Estimation of efficiency of solvent-detergent method for virus inactivation in the technology of immunoglobulin production on the model of duck hepatitis B virus.

The virucidal action of solvent tributyl phosphate and detergent sodium cholate used in the production of immunoglobulin for inactivation of viruses with lipid envelope was studied on the model of duck hepatitis B virus. PCR analysis revealed no significant decrease in duck hepatitis B virus DNA concentrations after treatment with solvent/detergent. At the same time, in vivo experiments showed that treatment of duck hepatitis B virus with tributyl phosphate (concentration >0.15%) and sodium cholate (concentration >0.1%) at 37°C for 6 h or longer completely inactivated this model virus added to immunoglobulin solution in concentration 5 log ID50. Duck hepatitis B virus appears to be one of the most acceptable model viruses for validation of virus inactivating technologies in manufacturing human plasma preparations.

[Specific humoral immunity after single immunization with mumps vaccine: data of a 3-year follow-up].

The level and spectrum of humoral specific immunity were studied in 60 volunteers immunized with Russian mumps vaccine. Specific IgG levels were measured by enzyme immunoassay (EIA) and neutralization test using the Leningrad-3 (L-3) mumps virus (MV) vaccine strain and 5 heterologous MV strains of various genotypes (A, B, C, D, and H). The maximum functional activity of antibodies was recorded at an average of 18 months postvaccination. Within 3 years after vaccination, starting at 6 months, specific IgG neutralized all 6 MV strains having varying activity in relation to the genotype. Neutralizing titers (NT) against the L-3 strain were 1.3-1.7-fold higher than those against heterologous MV strains throughout the follow-up. Despite a tendency towards lower specific IgG levels, within 3 years postvaccination, EIA IgG titers remained to be 2.5 -log, L-3 strain HT were -log, or more, and the titers against 5 heterologous MV strains were 2 -log2 or more in all the volunteers.

[Functional state of specific immunity in children and teenagers vaccinated against mumps].

The functional state of immunity was evaluated from the avidity index (AI) of specific antibodies (IgG) and the level and spectrum of their neutralizing activity. The study recruited 200 subjects immunized with Russian vaccine against mumps according to the mandatory scheme. A group of vaccinees with a low AI of specific IgG was identified mainly among old children and teenagers. The vaccinees with a low AI had a significantly lower protective immunity (as shown from the level and spectrum of serum neutralizing activity) than those with a high AI. The vacinees with no humoral, incomplete, or complete postvaccination immunity, but with a low AI of specific IgG, can constitute a population stratum that preserves sensitivity to wild-type mumps viruses and serves as a favorable medium for their circulation.

[The immunogenic and protective properties of inactivated and live candidate vaccines against highly pathogenic avian influenza H5N1 virus].

The present study in BALB/c mice was conducted to compare immunogenicity and protective efficacy of several candidate vaccines based on homologous and heterologous strains after challenge with the highly pathogenic avian influenza strain A/Chicken/Kurgan/3/2005. The experimental vaccine composed of an inactivated split A/Vietnam/1194/2004 (H5N1) strain and a plant derived adjuvant has demonstrated better immunogenic properties versus the variant of the vaccine with aluminum hydroxide. Interestingly, the heterosubtypic H1N1 live attenuated vaccine candidate administered intranasally protected 93% of the subject against their challenge with HPIV HSN1.

[Virus strain specific serum neutralizing antibodies in children and adolescents immunized with a Russian mumps vaccine].

Postvaccination immunity was studied in the children and teenagers without a history of clinical mumps infection, who had been immunized with the Leningrad-3 mumps vaccine. The level of specific lgG in ELISA and that and spectrum of their neutralizing activity against a vaccine strain and three heterologous mumps virus (MV) strains (genotypes A, C, and H) were measured. The investigation included 151 sera from the vaccinees aged 3 to 17 years, possessing the detectable specific IgG titers in ELISA and the detectable neutralizing titers against the vaccine strain. 97.4% of the vaccinees had neutralizing activity against 1-3 heterologous MV strains. A preponderance of neutralizing titers against heterologous MV strains by 1-log2 in some sera (6.5-32.5 depending on age) was most likely to suggest that the vaccinees' had been in contact with these virus strains in the past. In our investigation, a combination of positive IgG titers and neutralizing titers against the vaccine strain 2-log2 or higher provided the protection of the vaccinated children and teenagers against the symptomatic infection. There was a pronounced buster effect of the second immunization and a drop in the neutralizing activity of the sera from the vaccinated children and adolescents over time after the first and second immunization.

[Characteristics of humoral immunity to rubella virus in residents of Moscow in 1998-1999]. / Pokazateli gumoral'nogo immuniteta k virusu krasnukhi u zhitelei Moskvy v 1998-1999gg.

A total of 400 blood samples were collected from residents of Moscow in 1998-1999, 369 from adults aged mainly 19-31 years and from children aged 5-12 years. The mean incidence of antirubella antibody was 76.5%; the value varied in different age groups. The highest percentage of antibody detection was observed in men aged 19-25 years (91.8%) and the lowest in pregnant women aged 18-38 years (66-67%). Estimation of antihemagglutinin titers in international Units showed that antibody titer 1:40 and higher protected from infection; such titers were detected in 73.6% examinees. In pregnant women the level of immunological defense was very low (only 56%), which necessitates urgent vaccination of adolescent girls and young women. The results of EIA were compatible with the results of indirect hemagglutination test (IHAT), but EIA was much more sensitive in cases with low titers of IHAT.

[Russian recombinant vaccine against hepatitis B (results of controlled trials)]. / Otechestvennaia rekombinantnaia vaktsina protiv gepatita B (rezul'taty kontroliruemykh ispytanii).

Two yeast recombinant vaccines were compared: the Russian hepatitis B Combiotech vaccine in a dose of 20 mg and the Recombivax-HB vaccine in a dose of 10 mg manufactured in the USA (Merk, Sharp and Dohme Ltd.). Both agents were found safe and low reactogenic. After a course of 3 vaccinations with the Russian vaccine 92.5 to 97.5% subjects vaccinated after the 0-1-2 months and 0-1-6 months protocols had high levels of specific antibodies. In the group vaccinated with the American vaccine, antibodies to HBsAg were found in the sera of 95 to 100% subjects. These results demonstrate high immunological activity of both Russian and American vaccines.

[Enzyme indicator of the immunobiologic activity of antigens]. / Fermentnyi pokazatel' immunobiologicheskoi aktivnosti antigenov.

The test-system has been elaborated for the determination of the ability of different antigens in the cultures of the mouse peritoneal macrophages. Measuring an intracellular acid phosphatase activity, reflecting the extent of the cell activation, was taken as a principle of this test-system. With the help of this test-system ability of a few antigens (lipopolysaccharide, polysaccharide, glycoprotein, protein and low molecular weight substances--peptide and glycopeptide) have been studied. It is testified that above mentioned antigens possessed different effects on the acid phosphatase activity.

[The chromatography of yeast recombinant HBsAg in porous silicas]. / Khromatografiia drozhzhevogo rekombinantnogo HBsAg na poristykh kremnezemakh.

The results of the study of adsorption processes of recombinant HBsAg on various porous silicas are presented. The maximal yield of the recombinant antigen by absorption chromatography was observed on unmodified macroporous glass MPS-2000. A mean yield in this case was 85%. Silica C-80 modified with polyvinylpyrrolidone proved to be most acceptable for gel-penetrating chromatography. The adsorption and gel-penetrating chromatography on porous silicas proved to be an effective method for purification of recombinant HBsAg.

[Elastase and acid phosphatase activity in bronchoalveolar lavage fluid during chronic bronchitis]. / Aktivnost' élastazy i kisloi fosfatazy v bronkhoal'veoliarnom smyve pri khronicheskom bronkhite.

The dynamics of acid phosphatase and granulocytic elastase activities in bronchoalveolar washings of 40 patients with chronic catarrhal bronchitis was studied. In case of effective cure, the activities of these enzymes were increased shortly on the 10-13 days from beginning of the cure. In order to study the nature of this enzymes, the additional researches were conducted. These researches demonstrated that the macrophages were the source of the acid phosphatase, and the neutrophils--elastase. The transitory increase of the activity of both enzymes is favourable prognostical sign of disease's course.

[A stat-method of determining respiratory syncytial virus]. / Ekspress-metod opredeleniia respiratorno-sintsitial'nogo virusa.

For rapid detection of RS virus we have modified agglutination test with staphylococcus coated with RSV-antibody which allows the virus titer to be determined within 3-5 min. The results of RS virus titration in the yield compared with those obtained by CFT and CPE tests showed our modified test to be twice as specific and sensitive (60-80 ng/ml). This modification of the coagglutination test with sensitized staphylococcus and the method of running the test on a row of slides may be used in virological and serological laboratories as well as in infectious hospitals and outpatient wards.

[Isolation of viruses from water using porous silica]. / Vydelenie virusov iz vody s ispol'zovaniem poristykh kremnezemov.

The possibility of isolation of different viruses, hepatitis A virus among them, from water by means of adsorption chromatography on porous silica was demonstrated. Adsorption of viruses on the surface of silica depended on the kind of sorbent, water pH, and the rate of water flow through the column. The maximum elution of the adsorbed virus was observed using buffer solution with pH at least 11.0 and containing 1-2M NaCl. These conditions for adsorption-elution were found to be universal for different enterovirus types. In the field, the developed method was used for testing water specimens from various water bodies (sewage, river water, tap water). No viruses were found in 11 native specimens. Eluates of 4 out of 11 specimens were found to contain enteroviruses, hepatitis A virus among them.

[Immunological status of people inoculated with various types of inactivated vaccine against tick-borne encephalitis]. / Immunologicheskii status liudei, privitykh razlichnymi tipami inaktivirovannoi vaktsiny protiv kleshchevogo éntsefalita.

As compared with cellular immunity tests, serological tests more frequently detect virus-specific immune response in humans immunized with concentrated purified vaccine (CV) against tick-borne encephalitis. Combined determination of cell reactions and antibody production after a single injection of the vaccine increases the rate of detection of specifically responding persons. The duration of the inactivated vaccine-induced immunity is determined not only by the level of serum antibodies but also by the state of immunological memory detectable by the capacity of the host to produce antibodies after another injection of the specific antigen. At 14 and 30 days after a single immunization with CV IgG antibody is found in the blood of the vaccinees. In the course of 3 vaccinations, the concentration of serum immunoglobulins, chemotaxic activity of leukocytes, and blood lymphocyte reactivity to mitogens do not change. The role of some immunological values as criteria for the evaluation of the effectiveness of vaccine purification from impurities is discussed.

[Immunological indices of human vaccination against tick-borne encephalitis using inactivated preparations with a varying concentration of the viral antigen]. / Immunologicheskie pokazateli vaktsinatsii liudei protiv kleshchevogo éntsefalita inaktivirovannymi preparatami s raznoi kontsentratsiei virusnogo antigena.

The results obtained in the determination of the immunological activity of 3 types of formalin-killed tissue-culture tick-borne encephalitis vaccine, commercial non-concentrated and concentrated by 2 different methods, in humans are presented. Judging by the data of 3 serological tests and the blast transformation test, the concentration of killed tick-borne encephalitis virus enhanced immune response to vaccination. The intensity of this response correlated with the mouse protection capacity of the vaccine batches under test. An increase in the content of viral particles in the preparation is considered to be a promising approach to enhancing the effectiveness of vaccination against tick-borne encephalitis.

[Chromatography of formalin-inactivated tick-borne encephalitis viruses on macroporous glass]. / Khromatografiia inaktivirovannogo formalinom virusa kleshchevogo éntsefalita na makroporistykh steklakh.

The results of further development of methods for chromatographic concentration and purification of tick-borne encephalitis virus on columns with porous glass using formalin-inactivated virus suspensions are presented. The method of adsorption chromatography on porous glass under optimal conditions concentrates inactivated TBE virus 20-40-fold with simultaneous purification from protein by 93%-95%. Inactivated virus was shown to keep on glass better than infectious virus. Gel filtration chromatography removes 99.2%-99.8% of protein impurities from inactivated TBE virus preparations giving a complete or nearly complete "yield" of virus particles. The sequential use of adsorption chromatography and gel filtration produced concentrated, most highly purified TBE virus preparations containing no more than 2 microgram/ml protein. Chromatographic virus preparations were immunologically active in experiments in laboratory animals.