Immunobiological Preparations Research Articles

Use of MONTANIDE ISA 78 VG as adjuvant for the manufacture of antibacterial vaccines for chickens

Relevance. The use of antimicrobial agents and vaccine prophylaxis are the main ways to prevent and control most diseases of bacterial etiology. At the same time, uncontrolled use of antimicrobial agents without determining the sensitivity of microorganisms, as a rule, does not give the opportunity to obtain a positive therapeutic result. At the same time, a competently designed scheme of inactivated vaccine administration, taking into account the epizootic situation, is an effective and safe tool for controlling bacterial diseases.In this regard, the results of testing of vaccine samples against bacterial diseases of birds based on the modernized oil adjuvant Montanide ISA 78 VG presented in this article are interesting and modern.Methods. Four vaccine samples were manufactured for research based on the oil adjuvant Montanide ISA 78 VG. The first sample of the vaccine is against avian salmonellosis, the second — against avian pasteurellosis, the third — against avian respiratory mycoplasmosis, the fourth — against avian hemophilosis. All manufactured vaccine samples were investigated for sterility, stability and viscosity of the emulsion according to generally accepted methods. To determine the reactogenicity and antigenic activity of the vaccine samples, young egg-laying hens aged 25–40 days were used.Results. Analysis of the obtained results of physical and biological properties of tested inactivated vaccines against bacterial diseases of birds allows us to conclude that the vaccine against respiratory mycoplasmosis of birds made on the basis of adjuvant Montanide ISA 78 VG is a safe and effective immunobiological preparation.

Experimental immunotherapy of association infections of the gastrointestinal tract of newborn calves using yolk melange enriched with specific Y class immunoglobulins

Mass gastrointestinal diseases of newborn calves manifested by symptoms of diarrhea are caused by various etiological factors and most often occur in the form of mixed infections. Mixed infections have a more severe diarrhea than mono-infections. The economic consequences of such infections due to the high prevalence, frequent mortality and loss of productivity during further exploitation of the animal can be significant. The most common causative agents of gastrointestinal diseases of newborn calves of viral etiology is rotavirus and coronavirus infections. Mixed (viral-bacterial) infection can lead to 100% mortality of the diseased calves. Specific vaccination of springer cows against gastrointestinal pathogens in calves, which is performed to increase the number of pathogen-specific immunoglobulins in the colostrum at calf birth, and ingestion of colostrum by the calves during the first 4 hours of life are the most important strategies to prevent severe diarrhea. But as practice shows, this is not always enough. The purpose of this work was to study the effectiveness of immunoprophylaxis of associative infections of the gastrointestinal tract of newborn calves with the use of melange prepared from chicken eggs previously immunized with the vaccine "Rotagal" (Pharmagal-BIO s.r.o., Slovakia), used for the prevention of colibacillosis, rotavirus and coronavirus infections of cattle. As a result of the study, it was established that the addition of yolk melange rich in specific immunoglobulins of Y class to the colostrum in the first hours of life of the calves has a pronounced prophylactic effect, facilitates the course of the infectious process in case of the disease, saves costs for the treatment of sick animals, increases survival rate, contributes to the acceleration of live weight gain. The use of an immunobiological preparation based on transovarial immunoglobulins has a positive effect on the indicators of nonspecific resistance of newborn calves compared to the calves with early postnatal disease and antibiotic treatment.

Isolation of donor gamma globulin obtained from multiparous women and its effects upon expression of HLA-G and HLA-DR molecules on lymphocytes from mothers of children with septal congenital heart defects

A common pathogenetic mechanism of reproductive losses and congenital heart disease (CHD) is associated with immune inflammation in the “mother-embryo” system which affects differentiation and proliferation of cardiovascular progenitor cells. It is hypothesized that this link may be blocked by regulatory auto- and alloimmune antibodies to HLA-G and HLA-DR molecules. Moreover, these antibodies may be present at sufficient amounts in donor immunoglobulins, especially those obtained from the blood of multiparous women. Based on this suggestion, the aim of our study was to obtain enriched gamma globulin fraction from the blood of multiparous women and evaluate its functional effects towards HLA-DR and HLA-G molecules. Isolation of the gammaglobulin fraction (GGF) from the blood plasma of multiparous women was performed using affinity chromatography in several sessions. Purity grade of the resulting protein was analyzed by immunoelectrophoresis, electrophoretic separation of the protein fraction of blood serum and electrophoresis in 4.12% polyacrylamide gel with the addition of SDS (PAGE electrophoresis). PAAG electrophoresis showed that this GGF did not differ from commercial therapeutic intravenous immunoglobulin (IVIG). Assessment of the functional activity of GGF upon HLA-DR and HLA-G molecules was performed in the main group of women and their children with congenital heart disease (n = 38), and control group of women who gave birth to conditionally healthy children (n = 21). To determine the specificity of GGF with respect to HLA-G, HLA-DR molecules, as well as to compare its effect with autologous and allogeneic sera and IVIG, we developed an immunological testing protocol using flow cytometry. The protocol was arranged on the basis of the methodology of “cross-match” approach and Russian patent “Method for determining antibodies to HLA-G”. It was found that the blocking activity of female serum towards autologous (intrinsic) and allogeneic (embryo/fetus/child) HLA-G and HLA-DR molecules may determine the protective effect on development of congenital heart defects in the next generation. Donor human immunoglobulin showed a similar blocking effects to these molecules, possibly due to the presence of alloimmune antibodies to HLA classes I and II. The gammaglobulin fraction obtained from the donor blood of multiparous women has a more pronounced blocking effect on the HLA-G and HLA-DR expression. Hence, this immunobiological preparation can be considered a prototype of therapeutic and prophylactic agent blocking the genesis of congenital heart defects.

Electrochemical Immunosensors Based on Gold Nanoparticles for the Determination of Ovalbumin in Immunobiological Preparations

Electrochemical Immunosensors Based on Gold Nanoparticles for the Determination of Ovalbumin in Immunobiological Preparations

Differences in complete blood count parameters in patients with COVID-19 diagnosis based on vaccination status (literature review)

Relevance. Due to the large-scale vaccination campaigns against the SARS-CoV-2 coronavirus taking place worldwide, the question arises as to how vaccination may affect complete blood count (CBC) parameters in COVID-19 cases. Research objective: assess differences in complete blood count parameters in COVID-19 patients based on their vaccination status. Materials and methods. A review of publications was conducted describing and presenting the complete blood count parameters in patients with verified COVID-19, by stratifying them into vaccinated and unvaccinated individuals against the SARS-CoV-2 virus-caused disease. Subsequently, a meta-analysis of the results for each CBC parameter was carried out by describing identified differences among patients with mild and moderate COVID-19, lacking comorbidities affecting CBC indicators. Results. The analysis conducted allowed to determine that the complete blood count parameters do not exceed the reference range in most cases in both groups, excepting those of individuals with a severe disease course. The exception is the erythrocyte sedimentation rate (ESR) (42 [20.5–63.5] (∑n = 107, Me [Q1–Q3])) mm/h in unvaccinated individuals. Also, a slightly lower absolute lymphocyte count is observed in vaccinated individuals (0.95 [0.58–1.62] × 109/L (∑n = 441, Me [Q1–Q3])). There is a twofold greater range of leukocyte quartile values in vaccinated individuals compared to unvaccinated individuals: 7.07 [4.07–12.31] × 109/L (∑n = 555, Me [Q1–Q3]) vs 5.68 [4.02–8.34] × 109/L (∑n = 2202, Me [Q1–Q3]). A similar pattern is observed for monocytes. Conclusion. Conducting a meta-analysis of laboratory parameters in COVID-19 cases among vaccinated and unvaccinated individuals allows for determining the effectiveness of vaccination as well as identify the major trends in dynamic changes for essential blood parameters among unvaccinated and vaccinated individuals in general or with certain immunobiological preparations.

DIRECTIONS OF DEVELOPING NEW TECHNOLOGIES BASED ON CULTIVATION OF ANIMAL CELLS AND TISSUES

Animal cell cultures have found applications in various fields, from basic to advanced research. This includes studying the fundamentals of cell biology, mechanisms of the cell cycle, specialized cell functions, cell-cell and cell-matrix interactions, toxicity testing for the study of new drugs, gene therapy for replacing non-functional genes with functional cells, characterization of cancer cells, understanding the role of various chemical substances, viruses, and radiation in cancer cells, vaccine production, monoclonal antibodies, and pharmaceuticals. Additionally, the cultivation of viruses for use in vaccine production, such as for diseases like rabies, hepatitis B, and measles, is another important application of animal cell cultures. Aim. The purpose of this study was to analyze the literature data on the use of animal cell lines in genetic engineering, therapy, xenotransplantation, biopharmaceuticals, the food industry, and research. Methods. An analytical review of literature data was conducted using the information analysis of Medline (PubMed), Web of Science and Scopus databases, Google Scholar, the Cochrane Central Register of Controlled Trials (CENTRAL), and other sources up to the inclusive year 2023 using the keywords: “animal cell lines”, “immunobiological preparations”, “xenotransplantation”, “biopharmaceuticals”, “genetic engineering”. Results. An analysis of research related to the use of animal cells in the biopharmaceutical industry was carried out, and considerations regarding the prospects for their use in various research and production technologies were outlined. Conclusion. The technology of cultivating animal cells has become a fundamental tool in the development of research in the field of biotechnological sciences. The ability to culture animal cells in vitro has allowed the development of innovative methods, such as iPSC and organ-on-a-chip models, which have provided valuable information about disease mechanisms and potential therapeutic targets. Although there are some challenges with the use of animal cells related to variability in differentiation efficiency and concerns about safety and efficacy, further studies are needed to optimize protocols and overcome these limitations. Overall, animal cell culture technology remains an important component of modern biomedical research and has the potential to revolutionize the field of regenerative medicine.

Перспективы применения иммуно-ПЦР в диагностике SARS-CоV-2

One of the main biological characteristics of SARS-CoV-2 is its remarkable ability to adapt to new environmental conditions. Accelerated evolution of the COVID-19 pathogen complicates anti-epidemic measures, including the need to update the antigenic composition of vaccines and maintain current vaccination measures, complicates the prognosis of the infection course, and reduces the efficacy of drugs based on monoclonal antibodies and specific antiviral inhibitors. Under these conditions, the search for new approaches to monitoring clinically and epidemiologically significant variants of the virus that escape the factors of humoral immunity, as well as to the accelerated development of relevant immunobiological preparations, becomes important. This in turn requires the use of approaches to assess the serological properties of new variants, such as neutralization reactions using live infectious virus or immunochemical methods with limited antigen detection. Immuno-PCR (iPCR) is a method which is lacked majority of virus neutralization reaction and immunochemical methods (ELISA and LFT) disadvantages. The iPCR method has many modifications aimed at increasing specificity while reducing background amplification, simplifying staging, and multiplexing. Taking into account the high sensitivity of iPCR, it is possible not only to measure the exact amount of antigen, but also to use the diagnostics in the minimum quantities available, for example, in clinical samples. This fact makes iPCR method high-promising in personalized medicine field. Key words: SARS-CoV-2, coronavirus evolution, immuno-PCR, humoral immunity

Some reflections on immunosuppressants and their delivery to the lungs by aerosol in the treatment of COVID-19

Disorders of immune homeostasis represent the key pathogenetic link of COVID-19, which often manifests as a hyperimmune response to the pathogen, leading to severe uncontrolled inflammation in lungs, followed by complications and death. Accordingly, a certain therapeutic potential is provided by different pharmacological drugs with distinct mechanisms of action, This class of drugs should, however, act in common direction by suppressing the immune response, thus being often classified as immunosuppressants (IS). Of them, the most promising are immunobiological preparations, which include monoclonal antibodies, as well as purinergic regulatory agents. There are several attempts to use the “classical” IS by a certain way, e.g., cytostatics and calcineurin inhibitors which found clinical application in transplantology and oncology. However, their usage for treatment of uncontrolled inflammation of respiratory tract was abandoned by the end of XX century. Meanwhile, the aerosol route of drug administration optimizes treatment, both in terms of their effectiveness, and the reduction of side effects thus promoting usage of IS for treatment of uncontrolled airway inflammation. Previously, the analysis of therapeutic opportunities for some IS delivered as aerosols to the lungs in COVID-19 therapy was not carried out, thus bein the purpose of our work. Methodological analysis was carried out using various databases of biomedical scientific information, including Index Medicus, PubMed, Embase, Cohrane Clinical Trials gov registry and patent databases.The efficiency of the impact of various IS subgroups in COVID-19, including their administration by inhalations into the respiratory ways, was assessed. The role of regulatory T cells considered the central regulator of immune response, in pathogenesis of COVID-19 was considered, and their therapeutic potential was characterized, dependent on phase and severity of the disease as well as drug dose dependence. Methods and approaches to the use of IP, advantages and disadvantages are discussed. The expediency and future prospects of their application are considered.One may conclude that the effectiveness of cytostatics and calcineurin inhibitors in the treatment of airway inflammation in COVID-19 remains unconfirmed and seems to be unpromising. Meanwhile, biological preparations, including monoclonal antibodies and purinergic regulatory agents, offer great promise in this respect.

DEVELOPMENT OF THE TECHNOLOGY OF DEEP CULTIVATION OF A TRANSFORMED B. SUBTILIS STRAIN

Deep cultivation of recombinant cultures in bioreactors makes it possible to accumulate a large number of specific antigens used in the production of immunobiological preparations and diagnostic test systems in a short period of time, taking into account stimulating and limiting growth factors. The main purpose of the research was to determine the influence of the factors of seed concentration, pH level and the level of dissolved oxygen on deep cultivation, with the control of accumulation kinetics by turbidimetry. During the research, it was found that the most optimal inoculation concentration is 1 million cells/cm3 with a maximum final concentration of B. subtilis 4x106 cells/cm3, the most optimal pH level is a dynamic mode from 8.0 to 6.0 with a maximum final concentration of 5x106 cells/cm3, the most optimal level of dissolved oxygen is stationary mode with a minimum oxygen level of 25%, with a maximum final concentration of 4x106 cells/cm3.Bacteriological, technological and biochemical methods were used in the work to develop the modes of deep cultivation of the transformed B. subtilis strain with control of pH parameters and the level of dissolved oxygen.The scientific significance of the research is based on the development of deep cultivation, with an assessment of external growth factors and their influence on the kinetics of accumulation of the transformed B. subtilis strain, which will provide effective conditions for the growth and production of microorganisms. The practical significance of the research is based on the possibility of optimizing the biotechnological stages of the cultivation of the transformed B. subtilis strain, which will positively affect the efficiency of submerged cultivation as the main method for the accumulation of specific antigens.

Prevention methods for clostridial infection of cattle on the territory of the Russian Federation

This article presents a review of the literature data of foreign and domestic scientists, as well as an analysis of the results of our own research on the problem of clostridial infections in cattle. This paper provides insight into current etiological structure of clostridiosis in the Russian Federation in accordance with the data of the Federal State Budgetary Institution “Center for Veterinary Medicine”. The species composition of bacteria of the genus Clostridium, which causes diseases in cattle in more than 17 regions of Russia, is described. Moreover, data on the Clostridia species diversity in the Ural region and information on vaccination against blackleg of cattle bred in the Sverdlovsk region are presented. The issues of immunogenicity of vaccines and the effectiveness of their use in cattle, depending on various factors, are highlighted. Up-to-date information on immunobiological preparations against clostridial diseases in cattle officially included in the register of medicinal products of the Russian Federation is presented. The advantages and disadvantages of domestic and foreign vaccines used in veterinary practice are analyzed. A review of current data in the field of new developments of Russian immunobiological agents against clostridial infections in cattle is presented. Comparative data on the use of domestic and foreign vaccines in veterinary practice at agricultural organizations are given. In a brief form, the main preventive measures for each infectious disease caused by clostridia are described.

Veterinary support of the herd horse breeding: problems and solutions

Developed immunobiological preparations for the prevention of infectious diseases in herd horses are presented. Infectious diseases of horses are registered in almost all countries in Asia, Europe and America. In Russia, Kazakhstan, Kyrgyzstan, and Mongolia, the most common diseases are strangles of horses, rhinopneumonia, and salmonella abortion, which cause significant economic damage to herd horse breeding. Various monovaccines are developed and produced in foreign countries, most of them have no use in Russia. We have found a simultaneous disease of mares with rhinopneumonia and salmonellosis, as well as young horses with rhinopneumonia, salmonellosis and strangles. In this regard, the development of monovaccines and combined immunobiological preparations is becoming an urgent problem. Immunogenicity of infectious vaccines should be enhanced by immunomodulators, especially in the extreme conditions of herd horse breeding. At a time when antibiotic-resistant strains of microorganisms are a problem, alternative antibacterial agents should be developed: probiotics and bacteriophages. Eight new microbial strains have been isolated, identified, and deposited in all-Russian collections. Sahabaktisubtil probiotic has been developed, which is used to prevent and treat dysbacteriosis, mycotoxicosis, strangles, leptospirosis, decontamination of manure. New effective vaccine preparations and “Yakutskaya Koumissnaya” starter culture have been developed that can be successfully used to increase the productivity of horse breeding in other regions of the Russian Federation and abroad. Scientific and technical documentation was prepared for inactivated vaccine against rhinopneumonia, combined bivalent and trivalent vaccines. The developed immunobiological preparations are protected by 48 patents for invention and can be used in other countries.

Modern immunobiological preparations for veterinary medicine

With the development of human society, the intensification of agricultural production, climatic changes, environmental pollution, which negatively affect human and animal health, significant changes are taking place in the micro-world. The experience of infectionists show that the most effective way of protection against bacterial, as well as many viral and parasitic diseases, are antibiotics and vaccines.The article provides an overview of immunobiological remedies of veterinary medicine - whole-corpuscular, subunit, genetically engineered, vector and DNA vaccines, vaccines based on virus-like particles and transgenic plants. Whole–body vaccines are live and inactivated biological products, consisting of bacteria or viruses, that retain their integrity during the manufacturing process. Subunit vaccines consist of fragments of infectious disease pathogens capable of providing a specific immune response against a specific pathogen. The technology of manufacturing vector genetically engineered vaccines is based on the use of the virus as a vector for the transfer of genes of protective antigens of other viruses. A gene of the virus of interest is inserted into the genome of the avirulent virus, encoding an antigen that causes an immune response in the vaccinated organism. The avirulent virus modified in this way is used as a live viral vaccine. For the prevention of viral and bacterial diseases in veterinary practice, currently, the advantage is still given to live and inactivated whole-cell biologics, which have a full set of antigens and create intense immunity against infections. The last is the promising direction is the work on the design of genetically engineered vector and DNA vaccines.

Antigenic identity of immunodominant proteins of <i>Bacillus anthracis</i> genovariants

Introduction. The main biological raw materials for the production of immunobiological preparations for identification of Bacillus anthracis are its specific antigens, the protective antigen and the EA1 protein.
 Purpose. To determine the antigenic identity of immunodominant proteins of different genovariants of B. anthracis isolated by gel chromatography and electrophoresis.
 Materials and methods. Culture filtrates of isogenic variants of B. anthracis strain 575/122 (pXO1+, pXO2+): R01 (pXO1+, pXO2); R00 (pXO1, pXO2) were used in the study. Gel chromatographic fractionation and electrophoretic separation were carried out according to standard methods. The antigenic properties of proteins isolated by gel chromatography and electrophoresis were studied by immunodiffusion with polyclonal monospecific sera against the protective antigen and the EA1 protein of the S-layer.
 Results. Gel chromatographic separation of B. anthracis 575/122 culture filtrates R01 (pXO1+, pXO2) and R00 (pXO1, pXO2)yielded fractions 1 and 5. Sera against EA1 protein and antigens of fraction 1 of strains B. anthracis 575/122 R00 and B. anthracis 575/122 R01 culture filtrates identified the identical antigens. Serum against antigens of fraction 5 of B. anthracis 575/122 R01 contained antibodies to numerous proteins, including the protective antigen isolated by electrophoresis.
 Discussion. The antigenic identity of immunodominant proteins isolated by gel chromatography and electrophoresis was identified.
 Conclusion. EA1 and PA proteins isolated by electrophoresis and gel chromatography can be used for production of monoclonal and polyclonal monospecific antibodies suitable for the design of diagnostic preparations.

Development of reassortant influenza vaccines: classical reassortment or reverse genetics?

An important feature of influenza vaccines, which distinguishes them from other immunobiological preparations, is that they have no fixed composition. Due to the constant influenza virus antigenic variability, production facilities require timely supply with relevant vaccine strains undoable due to the lack of proper method for the convenient, rapid and uninterrupted development of vaccine strains. Among the licensed influenza vaccines, classical inactivated and live influenza vaccines hold a special place. They are based on reassortant vaccine strains obtained by crossing currently circulating influenza virus with the so-called donor strain (cold-adapted attenuation donor for live influenza vaccines or high yield donor for inactivated vaccines). Vaccine strains for licensed live attenuated influenza vaccines are reassortants with the so-called 6:2 genome formula two genes encoding hemagglutinin and neuraminidase (HA and NA) belong to the current epidemic virus, and six genes encoding internal proteins (PB2, PB1, PA, NP, M and NS) to cold-adapted master donor virus. There is a very limited number of donors of attenuation. In Russia, there are cold-adapted viruses A/Leningrad/134/17/57 (H2N2) and B/USSR/60/69; in the USA (MedImmune) there are viruses A/Ann Arbor/6/60ca (H2N2) and B/Ann Arbor/1/66ca. MedImmune produces vaccine strains using reverse genetics technique. For other countries, this approach for obtaining vaccines is limited due to the need to purchase a license from the patent holders. In Russia, genetic manipulations with strains for the seasonal live influenza vaccine are not yet allowed; reassortants for the Russian live influenza vaccine are created only by classical reassortment in embryonated chicken eggs. Vaccine candidates for the inactivated influenza vaccine are prepared by the classical reassortment method, the requirements for them are more flexible and allow to use diverse genes combinations from wild type virus and master donor virus. High-yielding viruses such as A/PR/8/34 (H1N1), A/Texas/1/77 (H3N2), B/Lee/40 and some others are used as donors of internal genes. Unfortunately, the classical reassortment method does not always allow to promptly obtain a reassortant virus with a 6:2 genome formula. This is hindered by a number of reasons, ranging from the unique properties of a certain epidemic virus ending up with the constellation of genes. The reverse genetics method based on plasmids is an alternative approach to create reassortant vaccine strains allowing to reliably and quickly obtain reassortant viruses of a set 6:2 genome formula. However, this method also has certain weaknesses. This review discusses the advantages and disadvantages of development of conventional influenza vaccine candidates by reverse genetics and classical reassortment in developing chick embryos.

Review of a series of clinical observations of the use of "Cell culture for substitution therapy"

Introduction. The necessity to develop new substitutional medical technologies is associated with a high demand for rehabilitative measures of traumatological, surgical, and dental patients and persons with burn disease. The existing clinical practice of using allogeneic and autogenous cells, previously used for these purposes, has not yet been standardized. The aim of the study was to summarize scientific literature data on the clinical effectiveness of cell-based medical biotechnologies for regenerative medicine. Materials and methods. The authors reviewed 29 published scientific sources on the results of clinical trials of the preparation "Cell culture for substitution therapy" in 12 medical institutions of the Ural and Siberian regions of the Russian Federation during the period from 1998 to 2012. Results and discussion. Developed and effectively tested on the basis of medical institutions of Sverdlovsk, Chelyabinsk, Tyumen, Kemerovo regions, the Republic of Sakha (Yakutia) and Khanty-Mansi Autonomous Okrug immunobiological preparation "Cell culture for replacement therapy" had high clinical effectiveness in pathological conditions associated with tissue damage in patients with burns, surgery, trauma and dental profile. The studies demonstrated the key activity of allogeneic fibroblasts in the lesion nidus while reducing the time of extracellular matrix formation during active tissue regeneration. The use of the drug prevented the development of generalized infectious-inflammatory complications, reduced the risk of secondary infections, reduced the period of hospitalization and rehabilitation of patients. Conclusion. Analysis of the literature data on the high efficiency of immunobiological agents for substitution therapy on the example of using the preparation on the basis of allogeneic human fibroblasts, based on modern medical standardization and ethical norms, sets the actual task of their replacement by xenogenic and autologous preparations, which will increase the efficiency and safety of the presented method of treatment, improve the quality, life expectancy and demographic indicators in Russia.

Microbiological control of water quality for the production of biological p

one of the main criteria for the biological safety of immunobiological preparations is their sterility. This article presents the results of the evaluation of two methods of direct seeding and membrane filtration. The results of sterility control of the inactivated vaccine against Covid-19 «QazCovid-in®, series № 0400721, № 0410721, № 0420721 are also presented. Evaluation of the sterility of the tested samples of the three vaccine series showed that after incubation, the nutrient medium remained clean both in direct seeding samples and membrane filtration samples, as well as accounting and evaluation of the obtained research data in accordance with the State Pharmacopoeia of the Republic of Kazakhstan. To assess the sensitivity of two methods for determining sterility, samples of immunobiological preparations were experimentally infected with cultures of test strains of St. aureus, C. albicans and C. sporogenes. As a result, the methods of membrane filtration and direct seeding showed the same sensitivity when detecting yeast and anaerobic organisms in all studied concentrations of test strains (10, 1, 0.1 CFU/ml). And for the detection of aerobic microorganisms, the membrane filtration method turned out to be more sensitive compared to the direct seeding method, which is proved by positive results in all samples of test strains with membrane filtration (3/3 in all concentrations) and negative results when setting the direct seeding method (in concentrations of 1 and 0.1 CFU/ml). Thus, the purpose of this study was to evaluate these two methods used to determine the sterility of immunobiological preparations using two methods: direct seeding and membrane filtration

Preclinical Studies of the Specific Activity of the Live Culture Vaccine VACD6 against Smallpox and other Orthopoxvirus Infections

Relevance. The epidemiological situation in the world is characterized by an increase in the incidence of orthopoxvirus infections in humans and animals. In this regard, it is necessary to develop new safe vaccines against these infections.Aim. Conducting preclinical studies on the specific activity of the live vaccine against smallpox and other orthopoxvirus infections VACΔ6 based on the vaccinia virus (VACV) with six deleted virulence genes are presented.Matherials and methods. The studies were performed in accordance with the requirements of the Guidelines for conducting preclinical studies of drugs (immunobiological preparations), the State Pharmacopoeia XIII and the European Pharmacopoeia 7.0.Results and discussion. The vaccine strain VACΔ6 VACV showed significantly reduced neurovirulence in the model of intracerebral administration to suckling mice, and reduced inflammatorynecrotic activity in the model of intradermal administration to rabbits compared to the classical first­generation smallpox live vaccine approved for use in Russia. Preclinical studies of three series of the finished dosage form of the VAC∆6 vaccine showed its authenticity, thermal stability, non­pyrogenicity and safety. Double intradermal vaccination of rabbits at a dose of 106 PFU/animal, a 100% protective effect was provided against the intranasal infection of rabbits with VACV strain HB­-92 at a dose of 1995 LD50 and a double intradermal vaccination of mice at a dose of 106 PFU/animal, full protection was provided against the intranasal infection of mice with ectromelia virus strain K1 at a dose of 56 LD50.Conclusion. Based on the conducted complex of studies, it can be concluded that the created vaccine of the fourth generation VACΔ6 is safer compared to the live smallpox vaccine of the first generation and is not inferior to it in immunogenic and protective properties.

Унификация классификационных групп вакцинопрепаратов

Vaccines – immunobiodrugs, for active immunization of people and animals for preventive and therapeutic purposes, today have received a multi-vector and complex development of technology and structure. At the moment, in the world and in the Russian Federation, in particular, there is no single, generally recognized classification of vaccines that combines all modern achievements in vaccinology. The literature uses disparate approaches to classification, naming of vaccines, as well as incomplete reduction of classification groups. In the presented work, an analysis of existing classifications was carried out, their improvement based on new knowledge in the field of vaccine preparations, the creation of new drugs based on genetic engineering, the systematization of scientific information in the field of development of immunobiological preparations and open prototypes. This classification is proposed to be used to create a unified classification of vaccines on the territory of the Russian Federation and the member countries of the Eurasian Economic Union and use it to standardize finished products, educational and methodological publications, scientific reaserches and other theoretical materials. The paper presents four classification groups, each of which is composed in such a way that, if necessary, for example, in the event of the emergence of fundamentally new technologies for the production of vaccines, they can be implemented into it without compromising the overall systematics.

ОТРАБОТКА РЕЖИМОВ СТАЦИОНАРНОГО КУЛЬТИВИРОВАНИЯ ШТАММА EHV-1/К ВИРУСА РИНОПНЕВМОНИИ ЛОШАДЕЙ

The purpose of these studies is to develop stationary cultivation regimes to determine the most effective ways of viral antigen accumulation while controlling the accumulation kinetics. The novelty of the conducted research is the use of modern methods for analyzing the kinetics of accumulation of the equine rhinopneumonia virus in various modes. Modern production of immunobiological preparations and diagnostic test systems relies primarily on cost-effectiveness. Traditionally, for the accumulation of virus-containing material, a stationary cultivation method is used, which has a number of advantages, primarily associated with the absence of the need for expensive equipment, consumables, and great opportunities for controlling fungal and bacterial contamination, with low staff qualifications. Virological, microbiological, technological and analytical methods of management and control of stationary cultivation of the EHV-1 strain of equine rhinopneumonia virus were used in the work. As a result of the research, a mathematical model of stationary cultivation of the EHV-1 strain was developed, based on the influence of external and internal factors on the kinetics of virus accumulation, with subsequent implementation of the results into the production process. With a standard production process, the biological activity of the virus was 5.75 lg TCID50/cm3, which, when recalculated, is 500 thousand viral particles per 1 cm3, and when the production process was modified using a mathematical model, the virus activity was raised to 6.5 lg TCID50/cm3 , which, when recalculated, is 3 million viral particles per 1 cm3.

From Monitoring the Commitment to Inoculations among Healthcare Workers during the COVID-19 Pandemic to Overcoming Communication Risks in the Process of Vaccination

Introduction: The epidemic process of the novel coronavirus disease (COVID-19) can be managed only through vaccination. Yet, despite the availability of effective immunobiological preparations, health administrators often face a negative attitude of the population, including healthcare professionals, to vaccination. Objective: To establish commitment of workers of health facilities to vaccination against COVID–19 during different periods of the pandemic using modern online resources and to justify the need to develop effective practices of dealing with communicative risks in the course of vaccination. Materials and methods: The study was conducted in several successive stages corresponding to five waves of the COVID-19 pandemic in the Russian Federation by means of an online survey of 5,304 health workers using Google Forms. We applied sociological and statistical methods of research. Results: We observed that the attitude to vaccination against COVID-19 among the employees of health institutions was ambiguous and changed over time demonstrating a gradual increase in adherence to inoculations. The potential risk group with a negative attitude towards vaccination included female nurses and workers of non-medical specialties with less than five years of work experience. An insufficient level of commitment to coronavirus vaccination among the personnel of health facilities has been also noted in other countries, which is a serious problem on a global scale hindering the victory over this novel infection. Conclusion: We assessed commitment to inoculations against COVID-19 among medical workers using modern online resources and found its inadequate level necessitating effective interventions aimed at awareness raising and training in modern practices of effective communications.