The immunomodulatory properties of immunobiological drugs Glutoxim and Phosprenyl we well as vesicular stomatitis virus and inactivated tick-borne encephalitis vaccine virus were studied using human diploid fibroblast cell line from the collection of M. P. Chumakov Federal Research Center for Research and Development of Immunobiological Products. All tested preparations exhibited immunomodulatory activity in human diploid fibroblast cell line. Glutoxim in doses of 0.1 and 0.25 μg/ml stimulated production of IL-6 and IL-10 during 24-48 h of culturing, but did not stimulate production of IL-1β. Phosprenyl, on the contrary, increased production of IL-1β and the levels of IL-6 and IL-10. Vesicular stomatitis virus stimulated the production of IL-1β, IL-6, and IL-10, while inactivated tick-borne encephalitis vaccine virus stimulated the production of cytokines IL-8 and IL-18. Immunomodulatory activity of inactivated tick-borne encephalitis vaccine virus was first demonstrated in the in vitro system.

We studied the sensitivity of domestic proprietary human and animal cell lines from the collection of M. P. Chumakov Federal Scientific Center for Research and Development of Immuneand-Biological Products to infection with different enterovirus 71 strains. A cell system based on domestic proprietary permanent cell line 4647 was for the first time used for reproduction of four enterovirus 71 strains (BrCr, 42266, 42934, and 43374). It was shown that strain 4647 is the optimal cell substrate for enterovirus 71 reproduction. The titers of enterovirus 71 for all four strains considerably (by 2 lgTCID 50 /ml and more) increased during sequential passages in permanent cell line 4647. The prospects of using permanent cell line 4647 for creation of diagnostic and preventive preparations against 71 was demonstrated.

Рассмотрена роль первичных фибробластов и линий диплоидных клеток фибробластов человека в формировании противоинфекционного иммунитета. Обсуждается адекватность клеточной модели на основе фибробластов для изучения молекулярных механизмов иммунопатогенеза вирусных инфекций. Рассмотрена возможность использования фибробластов для изучения особенностей действия иммунотропных препаратов. Обсуждается перспективность клеточной модели на основе аттестованных линий диплоидных клеток фибробластов для изучения механизмов действия новых и известных иммуномодуляторов.

Addition of adjuvants is a generally recognized method of enhancing the immunogenicity of vaccines. There is a large variety of adjuvants, therefore the choice of an adjuvant is based on the comparison of adjuvants efficacy in animal models, as well as assessment of their safety and tolerability. There are many different groups of substances that may have adjuvant properties, but relatively little attention is paid to carbohydrate-based adjuvants, despite the fact that they are compatible with live vector vaccines, safe, well tolerated, and their production is not laborious. Thanks to these advantages the use of these particular adjuvants in combination with any type of vaccine, including vector vaccines and DNA vaccines, may be most appropriate and promising. This review examines various types of carbohydrate adjuvants and the mechanisms that determine their efficacy.

The purpose of the study is the investigation of etiological, clinical and epidemiological features of HFRS, caused by Sochi virus in the Krasnodar area, as well as a comparative analysis of the data with those of HFRS, caused by Kurkino virus in the Central Russian regions and Puumala virus in the areas of Volga region. Materials and methods For the identification of HFRS patients, sera from more than 800 acute febrile patients residing in Krasnodar area were examined for hantavirus antibody by IFA with Puumala, Hantaan, Seoul, Sochi, Kurkino and Dobrava viruses. For primary screening there was used the indirect immunofluorescence (ELISA) method with polyvalent cultural antigen as for serotyping of positive sera according to affiliation of antibodies to various Hantaviruses species, there were used ELISA method with monovalent cultural antigens and the neutralization reaction with Puumala, Hantaan, Seoul, Sochi, Kurkino and Dobrava viruses with the use of the method of Inhibition of Focus-Forming Units assay. Clinical and epidemiological studies have been performed on the base of history cases and records of the epidemiological examination. Results of the study. During 2000 - 2013 there were identified 64 patients suffered from HFRS caused by the Sochi virus. The patients resided in 36 settlements of 10 administrative districts of the Krasnodar area. A comparative analysis of clinical, laboratory and epidemiological data of patients with HFRS-Sochi, HFRS-Kurkino and HFRS-Puumala viruses allowed to reveal differences between the clinical (frequency of registration and severity of several symptoms, severity of the course and mortality rate) and epidemiological (prevalence in rural and urban residents, occupational pattern, the seasonality of the disease, conditions of contamination) manifestations. Conclusion There was established the etiological and epidemiological importance of Sochi virus. Sochi virus causes sporadic annual incidence of HFRS in the territory of Krasnodar area. Cases of HFRS caused by the Sochi virus are differ in more severe course of the disease and high lethality rate in comparison with the other two forms of HFRS caused by Puumala and Kurkino viruses in the territory of the European part of Russia.

The work is devoted to clarify the etiological aspects of the hemorrhagic fever with renal syndrome (HFRS), clinical and epidemiological features that are closely linked to the etiology, as well as to development of methods and products for specific diagnosis and vaccine prevention of hantavirus infection. That regard to hantaviruses a set of virological, immunological and molecular-genetics methods were developed significantly enhance the effectiveness of the specific diagnostics of HFRS. The features of humoral immunity in HFRS were identified and atypical clinical forms of HFRS was istablished; indicators of natural immunity to hantavirus in the population living in different regions of Russia and the former Soviet republics have been identified that allowed us to refine nosological area of HFRS; new, previously unknown natural foci of HFRS, including hantavirus RNA, in the central regions of Russia and in the subtropical zone of Krasnodar region have been identified and studied; 76 strains and 70 isolates of 9 rodent species and one species of birds, as well as from the blood of patients with HFRS and sectional materials from HFRS dead patients have been isolated and identified; new hantavirus species - Khabarovsk, Taimyr-Topografov, Adler, as well as two new genotypes Dobrava/Belgrad virus - Kurkino and Sochi have been identified; species composition of small mammals - the natural reservoir of hantaviruses was the refined; etiological role and epidemiological importance of different hantaviruse types in HFRS incidence structure was established; the existence of new etiologically distinct hantavirus infections with significant epidemiological differences has been proven; manufacturing techniques and methods of control of the culture inactivated vaccine against HFRS has been developed.

The work is devoted to clarify the etiological aspects of the hemorrhagic fever with renal syndrome (HFRS), clinical and epidemiological features that are closely linked to the etiology, as well as to development of methods and products for specific diagnosis and vaccine prevention of hantavirus infection. That regard to hantaviruses a set of virological, immunological and molecular-genetics methods were developed significantly enhance the effectiveness of the specific diagnostics of HFRS. The features of humoral immunity in HFRS were identified and atypical clinical forms of HFRS was istablished; indicators of natural immunity to hantavirus in the population living in different regions of Russia and the former Soviet republics have been identified that allowed us to refine nosological area of HFRS; new, previously unknown natural foci of HFRS, including hantavirus RNA, in the central regions of Russia and in the subtropical zone of Krasnodar region have been identified and studied; 76 strains and 70 isolates of 9 rodent species and one species of birds, as well as from the blood of patients with HFRS and sectional materials from HFRS dead patients have been isolated and identified; new hantavirus species - Khabarovsk, Taimyr-Topografov, Adler, as well as two new genotypes Dobrava/Belgrad virus - Kurkino and Sochi have been identified; species composition of small mammals - the natural reservoir of hantaviruses was the refined; etiological role and epidemiological importance of different hantaviruse types in HFRS incidence structure was established; the existence of new etiologically distinct hantavirus infections with significant epidemiological differences has been proven; manufacturing techniques and methods of control of the culture inactivated vaccine against HFRS has been developed.

There are presented results of the adaptation ofpathogens of hemorrhagic fever with renal syndrome (HFRS) virus Puumala and Dobrava strains, to the reproduction in cell cultures of the simian origin - Green monkey kidney (Vero, 4647), Green monkey spleen (455), calf kidney (PT-1) and sheep kidney (4184). Hantaviruses were shown to be able to adapt to the replication in the new cellular system differing in species origin from the culture of Vero E6 origin in which the strains were isolated. This fact demonstrates the heterogeneity of hantavirus population, with the existence of «quasispecies» which can recognize and bind to ligands (b3 integrins) of the other species specificity. The highest yield of the virus, which is comparable with the culture of Vero E6, culture Vero provides. In Vero cells after 3-4 passages Puumala and Dobrava-Belgrade viruses replicate also actively as in a permissive culture Vero E6, that proposes the use of Vero culture cell allowed for the production of immunobiological drugs administered to people as a substrate in the delivery of vaccines against HFRS.

The monoclonal antibodies to Puumala, Dobrava, Hantaan, and Seoul hantaviruses were obtained using mice. The viruses were known to cause HFRS, and two variants of ELISA were designed. First, Hanta-PUU variant, was constructed using monoclonal antibodies to Puumala virus envelope glycoprotein (G(N):G(C)) for detecting only Puumala virus antigen. The second, Hanta-N variant, was constructed using monoclonal antibodies to Dobrava and Puumala nucleocapsid proteins for detecting four above mentioned hantaviruses. Both Hanta-PUU and Hanta-N assays were reliable in detecting specific hantavirus antigens and the immunogenecity of hantavirus vaccines.

The monoclonal antibodies to Puumala, Dobrava, Hantaan, and Seoul hantaviruses were obtained using mice. The viruses were known to cause HFRS, and two variants of ELISA were designed. First, Hanta-PUU variant, was constructed using monoclonal antibodies to Puumala virus envelope glycoprotein (G N:G C) for detecting only Puumala virus antigen. The second, Hanta-N variant, was constructed using monoclonal antibodies to Dobrava and Puumala nucleocapsid proteins for detecting four above mentioned hantaviruses. Both Hanta-PUU and Hanta-N assays were reliable in detecting specific hantavirus antigens and the immunogenecity of hantavirus vaccines.

The findings suggest that there are natural foci of hantavirus infection in the Tambov Region. There is evidence that Dobrava/Belgrade hantavirus (DOB-Aa) was a leading etiological agent in the outbreak of the disease in the winter of 2006-2007. Epidemiological analysis showed that the outbreak of hemorrhagic fever with renal syndrome (HFRS) afflicted the region during November to April, by reaching its peak in January (52.2%). Among the patients with HFRS, rural dwellers were 91%. People were infected with the virus mainly by taking care of domestic animals (97.2%). The reservoir of the virus and the source of its human infection in the outbreak was a field mouse, its western subspecies Apodemus agrarius agrarius, which was absolutely dominated among all the virus carriers.