Medical academic journal最新文献

{"title":"Cellular immune response to combined DNA-protein constructs carrying SARS-CoV-2 antigens","authors":"A. M. Zadorozhny, M. Borgoyakova, E. Starostina, L. Karpenko","doi":"10.17816/maj108654","DOIUrl":"https://doi.org/10.17816/maj108654","url":null,"abstract":"BACKGROUND: One approach that makes it possible to create vaccines relatively quickly and inexpensively is the creation of DNA vaccine constructs. They are of particular interest for the prevention of COVID-19, as they induce both types of immune response humoral and T cell. Previously, we have created two DNA vaccines based on the pVAX vector: one encodes the full gene of the S protein of the SARS-CoV-2 virus, the second only encodes the receptor-binding domain (RBD) of the S protein of the SARS-CoV-2 virus. Next, the naked DNA was wrapped in a polycationic polyglucin-spermidine complex conjugated with the RBD protein. The resulting combined DNA-protein constructs were named CCV-RBD and CCV-S, respectively. \u0000AIM: To investigate the induction of the T cell immune response of the developed combined DNA protein candidate vaccines in an animal model. \u0000MATERIALS AND METHODS: BALB/c mice were immunized with constructs CCV-RBD and CCV-S, after which their spleens were removed from which splenocytes were isolated. The cellular response was assessed by the ability of splenocytes to secrete cytokines in response to stimulation with viral peptides. The intensity of the response was recorded using the intracellular cytokine staining (ICS) method using flow cytometry. \u0000RESULTS: BALB/c mice were immunized twice with an interval of three weeks with a dose of 100 g of DNA (8 animals per group): 1) CCV-RBD 2) CCV-S and 3) intact animals. It has been shown that both T helper lymphocytes (CD4+) and cytotoxic lymphocytes (CD8+) of animals immunized with CCV-S and CCV-RBD respond with the release of cytokines in response to stimulation with viral peptides. \u0000CONCLUSIONS: In the case of CCV-RBD, a trend towards a higher response in both CD4+ and CD8+ was observed compared to the CCV-S group. Possibly, this difference may be due to more efficient synthesis of the RBD protein than the S protein, providing a DNA vaccine.","PeriodicalId":342669,"journal":{"name":"Medical academic journal","volume":"44 10 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122069053","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Method of obtaining purified SARS-CoV-2 antigen for structural and immunological studies in whole-virion vaccines against coronavirus infection","authors":"A. Zyrina, Irina O. Tselykh, Anastasia S. Bogdan, A. Kovpak, Y. Ivin, V. E. Vasilenko, A. Piniaeva, А.А. Shishova","doi":"10.17816/maj108681","DOIUrl":"https://doi.org/10.17816/maj108681","url":null,"abstract":"BACKGROUND: Despite the large-scale development of vaccines against coronavirus infection, there is still no complete information about the antigenic structure of the SARS-CoV-2 virus particles. This article describes a method of obtaining a pure concentrated whole-virion sample that can be used in various studies. \u0000AIM: The goal is to develop an optimal method of purification of the inactivated SARS-CoV-2 virus in order to obtain a standard with parameters of purity and antigen content sufficient for structural and immunological studies. \u0000MATERIALS AND METHODS: To obtain a pure concentrated virus SARS-CoV-2 we used the sucrose density gradient ultracentrifugation. Fractions with the highest content of viral particles were evaluated based on the concentration of nucleocapsid N and glycoprotein S. The purity of the pooled fractions (the purified sample) was evaluated by the presence of impurity proteins, toxins and bovine serum albumin. \u0000RESULTS: The optimal conditions for obtaining the inactivated purified SARS-CoV-2 whole virus were determined. A standard sample of the inactivated SARS-CoV-2 virus was isolated and characterized. \u0000CONCLUSIONS: The obtained standard sample can be used in enzyme immunoassay to measure the amount of antigen in whole-virion vaccines against coronavirus infection, as well as in various structural studies of SARS-CoV-2 virus particle.","PeriodicalId":342669,"journal":{"name":"Medical academic journal","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122123282","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Role of the intestinal microbiota in the pathogenesis of multiple sclerosis. Part 1. Clinical and experimental evidence for the involvement of the gut microbiota in the development of multiple sclerosis","authors":"I. Abdurasulova","doi":"10.17816/maj108241","DOIUrl":"https://doi.org/10.17816/maj108241","url":null,"abstract":"The review discusses the complex role of the intestinal microbiota in the pathogenesis of multiple sclerosis, summarizes data from studies of changes in the composition of the intestinal microbiome in patients with multiple sclerosis, and provides evidence of the involvement of the intestinal microbiota in the development of experimental autoimmune encephalomyelitis in animals, a valid model of multiple sclerosis.","PeriodicalId":342669,"journal":{"name":"Medical academic journal","volume":"27 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128577370","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modulation of porcine alphacoronavirus pathogenesis by antibodies to SARS-CoV-2","authors":"Mariya V. Nefedeva, I. Titov, S. Katorkin, S. Tsybanov, V. Lyska, A. Sereda, A. Malogolovkin","doi":"10.17816/maj108729","DOIUrl":"https://doi.org/10.17816/maj108729","url":null,"abstract":"BACKGROUND: A variety of antibodies against the SARS-CoV-2 with different functional activities can lead to immune pathology and/or modulation of infection due to antibody-dependent enhancement. An important role in the development of ADE, as exemplified by Dengue fever, is played by low-affinity antibodies, which interacting with the virus, do not neutralize it, but on the contrary forms the virus-antibody complex to immune cells (monocytes/macrophages, B-cells). There are conflicting data on the antibody-dependent enhancement presence in COVID-19 coronavirus infection. Some studies indicate a relationship between the severity of infection in patients infected by SARS-СoV-2 and the level of antibodies to closely related coronaviruses. \u0000AIM: The aim of the study was to understand the possibility of interaction between antibodies to SARS-CoV-2 with closely related porcine coronavirus (alphacoronavirus) and their ability to modulate the development of infection in vitro. \u0000MATERIALS AND METHODS: In the study serums and leukocytes from patients, who are convalescent after COVID-19 infection in 2020, was used. Antibody titers (IgG/IgM) was checked by commercial kit (VektorBest, Russia), based on ELISA method. In the eukaryotic expression system Chinese hamster ovary cells, monoclonal antibody to SARS-CoV-2 spike protein and its Fab version without Fc domain were obtained. As a model, swine alphacoronavirus- transmissible gastroenteritis virus (TGEV) was used. Replication activity of TGEV was studied in porcine leukocyte cell culture. The ability of this virusto induce the interferon gamma synthesis in leukocytes from convalescents was assessed TigraTest SARS-CoV-2 Enzyme-Linked SPOT analysis (Russia). Virus titration and neutralization assay with sera and monoclonal antibodies were carried out in porcine kidney PK-15 cell line. \u0000RESULTS: Serum samples from 43 donors (59% female, 41% male) aged from 21 to 56 was studied for the presence of class M and G antibodies to SARS-CoV-2 by ELISA method. Class M antibodies were found in only one donor. The highest titers of class G antibodies (1:400 to 1:1600) were detected among donors 4050 years old. For four donors with the highest antibody titers, HLA-II type was determined. The DQA1 allele in 3 patients was found to have the *0501 variant, which, according to literature data, is associated with autoimmune thyroid diseases. For the DQB1 allele, two convalescents had exactly the same variants (*0602-8). Among HLA DRB1, all patients had different allele variants (*09, *11, *01,*03,*13,*15,*08,*13). \u0000Among the samples, studied by ELISPOT method, in 45,5% of samples a positive T-cell response was noted after stimulation of macrophages with the S peptides of the SARS-CoV-2 protein and in 22.7% of samples in response to the pool of N peptides of the SARS-CoV-2 protein. Interestingly, infection of macrophages from convalescents with the transmissible gastroenteritis virus caused interferon-gamma expression in 31,8%","PeriodicalId":342669,"journal":{"name":"Medical academic journal","volume":"58 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123134701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Activation and modulation of Ire1-Xbp1 pathway in SARS-CoV-2 infected Vero cells (B.1.1.7 Alpha, B.1.617.2 Delta, B.1.1.529 Omicron variants)","authors":"A. Shishova, V. Baryshnikova, Maya Yu. Ermakova, Yuriy Turchenko, Alena Dereventsova, Kseniya V. Fominykh","doi":"10.17816/maj108694","DOIUrl":"https://doi.org/10.17816/maj108694","url":null,"abstract":"BACKGROUND: The emergence of new variants of the SARS-CoV-2 virus at the end of 2020, which became a source of increased risk for global public health, prompted the study of their molecular biological characteristics and pathogenic effects. It is known that one of the reasons for the viruses pathogenic action is their interaction with the defense mechanisms of the cell. Ire1 (inositol-requiring enzyme 1)-mediated splicing of Xbp1 mRNA (X-box binding protein 1) is a protective mechanism that is activated in response to the accumulation of misfolded proteins in the cell, a situation that occurs due to uncontrolled synthesis of viral proteins during infection. Studying the interaction of different variants of the SARS-CoV-2 virus with this protective mechanism will help to shed light on various aspects of the pathogenesis of a new coronavirus infection. \u0000AIM: Study of Ire1-Xbp1 defense mechanism activation and modulation in SARS-CoV-2 infected Vero cells. \u0000MATERIALS AND METHODS: We studied the activation of the Ire1 enzyme in Vero cells infected with various variants of the SARS-CoV-2 virus using immunoblotting and antibodies to various forms of this protein in the cell. In addition, we studied the activation of Xbp1 mRNA splicing under conditions of infection with various variants of the SARS-CoV-2 virus in a PCR reaction with specific primers. \u0000RESULTS: Reproduction of B.1.1.529 (Omicron) strain in Vero cells is slower than B.1.1.7 (Alpha) and B.1.617.2 (Delta) strains of SARS-CoV-2. The whole reproduction cycle is 48 hours. \u0000Ire1-dependent defense mechanism is activated after 12 hours of SARS-CoV-2 infection with either of three variants. However, despite the activation of the Ire1 endonuclease domain, there is no Xbp1 mRNA splicing in SARS-CoV-2 infected cells. Inhibition of Xbp1 mRNA splicing occurs slower in Vero cells infected with the B.1.1.529 Omicron variant. \u0000CONCLUSIONS: The paper describes the reproduction of various variants of the SARS-CoV-2 virus in Vero cell culture and the activation of the Ire1-Xbp1 defense mechanism during infection. The Ire1 endonuclease is phosphorylated, however, mRNA splicing of the Xbp1 transcription factor is impaired in SARS-CoV-2 infected cells. A decrease in the rate of inhibition of this protective mechanism in Vero cells infected with the Omicron (B.1.1.529) variant of the SARS-CoV-2 virus may be the reason for its lower pathogenicity described in various studies.","PeriodicalId":342669,"journal":{"name":"Medical academic journal","volume":"14 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115431440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Аttenuation мarkers of cold-adapted SARS-CoV-2 variants","authors":"A. Gracheva, E. Korchevaya, R. Samoilikov, D. Smirnova, I. Leneva, A. Poromov, A. Pankratov, G. Trunova, V. Khokhlova, F. G. Nagieva, O. A. Svitich, V. Zverev, E. Faizuloev","doi":"10.17816/maj108725","DOIUrl":"https://doi.org/10.17816/maj108725","url":null,"abstract":"BACKGROUND: Unprecedented anti-epidemic measures and the widespread use of vaccines against COVID-19 have reduced the rate of hospitalization and mortality from the disease, but have not stopped the SARS-CoV-2 pandemic spread. The development of live vaccines against COVID-19, capable of providing the formation of a long-term humoral and cellular immune response and cross-protection against new SARS-CoV-2 variants of concern, is relevant. Previously at the I.I. Mechnikov Research Institute of Vaccines and Sera SARS-CoV-2 cold-adapted (ca, cold-adapted) variants were obtained. This work is aimed to search for methodological approaches that allow in vitro screening studies to assess the attenuation (att) phenotype of ca SARS-CoV-2 variants. \u0000MATERIALS AND METHODS: The SARS-CoV-2 laboratory strain Dubrovka and its variants were cultured in Vero and Calu-3 cells. Quantitation of the virus was carried out by titration in Vero cells and by real-time RT-PCR. The attenuation (att) phenotype of SARS-CoV-2 variants was determined on an animal model of COVID-19 on Syrian hamsters. \u0000RESULTS: In experiments on Syrian hamsters, the presence of the att phenotype in the ca variants of the virus was established. Animals infected with virus ca variants had significantly less weight lost, had less viral load in the lungs and brain and less pronounced pathological changes in the lungs compared to infection with the virulent strain. In vitro experiments on Vero and Calu-3 cells revealed probable attenuation markers of the virus ca variants for syrian hamsters: (1) ability to reproduce at low temperature (ca phenotype); (2) inability to reproduce at 39 C (ts phenotype); (3) changes in the species and tissue specificity of the virus. \u0000CONCLUSIONS: The developed methodological approaches to the identification of SARS-CoV-2 attenuation markers are a valuable tool for monitoring the stability of the phenotype of candidate vaccine strains.","PeriodicalId":342669,"journal":{"name":"Medical academic journal","volume":"10 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126572568","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ways of spread, detection, clinical course of syphilis and leprosy among the population of the north-eastern districts of Eastern Siberia in the 19th century","authors":"G. Gaidarov, T. I. Alekseevskaya, T. Demidova, O. Sofronov","doi":"10.17816/maj108403","DOIUrl":"https://doi.org/10.17816/maj108403","url":null,"abstract":"BACKGROUND: In the 19th century, the process of annexation of lands in the north-east of the Russian Empire was completed. The influx of newcomers, political exile, criminal hard labor created the prerequisites for an aggravation of the epidemiological situation in the newly annexed territories. The morbidity of the population began to acquire an epidemic character. Syphilis, which spread in the 19th century, leprosy struck foreign communities in the northern and eastern regions of the Pacific Ocean, the Sea of Okhotsk, the Kamchatka Peninsula and the alien population from other territories. \u0000AIM: Systematize, as well as identify certain trends in the prevalence and forms of manifestation of syphilis and leprosy among the population of the north-eastern districts of Eastern Siberia of the 19th century. according to the reporting materials of doctors and authorities of that time. Let us explain: in this article, the authors quote from the documents, preserving (if possible) the style and punctuation of the originals, and also use geographical names and names of territories adopted in the Russian Empire in the 19th century. \u0000MATERIALS AND METHODS: On the basis of archival materials, the article discusses the ways of importation and spread of syphilis among representatives of individual foreign communities, the clinical course and forms of manifestation. The multiplicity of ways of penetration of syphilis to the outskirts of Eastern Siberia is noted. The reports of seconded staff physicians, doctors, paramedics during the examination of the population and the treatment of patients in hospitals provide the first statistical information on the incidence of men, women, their nationality, and the outcome of treatment. The reports of seconded staff physicians, doctors, paramedics during the examination of the population and the treatment of patients in hospitals provide the first statistical information on the incidence of men, women, their nationality, and the outcome of treatment. \u0000RESULTS: The article highlights the issues of studying the role of factors contributing to a high level of the spread of syphilis and leprosy among inorodtsy (indigenous dwellers), describes the symptoms of manifestation, and especially chronic syphilis, and notes the role of unsanitary living conditions and household arrangements in the spread of diseases. \u0000CONCLUSIONS: Observation of the clinical manifestations of syphilis allowed doctors to isolate leprosy as an independent form of the disease, and to investigate its causes. These observations were the basis for the development of measures aimed at stopping the spread of the disease among the population. The beginning of the emergence of medical and police supervision aimed at identifying patients, registering them and treating them is noted.","PeriodicalId":342669,"journal":{"name":"Medical academic journal","volume":"17 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123921458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of the course of the new coronavirus infection COVID-19 in pregnant women under treatment in the Infection Department of Clinical Hospital no. 1 in Smolensk","authors":"Anastasia A. Skvortsova, A. Zakharova, Sofia A. Kostina, Olga S. Rozinkova, Anna B. Melnikova, E. Simakina","doi":"10.17816/maj108625","DOIUrl":"https://doi.org/10.17816/maj108625","url":null,"abstract":"BACKGROUND: The physiological features of the pregnant womans body include her in the risk group for a more severe course of the disease, therefore, the management and treatment of such patients requires the most careful monitoring, as well as the coordinated work of a gynecologist and an infectious disease specialist. \u0000AIM: To evaluate the variants of the course of the disease in pregnant women with confirmed coronavirus infection. \u0000MATERIALS AND METHODS: The medical records of patients who were treated in the infectious diseases department of the Clinical Hospital No. 1 in Smolensk from May 2020 to September 2021 were analyzed, the necessary data were included in the protocols of our study. Such indicators as the main diagnosis and severity of the course of the disease, complaints at admission, the presence and stage of fever, age, gestational age, the presence of concomitant pathology and a complicated obstetric and gynecological history were evaluated. \u0000RESULTS: Analysis of the course of the disease according to the medical history showed that a moderate form of a new coronavirus infection dominates pregnant women. \u0000CONCLUSIONS: A trend towards a moderate course of a new coronavirus infection COVID-19 has been established. The probability that this is due to the presence in most patients with a history of diseases of the cardiovascular system, kidneys, liver, as well as diabetes, obesity, anemia.","PeriodicalId":342669,"journal":{"name":"Medical academic journal","volume":"31 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121626378","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessment of cell-mediated immune responses to SARS-CoV-2 in Syrian hamsters","authors":"D. Mezhenskaya, I. Isakova-Sivak, L. Rudenko","doi":"10.17816/maj108629","DOIUrl":"https://doi.org/10.17816/maj108629","url":null,"abstract":"BACKGROUND: The pandemic caused by the SARS-CoV-2 virus at the end of 2019 remains to be a serious healthcare problem. Constant antigenic drift of the pathogen led to a decrease of licensed COVID-19 vaccines effectiveness. And the development of broad-spectrum vaccines with high effectiveness rate against evolutionarily divergent SARS-CoV-2 variants remains an urgent issue. Unlike virus-specific antibodies with limited spectrum of action, T-cell immunity has a wider cross-protective potential. Syrian hamsters are the most appropriate model for preclinical evaluation of new vaccine candidates, since these animals are susceptible to SARS-CoV-2 infection and show clinical symptoms of the disease. However, study of T-cell vaccine response in hamsters is complicated by the lack of available reagents and test systems for adequate assessment of the virus-specific cellular immunity levels after vaccination. \u0000AIM: In this work, we report an optimized protocol of stimulation of Syrian hamsters immune cells with a live SARS-CoV-2 virus to assess virus-specific T-cell responses. \u0000MATERIALS AND METHODS: Intranasal infection of animals with SARS-CoV-2 virus followed by stimulation of immune cells with different doses of whole live coronavirus and counting of IFN-producing cells by ELISpot method. \u0000RESULTS: Stimulation of spleen and lung cells with SARS-CoV-2 at a dose 0.1 TCID50/cell is the most optimal viral concentration for detecting maximum of cytokine-producing cells in SARS-CoV-2-infected animals. Stimulation of cells with whole virus revealed greater number of virus-specific cells compared to a stimulation with pools of SARS-CoV-2 lyophilized peptides (S and N proteins). \u0000CONCLUSIONS: Overall, the new methodology allows assessment of the immunogenicity of COVID-19 T-cell vaccines more accurately in preclinical studies using the Syrian hamsters model.","PeriodicalId":342669,"journal":{"name":"Medical academic journal","volume":"6 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116602900","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a biosafe ELISA-based platform for assessing immunogenicity in the production of an inactivated whole-virion coronavirus vaccine","authors":"Dmitry V. Danilov, O. A. Shmeleva, A. S. Lunin, L. Kozlovskaya, A. Piniaeva, A. Shishova","doi":"10.17816/maj108717","DOIUrl":"https://doi.org/10.17816/maj108717","url":null,"abstract":"BACKGROUND: SARS-CoV-2 vaccine immunogenicity is evaluated in neutralization test with live virus. It is performed in a biosafety level 3 zone because requires live virus stage. Therefore, control laboratories should be certified for this class of work. The development of technology based on enzyme-linked immunosorbent assay as an analogue of the neutralization reaction makes it possible to create an immunobiological product in a shorter time and in conditions without special requirements for control laboratories. \u0000AIM: Development of an enzyme-linked immunosorbent assay for assessing SARS-CoV-2 vaccine immunogenicity by measuring neutralizing antibodies production in immunized animals. \u0000MATERIALS AND METHODS: Recombinant receptor-binding domain fused to a С-terminal hexahistidine sequence was produced in Escherichia coli cells and purified via metal-affinity chromatography on WorkBeads NiMAC (Bio-Works). Purified protein was used in enzyme-linked immunosorbent assay as an antigen for sorption. The sera of mice immunized with the vaccine preparation were tested for neutralizing activity against the SARS-CoV-2, as well as in the developed enzyme-linked immunosorbent assay. \u0000RESULTS: Sera with high neutralizing titers showed a high degree of binding to recombinant receptor-binding domain fused to a С-terminal hexahistidine sequence in enzyme-linked immunosorbent assay, while sera from non-immunized animals or sera with neutralization titers less than 1:8 were not reactive in enzyme-linked immunosorbent assay. The Spearman and Pearson correlation coefficients for neutralization test titers and optical density in enzyme-linked immunosorbent assay were 0.759 and 0.76, respectively. The developed assay can be used as a semi-quantitative method for assessing the immunogenicity of a vaccine against coronavirus infection. \u0000CONCLUSIONS: The developed platform makes it possible to reliably assess the immunogenicity of an inactivated coronavirus vaccine under conditions that do not require a high biosafety conditions.","PeriodicalId":342669,"journal":{"name":"Medical academic journal","volume":"40 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2022-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127124232","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}