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Objective of the study was to analyze the stability of the preparation of a live plague vaccine made using an experimental nutrient medium over its shelf life. Materials and methods . A series of live plague vaccine based on Yersinia pestis EV NIIEG strain prodused using experimental nutrient medium were utilized in this work. Most significant qualitative parameters of the preparation were studied: microbe cell content, viability, thermal stability, mass loss during the process of drying, and immunogenicity. Results and discussion . The stability of the vaccine produced using the nutrient medium based on hydrolysate of condensed corn steep extract was monitored over a specified shelf life (three years). A comparative analysis of the viability of all experimental vaccine series was carried out at certain time intervals - short-term storage (up to 3 months) and before the expiration date. During storage, there was an insignificant decrease in the percentage of living microbial cells, while in no series the viability decrease reached below the regulated levels of 25 %. Such a decrease is natural for a “live preparation” and is not critical within the obtained limits. The rest of the studied indicators practically did not change. Thus, the analysis of the data obtained indicates that in all the periods of observation the drug retained the stability of the main regulated indicators. The studies confirm the feasibility of using this referred nutrient medium based on hydrolyzed corn steep extract in industrial production of the plague live vaccine preparation.

Objectiveof the study was to test the nutrient medium based on the enzymatic hydrolysate of corn extract condensed for a scaled production of live plague vaccine and to check the quality of the obtained batches against the specified parameters.Materials and methods.A dense nutrient medium based on corn extract was used to grow biomass in the process of live plague vaccine production. The quality parameters of the vaccine preparation obtained were studied by the regulated methods set forth in the regulatory documentation.Results and conclusions. The vaccine was monitored at all stages of its manufacture, including control of the finished dosage form, in strict accordance with the approved regulatory documentation. All the experimental production series complied with the specified indices. Approbation of the production cycle environment for live plague vaccine manufacturing showed efficiency of the conditions and the possibility of environment’s application in the industrial production of the preparation.

Objective of the study is to specify the character and dynamics of morpho-functional changes in the organs of peripheral immune system of BALB/c mice in case of immunization with vaccine Yersinia pestis EV NIIEG strain against the background of immune modulation. Materials and methods. Applying a combination of histological, histo-chemical, immune histo-chemical methods, and morphometric analysis, defined has been the targeting of immunological processes in peripheral lymphoid organs of BALB/c mice. Results and conclusions. Revealed has been stimulating effect of polyoxidonium in concomitant use of Yersinia pestis EV NIIEG, on the processes of cell proliferation in T-zones of lymphoid organs, on activation of T- and B-lymphocyte sub-population in them by reference to the increment in the amount of specifically stained with CD25 marker cells and nuclei of lymphocytes with high argyrophil granule content, indirectly characterizing the functional state of nuclei apparatus, as well as on response intensification of neuroendocrine environment of lymphoid structures. Based on the results of complex morphological analysis, specified have been the peculiarities of morphofunctional changes in immune competent organs of biomodels, being indicative of polyoxidonium potential to increase the effectiveness of vaccination against plague.

Objective of the study is to develop a reagent for the detection of lymphocytes with Yersinia pestis F1 antigen receptors. Materials and methods. Utilized have been: live plague vaccine based on the strain of Yersinia pestis EV NIIEG, formalin killed suspensions of microorganisms - Y. pestis , 3123, Y. enterocolitica O9 H-383 serovar, Y. pseudotuberculosis O1 2841 serovar; acetaldehyde-immobilized capsular antigen of Y. pestis F1 (obtained applying Baker methodology), lipopolysaccharide of Y. pestis K1, and bovine erythrocytes. Bovine erythrocyte F1 sensibilization has been performed using rivanol. Lymphocytes from blood have been isolated in density gradient ficoll-verografin 1.077. Lymphocytes with Yersinia pestis F1 antigen receptors have been detected by means of reagent adhesion onto the isolated lymphocytes. F1-free erythrocytes serve as controls. After the exposition, 7 evaluations of specificity to F1 and the lymphocytes, binding control reagent, have been carried out. Deployed have been 8 rabbits, immunized with live vaccine EV, and 2 rabbits, immunized with inactivated vaccine EV. Examined have been EV-vaccinated 5 persons. Results and conclusions. Identified is optimum sensibilizing dose of F1 antigen (250 µg/ml). Specificity of lymphocytes with receptors to F1 is demonstrated in inhibition experiments applying homologous and heterogeneous antigens. Lymphocytes with receptors to F1 (LRs) have been detected in peripheral blood of all rabbits and humans, immunized with vaccine EV. LRs have been registered since day 2 till day 35 in the rabbits, and in humans - since day 2 till day 14 after vaccination. It is shown that in case of revaccination of humans, LRs emerge and disappear earlier, than in case of primary immunization.

Nowadays, problem of tableted drug form contamination with extraneous micro-flora is in the spotlight of scientists, as the specific share of these medical preparations in the world market amounts to more than 60 % and has a tendency to increase. Thus, objective of the study is to investigate the degree of contamination of the basic and auxiliary raw materials at different stages of live plague vaccine manufacturing, rapid dissolving tablets, and the ways to reduce it. Materials and methods. Utilized has been lyophilized Y. pestis live culture of the vaccine strain EV NIIEG, and the live plague vaccine, rapid dissolving tablets. Carried out has been assessment of “microbiological purity” at different stages of tableted live plague vaccine manufacturing: grinding, mixing, granulation, sublimation, and palletizing. Results and conclusions. Identified is the dynamic pattern of quantitative micro-flora composition of the mentioned above drug. Established is the alteration of microbial impurity at separate technological manufacturing steps. Specified is the technological stage with the most expressed contamination. Analysis of factors, which affect vaccine impurity, has revealed that finished dosage-form quality improvement is impossible without incoming control of stock and auxiliary materials, as well as enhancement of manufacturing procedure up to the level, complying with applicable pharmaceutical production standards. It is experimentally proved that series of tableted live plague vaccine, obtained using modernized technological equipment, provide for 3-fold reduction of contamination.

The essential index of functional activity of innate immunity cells in inflammatory reactions, induced by specific allergens, is the secretory neutrophils degranulation with the release of serine proteases from granules on the cells' surface and into the extracellular space. Using flow cytofluorimetry blood granulocytes of persons, immunized with live plague vaccine, were shown to respond to in vitro contact with allergen of plague microbe (pestin) by degranulation, in contrast to those of unvaccinated people. The degree of the innate immunity cells activation, as regards this index, depends on the time period passed after the vaccination. The received experimental data agree with modern conceptions on the leukocytic proteases` role in the development of allergic inflammation of skin. They also provide the necessary basis for further investigations, required to develop rapid instrumental test for human anti-plague immunity assessment in vitro .

Arabinoganactane isolated from Siberian larch (Larix sibirica) and its iron and cobalt nanobiocomposites were selected as immunomodulators to elucidate the possibility of intensification of specific immunity against plague. Comparative morphometric analysis of immune system organs of white mice inoculated with live plague vaccine (LPV) and its combination with immunomodulators was performed. Experimental data indicate that arabinogalactane and its nanobiocomposites used in combination with LPV increase immunocytopoetic function of immune system organs to larger extent than LPV.

The preference of the (21±1) °C temperature for cultivation of the vaccine strain Yersinia pestis EV in the live plague vaccine biomass production to (27±1) °C is experimentally confirmed. An original interpretation of the mechanism of small cells stability that are formed at (21±1) °C has been proposed. This interpretation stipulates a smaller damaging effect of steam molecules during sublimation in the process of lyophilization.