[The capacities of current test systems to verify early HIV infection].

The purpose of the present investigation was to comparatively evaluate the performance characteristics of the test systems designed to verify the positive results of screening survey for HIV infection, such as the solid-phase immunoassay DS-EIA-HIV-AB/AG-SPECTR (Diagnosticheskiye Sistemy (Diagnostic Systems) Research-and-Production Association, Nizhni Novgorod) and tests based on immune blotting (IB). The investigation examined 15 seroconversion panels produced by ZeptoMetrix (USA) and BBI (USA). The use of the DS-EIA-HIV-AB/AG-SPECTR test system determined 88 of the 167 seroconversion panels as HIV positive. The IB-based tests revealed only 45 of the 167 samples as positive. Consequently, the application of the DS-EIA-HIV-AB/AG-SPECTR test system is more effective than the IB-based tests in early HIV infection.

[Vitaherpavac is the first Russian herpes simplex virus vaccine obtained on the Vero B continuous cell line].

Vitaherpavac, a dry inactivated herpes simplex virus (HSV) culture vaccine, has been obtained, by using the Vero B continuous cell line as a substrate for accumulation of herpes simplex virus types 1 (US strain) and 2 (VN strain). Vitaherpavac and the similar vaccine Herpovax made by the Research Institute of Vaccines and Sera, Saint Petersburg (for which preparation a primary trypsinized chick embryo cell culture used as a substrate for accumulation of HSV types 1 and 2), underwent comparative clinical trials. The tolerability and therapeutic effectiveness of the vaccine were tested in patients diagnosed as having chronic frequently recurring herpes. The trials have yielded positive results that suggest that it is expedient to introduce of the new vaccine Vitaherpavac into practice to treat chronic recurrent herpetic infection of various localizations. Vitaherpavac has been registered in the Russian Federation and permitted for medical application.

[Vaccines, immunoglobulins, and test systems for the prevention and diagnosis of tick-borne encephalitis].

The quality of drugs used in the Russian Federation to prevent and diagnose tick-borne encephalitis (TBE) was analyzed. The TBE vaccines made in Russian were shown to be as effective as those manufactured in foreign countries and to have the similar production techniques and quality indices therefore they may be interchangeable in the population's vaccination. The specific activity of has recently increased in terms of the content of TBE virus antibodies. In Russia, a wide range of enzyme immunoassay systems have been designed and manufactured for the detection of human serum (plasma) IgM and IgG antibodies to TVE virus and for that of TBE viral antigen in the biological objects, primarily in the ticks.

[Comparative study of hemostasis and cytokine production in experimental Dengue virus infection].

Some parameters of hemostasis and cytokine production during the fatal and nonfatal course of experimental human versus murine infection caused by Dengue virus (DV) were studied. Its lethal dose administration induced a rapid development of anemia, thrombocytopenia and a change in packed cell volume. These changes were not so profound in the nonfatal group of infected mice. There was an excess production of both proinflammatory (TNF-alpha, IL-1beta) and anti-inflammatory (IL-6, IL-10) cytokines in the fatal group. In mice of this group, the early rise of IFN-gamma and IL-12 corresponded to the parameters observed in patients with severe forms of DV infection. The findings provide evidence that the proposed model may be used for the experimental study of the immunopathogenesis of the disease.

[ELISA and PCR test systems used to detect Crimean-Congo hemorrhagic fever virus].

Studying the sensitivity and specificity of enzyme-linked immunosorbent assay (ELISA) for the indication of Crimean-Congo hemorrhagic fever (CCHF) virus antigens and those of reverse transcription and polymerase chain reaction (RT-PCR) for the detection of CCHF virus RNA, and those of a intercerebral infection method in newborn albino mice systems for the determination of viral infectious activity established that the sensitivity of ELISA was 1-2 orders of magnitude less than that of RP-PCR. The latter proved to be better in studying the sera sampled from patients with CCHF. The results of studying the samples of H. marginatum ticks, the CCHF virus vectors by ELISA and RT-PCR were similar.

[Immune response in vaccinated and non-vaccinated mice infected with tick-borne encephalitis virus].

The paper deals with an investigation of an immune response in BALB/c mice immunized with tick-borne encephalitis (TBE) vaccine and infected with TBE virus and in non-immunized mice. The parameters of specific humoral (IgG and IgM) and cellular (gamma-interferon (IFN) and cell proliferation) immunities and the activity of cytokines (necrosis tumor factor-alpha, interleukin (IL)-1beta, IL-2, IL-6, IL-10, and IL-12) were studied. There were significant differences in the specific and nonspecific immune response of immunized and non-immunized animals. Noteworthy is the difference in the time course of changes in the levels of IL-6, IL-2, IL-12, and gamma-IFN in the immunized and non-immunized animals.

[Adaptation of different modifications of enzyme immunoassay and immune blot assay to assess the candidate vaccine HIVREPOL against HIV/AIDS].

A preclinical trial of the vaccine HIVREPOL provided a complex of methods for assessing the identity and specific activity of vaccines against HIVIAIDS. The identity of "HIVREPOL" has been assessed by indirect enzyme immunoassay (EIA): the vaccine specifically binds the antibodies of the sera from HIV-infected individuals. Immune blot assay was the most informative method for assessing the identity of the candidate vaccine. The sera from HIVREPOL-vaccinated mice recognized the proteins gp41, p24, p55 of cultured HIV1 on "New-Lay-Blot1" strips. The bands corresponding to p24 were revealed in the line blots "Blot-HIV-1/2+O" and "INNO-LIA-HIV-Confirmation". The specific activity of the HIVREPOL vaccine was confirmed from the reactivity of sera of the mice vaccinated with recombinant proteins of the immunosorbents available in EIA test systems for the detection of HIV antibodies. Competitive EIA established the antigen-binding activity of sera from HIVREPOL-vaccinated mice against the native reference HIV-1 antigen.

[Development of sera reference panels for quality control of ELISA diagnostic kits in Russia]. / Razrabotka standartnykh panelei syvorotok dlia kontrolia kachestva immunofermenthykh test-sistem v Rossii.

In the past 5 years, the investigators of the "VECTOR" SRB VB and the L.A. Tarasevich State Institute of Standardization and Control of Medical Biological Preparations have jointly designed sera reference panels containing anti-HIV-1 IgG, anti-HCV IgG, and anti-HAV IgM which have been approved as national standard panels. The panels are intended for use in controlling the specificity and stability of the most widely used ELISA diagnostic kits and immunoblot test systems during production, control, and application stages. Some problems of development and production of these panels, including the representation of different sera in the panels and the selection of specific IgG concentrations in the different sera in the panel are described. The authors also attract attention to the stabilization of the specific characteristics of panel sera during storage and transportation.

[Experimental infection caused by attenuated strains of viruses of the tick-borne encephalitis complex in the bodies of immunosuppressed animals]. / Izuchenie éksperimental'noi infektsii, vyzvannoi attenuirovannymi shtammami virusov kompleksa kleshchevogo éntsefalita v organizme immunodepressirovannykh zhivotnykh.

Exacerbation of virus infection caused by 4 attenuated strains of viruses of the tick-borne encephalitis complex in BALB/c mice treated with an immunosuppressant was demonstrated. The influence of the treatment was manifested both in an increased extraneural activity of the virus, higher mortality of the experimental animals, and in high virus titers in the brain, liver, and spleen of the immunosuppressed animals, and a decrease or complete absence of specific antibody in their sera.

[Activity of cytokines in immunization with a vaccine against experimental tick-borne encephalitis]. / Aktivnost' tsitokinov pri immunizatsii vaktsinoi protiv kleshchevogo éntsefalita v éksperimente.

An experimental double immunization of BALB/c mice with a vaccine against tick-borne encephalitis was accompanied by the production of IL-1b, IL-2, IL-4, IL-6, IL-10, IL-12, TNFa and gamma-IFN in the blood serum of animals. After the first immunization of animals with the vaccine against tick-borne encephalitis, the activity of cytokine was indicative of an activation of macrophages as well as of an activation and differentiation of T- and B-lymphocytes. After the second immunization and unlike the first one, there were changes in the production only of IL-10, IL-12 and gamma-IFN, which can be indicative of a regulation of balance between Th1 and Th2.

[The evaluation of a commercial EIA test system for determining the specific activity of antiherpetic preparations]. / Otsenka kommercheskoi test-sistemy IFA dlia opredeleniia spetsificheskoi aktivnosti protivogerpeticheskikh preparatov.

The data on the use of a commercial EIA test system for detection of antibodies in control of preparations against herpes simplex and cytomegaloviruses are presented. The enzyme immunoassay test system for antibody determinations to herpes simplex virus produced by the Odessa bacterial preparations enterprise was shown to be suitable for determination of the specific potency (antigenicity) of herpes simplex vaccine. The advantages of this method over the currently used neutralization test were established. Titration of commercial immunoglobulins detects lots with high litres of antibody to herpes simplex virus. For the same purpose, lots of commercial immunoglobulins were tested for antibodies to cytomegalovirus using a West Germany test-system (Behring). It is concluded that enzyme immunoassay test systems for antibody determinations may be used for screening of lots of immunoglobulins of special effects (against herpes simplex and cytomegalovirus infections) both at the stage of serum and final preparation screening.

[Survival of tick-borne encephalitis complex viruses in a brain cell culture from suckling mice]. / Perezhivanie virusov kompleksa kleshchevogo éntsefalita v kul'ture kletok mozga myshei-sosunkov.

The evidence of long-term survival of the tick-borne encephalitis complex viruses in primarily trypsinized brain cells of suckling mice inoculated in vivo is presented. The viability of the brain cell cultures from suckling mice inoculated with an attenuated strain was shown to be higher than that of a similar culture of the brain cells from suckling mice infected with a virulent strain. The infectious virus could be recovered for over 200 days of the existence of these cultures. The experimental results appear to be another confirmation of the capacity of viruses of the tick-borne encephalitis complex, particularly of the attenuated strains, for persistence.

[Use of inbred mice for assessing the neurovirulence of strains of the tick-borne encephalitis virus complex and the immunogenicity of a concentrated tick-borne encephalitis vaccine]. / Ispol'zovanie inbrednykh myshei dlia otsenki neirovirulentnosti shtammov virusov kompleksa kleshchevogo éntsefalita i immunogennosti kontsentrirovannoi vaktsiny kleshchevogo éntsefalita.

The sensitivity of different lines of inbred mice to strains of the tick-borne encephalitis complex viruses differing in their biological properties and the degree of attenuation was compared. The mice differing in their genetic characteristics and susceptibility to extraneurally inoculated virus were shown to have different immunological responsiveness to inoculation of an inactivated vaccine antigen.

[Immunoenzyme method in the diagnosis of tick-borne encephalitis]. / Immunofermentnyi metod v diagnostike kleshchevogo éntsefalita.

Data are presented on the use of direct and indirect enzyme-immunoassay (EIA) in the study of tick-borne encephalitis (TBE) virus. EIA has been shown to be considerably more effective than the conventional CFT and HI tests exceeding them in sensitivity by 50-150 times both in studies of the antigenic activity of various preparations and in examinations of human sera. The possibility of testing various antigenic substrates makes EIA a universal method suitable for numerous studies. The high sensitivity of the method is useful for antigenic differentiation of viruses and strains within the TBE complex. The EIA may be used satisfactorily for early diagnosis of TBE and study of seroconversion in vaccines.

[Quantitative indices of the immune status of persons inoculated with different vaccines against tick-borne encephalitis]. / Izuchenie nekotorykh kolichestvennykh kokazatelei immunnogo statusa liudei, privitykh razlichnymi vaktsinami protiv kleshchevogo éntsefalita.

Inactivated concentrated and unconcentrated vaccines against tick-borne encephalitis given to 80 normal subjects produced no suppressing effect on the quantitative values of the immune status. Immunogenesis was characterized by stimulation of both T- and B-links of the immune system of the vaccines. With the unconcentrated vaccine a greater response was demonstrated after 3 injections; the concentrated vaccine induced an intensive immune response after 2 injections.

[Collection of pathogenic viruses: state of the problem and prospects]. / Kollektsionirovanie patogennykh virusov: sostoianie i perspektivy problemy.

The organizational and methodological separateness in the collection of pathogenic viruses impedes the development of universal approaches to practical collection. The authors propose a universal research program for the creation of specialized National Museums of Pathogens as reference and methodological centers within the framework of the Russian State Collection of Microorganisms. Further improvement of methods for virus collection is desirable.

[Design of reference panels of serum for testing the quality of HIV antibody tests in practical laboratories]. / Konstruirovanie otsenochnykh panelei syvorotok dlia testirovaniia kachestva provedeniia analizov na antitela k VICh v prakticheskikh laboratoriiakh.

The authors define the scientific basis for development of panels of reference sera intended for effective control of the quality of enzyme immunoassay of HIV antibody screening. Special attention was paid to developing the technology of preparing standards highly stable serum samples with a preset concentration of anti-HIV antibodies. The resultant panel of reference sera was tried in screening for anti-HIV antibodies by enzyme immunoassay at seven diagnostic laboratories. Mathematical analysis of the results permits the detection of the minimal errors in studies with the use of serum panel of practical laboratories.

[Preparation of rabbit antiserum to Ebola virus]. / Poluchenie krolich'ikh antisyvorotok v virusu ébola.

Humoral immunity of rabbits insusceptible to Ebola virus infection were studied after challenge with infectious Ebola virus and inactivated antigen of this virus. Administration of Ebola virus antigen induced the production of specific antibodies in proportion with the antigen dose injected.

[Comparative study in an experiment on the efficacy of 2 regimens of vaccinal immunization against human acute encephalomyelitis]. / Sravnitel'noe izuchenie v éksperimente éffektivnosti dvukh skhem immunizatsii vaktsinoi protiv ostrogo éntsefalomielita cheloveka.

Experimental data on the effectiveness of two schedules of vaccination against human acute encephalomyelitis, one conventional, using 12 injections, and another shortened, using 8 injections, are presented. The experiments in inbred mice evaluating the general resistance, virus-neutralizing antibody titres, resistance in adoptive transfer tests, and values of lymphocyte blastogenesis test showed no statistically significant differences between the shortened vaccination schedule and the conventional one. Experimentally, the shortened vaccination schedule is as effective as the conventional one.

[Development and application of an immunoenzyme test system for diagnosing Ebola fever]. / Razrabotka i primenenie immunofermentnykh test-sistem dlia diagnostiki likhoradki ébola.

Enzyme immunoassay (EIA) test systems for the detection of antigens of and antibodies to Ebola virus were developed and tried. The test system for the detection of Ebola virus antigens based on direct solid-phase EIA detects viral antigens in culture fluid of infected Vero cells, in the blood sera, and in homogenates of infected tissues. Use of this test system allows detection of at least 10 ng of viral proteins or 5.0 x 10(3) to 1.0 x 10(4) PFU/ml in infectious material. The test system is prepared on the basis of protein A - horseradish peroxidase conjugate. It is universal for the testing of animal and human sera and is characterized by high resolution and reproducibility of results. It allows detection of antibodies to Ebola virus starting from days 8-9 of infection. A higher sensitivity of direct solid-phase EIA in comparison with complement fixation or indirect immunofluorescence tests is demonstrated.