Basic and Applied Sciences: Technology and Immunobiological Products.

The experience in organizing the creation of innovative immunobiological drugs and vaccines is discussed-from laboratory developments to industrial technologies and the registration of finished forms of effective and safe drugs that are in demand in the domestic and foreign markets. The principles of their action rely on the latest global achievements in the field of immunobiology and vaccinology. © Pleiades Publishing, Ltd. 2022, ISSN 1019-3316, Herald of the Russian Academy of Sciences, 2022, Vol. 92, No. 4, pp. 452–455. © Pleiades Publishing, Ltd., 2022.Russian Text

Vaccination is viewed as one of the main methods of ensuring the health of any person of any social group in developed and developing countries. It is universally recognized that vaccines reduce child mortality, increase life expectancy, and favor active longevity. In recent decades, world and domestic immunology has made significant progress in the prevention of infectious diseases through national immunization programs. In total, vaccinations against more than 30 different diseases are administered in the world, which has made it possible to achieve a significant reduction in child and infant mortality and to stop or come close to the complete elimination of epidemic threats from many diseases. Vaccines produced using classical technologies are quite successfully coping with problems that were previously considered a threat to national epidemic security. At the same time, as these threats fade away, other difficulties arise. For example, the negative attitude towards vaccination on the part of large population groups and even some doctors is a problem. The pretext for this is the side effects that do exist in some vaccines. Therefore, it is important to pay special attention to improving vaccine production technologies to reduce their reactogenicity and other side effects.

Improvement of vaccine production technologies continues; scientists are abandoning the use of animals and embryos in favor of culturing viruses in cell media. In addition to avoiding ethical problems, this makes it possible to minimize the number of allergic reactions by reducing the number of foreign proteins in the preparation and to standardize its composition. Research is also moving towards the use of genetically engineered compositions containing not a complete antigen but fragments sufficient to induce antibodies but devoid of the pathogenic properties of the virus as such, which significantly reduces the frequency of side effects. RNA and DNA vaccines are being developed that can potentially solve the problems of safety and efficiency in the production of drugs against especially dangerous and poorly cultivated pathogens. Multicomponent vaccines are widely used, allowing vaccination against several pathogens at once [1].

Virologists and infectious disease specialists are beginning to pay attention to the negative effects, including delayed ones, of those infections that previously paled into insignificance against the background of more severe diseases and were not considered a significant threat, for example, rotaviruses and chicken pox. However, a significant increase in healthcare standards makes it necessary to study these infections in depth and to identify their real impact on morbidity and mortality, including child mortality, as well as the long-term consequences of past diseases and the threat to the adult population that does not have immunity to them, as well as to expand the range of names of national vaccination schedules. For adults, as is known, such infections can pose a danger of a completely different level than for children. Therefore, it is necessary to create and master the production of new vaccines against pathogens that were not previously considered worthy of serious attention and to include them in national calendars. Abroad, such work has long been actively carried out, while the domestic industry dedicates little attention to this problem. Lagging behind in the field of innovative prophylactic and therapeutic immunobiological drugs can lead to the formation of almost complete dependence on foreign manufacturers when using innovative methods of treatment and prevention of mass and socially significant diseases—from infectious to oncological.

Since the number of newly discovered viruses is growing, over time, vaccination will only be possible in the form of complex vaccines against several infectious agents at once. The synthesis of the viral genome makes it possible to guarantee its composition and genetic homogeneity. In addition, there are ways to attenuate (weaken) viruses, which are being implemented using, among other things, genomic modifications.

Over the past few decades, vaccine development technologies have undergone significant changes owing to a better understanding of the functioning of the mechanisms of the human immune system in the fight against infectious agents and malignant tumors. Along with preventive vaccines, the effectiveness and safety of which are significantly increasing due to the use of new technologies, therapeutic vaccines have appeared and are increasingly being used. Based on knowledge about the mechanisms of interaction between the immune system and viruses, gene therapy preparations have appeared that use viruses as vectors.

Immunobiology is beginning to be used in oncology. A trend that uses the vulnerability of a number of cancer cells to the virus is actively being developed [2]. The treatment technology consists in the selection of a minimally pathogenic viral strain (for example, reovirus, rotavirus, modified pox virus, Coxsackie virus) for use as an agent that causes lysis (destruction) of the tumor. Scientific work is being carried out in the direction of searching for mechanisms and identifying patterns of vulnerability of tumors of various etiologies and selecting strains and methods for using oncolytic viruses, as well as technologies for their industrial production. The first trials have shown that in some cases the survival rate of patients without tumor progression (when treatment is suspended) is doubled, and overall survival in some patients can be increased by 3‒4 times [3].

The Chumakov Federal Science Center for Research and Development of Immunobiological Products, RAS, is a leading scientific center in the field of medical virology, including the study of poliomyelitis; tick-borne encephalitis; viral hemorrhagic fevers; influenza; and enterovirus, arbovirus, and coronavirus infections. It conducts basic and applied, including clinical, research and develops the scientific basis for the creation of preventive and diagnostic medicines, and theoretically substantiates strategies for the prevention of infectious diseases and studies issues of ensuring biosafety of the environment. The intellectual and technical potential of the center allows it to carry out a full cycle of work (from the creation of the concept of a drug and laboratory research to the technology for manufacturing finished dosage forms) and to organize and control the necessary volume of preclinical and clinical studies, as well as the process of registration of medicines, including in foreign markets.

The main directions of basic research of the Chumakov Center are the following:

• study of the biology of viruses that cause infectious diseases of high social significance and their interaction with the host at the molecular, cellular, organismal, and population levels [4];

• study of the mechanisms of variability of RNA-containing viruses based on the analysis of materials from patients, animals, and environmental objects, as well as experimental model studies [5];

• experimental study of virus‒cell interaction and the molecular basis of pathogenesis and formation of an immune response in viral diseases [6];

• study of the factors that determine the epidemiological and epizootological situation for viral infections with different ways of spread and the development of a scientifically justified scenario for changing the situation depending on external influences [7, 8];

• study of the structural and functional organization of parasitic systems in natural foci of new and recurring infections, as well as the mechanisms of bringing pathogens into the territory of Russia and preventing their spread [9].

Technological solutions for applied research problems of the Chumakov Center include the following:

• development of biotechnological fundamentals for the creation of immunobiological preparations, including isolation and identification of viruses (using physicochemical, biological, immunological, molecular, and electron microscopic methods), study of the spectrum of cell cultures sensitive to virus propagation, certification of vaccine strains of viruses, determination of the range of laboratory animals as a biological model for studying the clinical and immunological manifestations of infection, optimization of virus cultivation to obtain a highly active substrate, filtration of virus-containing liquid, concentration of virus-containing liquid (ultrafiltration in tangential flow), purification of the virus-containing concentrate (gel chromatography), and inactivation with formalin or beta-propiolactone;

• development of vaccine quality control methods;

• preclinical studies of the vaccine;

• epidemiological monitoring of known and emerging infections.

The scheme of the biotechnological platform for whole-virion vaccines is shown in Fig. 1. Vaccine preparations produced at the Chumakov Center for the prevention of viral diseases are shown in Fig. 2. The main products of the center are antiviral vaccines against rabies, tick-borne encephalitis, and yellow fever, as well as the oral polio vaccines created over the past five years, including the monovalent drug MonoVacPolio (based on attenuated Sabin poliovirus 1, 2, 3 serotypes) and the bivalent drug BiVacPolio (based on Sabin poliovirus 1 and 3 serotypes). In addition, for the first time in Russia, an inactivated polio vaccine from attenuated Sabin viruses of three serotypes, PoliovacSin, has been registered and is ready for industrial production.

Fig. 1.

Biotechnology platform for whole virion vaccines.

Fig. 2.

Vaccine preparations produced in the Chumakov FSC R&D IBP, RAS.

In connection with the COVID-19 infection pandemic, a manufacturing technology for the inactivated whole-virion vaccine CoviVac was developed and its large-scale production was mastered [10]. By now, the bivalent whole-virion inactivated vaccine HFRS-Vac for the prevention of hemorrhagic fever with renal syndrome, which has no analogues in the world, has successfully passed preclinical studies [11].

The prospects for technological solutions in the production of vaccines at the Chumakov Center are the following.

• Development of an inactivated coronavirus vaccine based on new strains: CoviVac-Delta, CoviVac-Combi, and a Combined covid and influenza vaccine.

• A vaccine against poliomyelitis based on viruslike particles in plants. For the development of this technology, the genes of poliovirus capsid proteins have been obtained and modified. Genetically engineered constructs based on various vectors have been created for the expression of viruslike particles of poliovirus in plants. Methods have been developed for delivering the constructs to producer cells, as well as the main methods for isolating and detecting viruslike particles in plants and methods for growing plants under the conditions of an aeroponic installation for the expression of viruslike particles. In the near future, the expression of viral proteins and viruslike particles in plants under phytotron conditions will be worked out, and the efficiency of their expression under various conditions will be analyzed.

• A vaccine against COVID-19 based on viruslike particles in insect cells, for the development of the technology of which capsid protein genes have been isolated from the AYDAR-1 vaccine strain. Donor plasmids and recombinant bacmids for the expression of capsid proteins have been obtained. Recombinant baculoviruses carrying the S and N genes have been obtained. The technology for cultivating Sf9 insect cells on a laboratory scale has been developed. The envelope proteins of SARS-CoV-2 Spike and N have been expressed. In the near future, recombinant baculoviruses carrying the SARS-CoV-2 M and E genes will be obtained. The conditions for coexpression of the SARS-CoV-2 envelope genes to obtain viruslike particles will be selected, and the technology of purifying viruslike particles will be developed.

Considering new knowledge, biotechnologists should rely on the development of innovative vaccines based on the latest achievements in vaccinology using advanced systems and approaches that combine areas such as genomics, transcriptomics, proteomics, etc. It is necessary to move actively from traditional methods of creating vaccines to modern ones, aimed at enhancing the stability of vaccines and improving their composition and methods of delivery. Lack of attention to this trend can lead (and is already leading) to a gradual “washout” of the domestic range of vaccines by Western analogues produced using more modern technologies, which, among other things, are promoted based on the opinion of the WHO.