studies with immunological data.

[Long-term entecavir efficacy retention in the treatment of chronic hepatitis B with an outcome to hepatic cirrhosis caused by its resistant virus strain].

The paper gives concise data on the efficacy of entecavir in the treatment of chronic hepatitis B caused by its resistant virus strain and describes a relevant clinical case.

[The etiological pattern of diseases in pregnant women with enhanced blood ALT and AST activities, admitted to the obstetric unit of infectious disease hospital].

AIM: to define a role of hepatotropic (HAV, HBV, HCV, and HDV) and opportunistic hepatotropic (HGV, CMV, EBV, HHV types 1, 2, and 6) viruses in the etiological pattern of diseases accompanied by enhanced blood AlAT and AsA T activities in pregnant women. SUBJECTS AND METHODS: Two hundred and eleven pregnant women, including 123 patients with chronic viral hepatitis, 74 with enhanced blood AlAT activity and no markers of viral hepatitis (EAlA T-NMVH), and 14 with acute viral hepatitis were examined. RESULTS: Most pregnant women with chronic HBV and HCV infections were found to have HBV DNA and HCV RNA in the blood in the presence of normal and enhanced activities of transaminases. In the EAlAT-NMVH group, there was none of the opportunistic hepatotropic viruses in more than 7% of cases. No genetic material of HAV, HBV, HCV, HDV, HGV, CMV, EBV, HHV types 1, 2, and 6 was found in the blood of all 10 patients with hepatitis of unspecified etiology. CONCLUSION: In the absence of serologic data supporting the presence of infectious pathology, blood testing using the polymerase chain reaction is of low informative value in detecting opportunistic hepatotropic viruses in pregnant women with hepatitis of unspecified etiology. However, by keeping in mind that the spectrum of opportunistic hepatotropic viruses is not confined to those included in this study, it is expedient to examine additionally pregnant women with enhanced blood AlAT and AsAT activity in order to identify TTV, B19V, HHV-8, SEN and NV-F in the blood.

[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.

[Acute hepatitis of unspecified etiology: etiological structure and clinical-laboratory characteristics].

The straight line nucleic acids detection method of viruses and wide spectrum of virus antigens immunodiagnostics in acute hepatitis of unknown etiology patients has allowed verifying the diagnosis at 19% cases (a viral hepatitis A, C or E). Results of research do not allow to consider hepatotropic viruses HGV, TTV, PV B19, EBV, CMV, HHV 1, 2, 6 and 8 type, NV-F as etiological agents at the majority of patients of investigated group, and the data of the anamnesis and a clinical and laboratory picture of a current of disease does not allow to exclude at 29.4% of patients a drug-induced hepatitis. Despite detailed molecular-biological and immunological inspection of patients, at 37.9% of acute hepatitis of unknown etiology patients it was not possible to establish a connection with hepatitis and defined etiological factor (the infectious agent).

[Role of host in spontaneous recovery from viral hepatitis C: the body's condition at the moment of infection].

The condition of the host at the moment of infection is that its immune competence largely determines the efficiency, kinetics, and profile (Thl/Th2) of a further specific immunity response and, accordingly, the outcome of penetration of hepatitis C virus (HCV) into the body and subsequent acute infection (if it occurs). The parameters determining immune competence may include age, traumatizing exposures (operations, burns, wounds, and fractures), immunosuppressive therapy, stresses, con-infections, and alcohol use. The highest rates of spontaneous convalescence from HCV infection are observed in children and adolescents. Other human conditions are much shorter, transient; their impact is difficult to determine in the retrospective review and therefore it has not been adequately studied. Previous operations, posttransplantation immune suppression, immune modulation after blood transfusion, alcohol-induced immune imbalance, drug and narcotic intoxication are poor predictors. Immunosuppression and immune imbalance caused by viral and parasitic infections are observed among the host's temporary conditions affecting the outcome of HCV infection. The authors have analyzed the sequels of superinfections in patients with chronic hepatitis C, other hepatotropic viruses and the common liver fluke Schistosoma mansoni. The interesting therapeutic activities against HCV and parasitic infection (contamination with Echinococcus granulosus in particular), which are shown in the treatment of co-infection patients with alpha-interferon preparations that ensure normalization of immune deficiency caused by each of the infections and their increased combination. A deeper insight into the correlation between the condition of the host and its ability to eliminate the virus may be one more step on the road to the prevention of the infection and to the designing an effective vaccine against HCV.

[Blood levels of HbsAg in patients with different variants of chronic HBV-infection].

AIM: To estimate HbsAg in patients with different variants of chronic HBV infection. MATERIAL AND METHODS: Assay of HbsAg (IU/ml) in blood serum was made in 156 patients with chronic HBV infection (70 males and 86 females, age 19 to 78 years) using the test-system HbsAg Architect Lot 59665LF00 (Abbott) on the automatic analyzer Architect with construction of 4-parameter logistic curve. RESULTS: There are significant differences in the levels of HbsAg depending on the course of chronic HBV-infection: inactive carriers of HBV (12,884.14 +/- 5,512.26 IU/ml) had much lower blood levels of HbsAg than patients with HbeAg-negative (66,992.28 +/- 25,908.74 IU/ml) and HbeAg-positive chronic VHB (135,039.3 +/- 48,127.06 IU/ml) patients with chronic mixed hepatitis (82,783.12 +/- 21,001.34 IU/ml) and cirrhosis of HBV-etiology (67,477.86 +/- 24,081.9 IU/ml). No significant differences were found between two subgroups of pregnant women with or without viremia by HbsAg concentration in the blood. Maximal mean content of blood HbsAg was registered in patients with HbeAg-positive chronic hepatitis B. CONCLUSION: Significant differences in blood serum levels of HbsAg exist between patients with chronic viral hepatitis B and inactive carriers of HBV.

[Diagnostic significance of type IV collagen and hyaluronic acid in the serum of patients with chronic hepatitis C for staging hepatic fibrosis].

AIM: To evaluate diagnostic value of serologic fibrosis markers (hyaluronic acid--HA and type IV collagen C-IV) in patients with chronic hepatitis C (CHC) and hepatic cirrhosis (HC). MATERIAL AND METHODS: HA and C-IV were measured in 88 CHC patients with fibrosis stage 1 (n = 63) and 3 (n = 25), 13 patients with acute hepatitis C (AHC), 28 patients with hepatic cirrhosis (HC), 19 patients with pulmonary fibrosis (PF). The control group consisted of 32 healthy subjects. RESULTS: HA concentrations in the serum of CHC patients with mild to severe inflammation and fibrosis (F1 and F3) were normal (100 ng/ml). For HC diagnosis, HA test proved highly sensitive and specific (in HA 100 ng/ml sensitivity was 100%, specificity 84.6%), but this method cannot stage hepatic fibrosis. HA test was inferior to C-IV test. A mean C-IV concentration in the serum of CHC patients at the stage of marked fibrosis (F3) is significantly higher than in F1, in HC (A) patients higher than in patients with CHC F3. CONCLUSION: It is shown than concentration of C-IV above 196 ng/ml can differentiate fibrosis stage 1 from stage 3 with specificity 58.7 and sensitivity 88%.

[Immune status of patients with different outcomes of acute hepatitis C].

AIM: To ascertain parameters of immune response significant for prognosis of the disease outcome in patients with acute hepatitis C (AHC). MATERIAL AND METHODS: Seventy two examinees were divided into three groups: 30 patients of group 1 had AHC; 29 patients of group 2 had chronic hepatitis C (CHC) with the disease history 3-10 years; 13 AHC convalescents who had the disease 3-10 years ago. The control group consisted of 10 healthy subjects. The following parameters of immune status were studied: CD3+, CD4+, CD8+, CD16+, CD20+, HLA-DR+CD3+, CD95+, O-lymphocytes. RESULTS: By clinicobiochemical picture and the results of 12-month follow-up RT-PCR investigations of blood HCV RNA, AHC patients were divided into two groups: in 18 (72%) of 25 patients AHC became chronic, 7 (28%) patients achieved convalescence. The proportion and absolute number of subpopulations of lymphocytes CD8+, CD20+, CD16+ as well as CD4/CD8 in patients with AHC with different outcomes did not differ from the controls. AHC convalescents and patients with AHC transformation into chronic hepatitis had in the acute period of the disease much higher absolute number of subpopulations of the lymphocytes CD3+, CD4+, HLA-DR+CD3+ vs the controls. In patients with chronic hepatitis C with elevated levels of AlAT, 3-10 year follow-up registered significantly higher absolute amount of CD8+, CD3+, CD4+, HLA-DR+CD3+ vs the controls. Only patients with AHC which later became chronic had for 6 months of the disease significantly elevated absolute number of O-lymphocytes vs the controls. CONCLUSION: A high absolute count of O-lymphocytes in AHC patients may indicate probable development of chronic hepatitis C.

[Detection of type specific antibodies to hepatitis C virus NS4 protein in hepatitis C patients by enzyme immunoassay]. / Vyiavlenie tipospetsificheskikh anti-HCNS4-antitel u bol'nykh gepatitom C metodom immunofermentnogo analiza.

A multi-enzyme immmune-assay test system was designed for serotyping of genotypes hepatitis C virus (HCV) and a method of such typing of the serum of patients with hepatitis C was worked out. The above test-system was worked out on the basis of a study of 10 type-specific peptides modeling different fragments from NS4-protein variable region of HCV. The designed test system was evaluated by using a set of 42 serum samples obtained at random from patients with chronic hepatitis C, which had been preliminarily genotyped by polymerase chain reaction. The serotyping makes it possible to identify the type-specific antibodies in the blood sera of patients, including those cases when viremia was absent. Differences in the circulation of HCV in Moscow (Russia) and Vitebsk (Byelorussia) were established by using the designed test-system.

[Seroconversion to various hepatitis C virus antigens in patients with hepatitis C from various sources]. / Issledovanie serokonversii k razlichnym antigenam virusa gepatita C s razlichnymi iskhodami.

A total of 136 patients with acute icteric hepatitis C, including patients with known outcome, were examined. Therefore, 46 serological samples, obtained from 13 patients with subsequent remission, and 63 samples, obtained from 13 patients, who subsequently developed the chronic disease stage, were analyzed. The serum of known outcome patients were examined, by using the immune-enzyme analysis method, to the antibodies of both class IgG, and IgM. Differences in the dynamics of the immune humoral response were established with due regard for a pathological course process, including the acute disease stage. The obtained results are interesting because of their prognostic values.

[Spectrum of antibodies to HCV antigens in patients with different clinical patterns of chronic HCV infection]. / Spektr antitel k razlichnym antigenam HCV pri raznykh variantakh techeniia khronicheskoi HCV-infektsii.

Correlations between the spectra of antibodies to HCV proteins represented by various antigenic determinants and clinical variants of chronic HCV infection were studied. Synthetic peptides core-16, NS4-20, and NS5-23 simulating the immunodominant regions of the core, NS4 and NS5 proteins and recombinant proteins core-114 and NS4-86 were used as antigens. The results indicate that if the serum of an HCV patients contains no IgG to both antigenic determinants of NS4 or to NS5 in combination with any core antigenic determinant, a clinical and biochemical remission is highly probable. Chronic hepatitis C is characterized by the presence of IgG in high titers to both antigenic determinants of NS4 protein, particularly in combination with anti-NS5 IgG in low titers or none at all, or high titers of anti-core-16 IgG in combination with high titers of anti-NS4-20 IgG.

[Diagnostic significance of antibody detection to various hepatitis C virus antigens in patients with acute and chronic HCV-infection]. / Diagnosticheskaia znachimost' opredeleniia antitel k razlichnym antigenam virusa gepatita C u patsientov s ostroi i khronicheskoi HCV-infektsiei.

AIM: To examine diagnostic value of antibodies to various HCV antigens in patients with acute and chronic HCV-infection. MATERIAL AND METHODS: Enzyme immunoassay has tested blood sera from 136 patients with icteric acute hepatitis C (AHC) and 45 patients with chronic HCV infection for IgG antibodies to antigens of proteins core, NS4, NS5, HCV. Synthetic peptides core-16, NS4-20, NS5-23 were used as antigens. RESULTS: Patients with icteric AHC had IgG antibodies to antigens of both structural protein core and non-structural proteins NS4, NS5 of HCV as early as the first 10 days of jaundice. Occurrence of anti-core and anti-NS4 increases with the disease duration. Incidence of anti-NS4 correlated with duration of previous intravenous drug addiction. In patients with AHC early in the icteric period anti-core, anti-NS4, anti-NS5 were present less frequently than in patients with chronic HCV infection having elevated levels of AlAT. Significant differences were found neither with the group with normal AlAt nor in the spectrum of the detected antibodies between patients with acute and chronic HCV infection. CONCLUSION: Despite different frequency of anti-core, anti-NS4, anti-NS5 detection in patients with icteric AHC and patients with chronic HCV-infection and high AlAT, their high incidence rate in this or that group and absence of differences by the spectrum of the studied antibodies do not allow the fact of their detection to be a diagnostic marker differentiating acute HCV-infection with chronic one.