Polina P. Goldovskaya

Filoviruses cause highly lethal infectious diseases with hemorrhagic symptoms and extremely high fatality rate (up to 90%). Although two vaccines against Ebola virus (EBOV) are licensed for application in endemic areas, vaccines against other filoviruses (SUDV, BDBV, MARV) are still at pre-clinical or clinical stages. The development of vaccines requires protective efficacy studies using animal models of infection. However, animal studies using wild-type filoviruses require maximum biosafety level (BSL-4), which hinders filoviral vaccine advancement. In this study, we developed a surrogate animal model of EBOV-, SUDV-, BDBV- and MARV- GP-mediated entry using Ifnar1-knockout (-/-) mice and recombinant vesicular stomatitis viruses (rVSVs), carrying a genetic sequence of filoviral glycoprotein (GP) and pseudotyped with correspondent GP. We showed that rVSV-EBOV-GP was 100% lethal for Ifnar1(-/-) mice after inoculation by multiple routes, and its active propagation was within 24 h post-infection. We performed dose-dependent lethality assessment in small groups to estimate median lethal doses for rVSV-EBOV-GP, rVSV-SUDV-GP, rVSV-BDBV-GP, rVSV-MARV-GP in Ifnar1-knockout mice and showed high viral load in liver and immune cell reservoirs. The developed models contribute to safe and effective research of filoviral vaccines and therapeutic antibodies under BSL-2 conditions.

Adenovirus vector vaccines became an important class of vaccines following their approval for the prevention of Ebola virus fever and COVID-19. Quality dat

Background/Objectives: Flaviviruses, including West Nile virus (WNV), pose global health challenges due to their worldwide distribution, pathogenicity, and lack of effective treatments or vaccines. Today, WNV is considered the most important causative agent of viral encephalitis worldwide. This study investigated the different forms of the main WNV antigen—the preM/E protein—in the context of its immunogenic and protective properties. Methods: The recombinant adenovirus type 2 (rAd2) vectors expressing different forms of the WNV preM/E genes were obtained using standard molecular biology techniques. Immunogenicity in mice was assessed by enzyme-linked immunosorbent assay (ELISA) and virus neutralization assay. Immunological efficacy was evaluated in a mouse viral challenge model. Results: The rAd2 vector expressing the West Nile virus preM/E gene with mutations in the fusion loop exhibited robust immunogenicity when administered intramuscularly either once or in a homologous prime-boost regimen. This antigen form, as part of an adenoviral vector, protected mice from death in viral challenge experiments, providing 100% survival following WNV challenge. Conclusions: We believe that a vaccination strategy involving a recombinant adenoviral vector based on human adenovirus type 2 and the WNV antigen represented by the preM/E gene with mutations in the fusion loop may be a promising approach for combating West Nile virus infection.

Despite the widespread accessibility of vaccines and antivirals, seasonal influenza virus epidemics continue to pose a threat to public health. In this study, we constructed a recombinant replication-deficient simian adenovirus type 25 vector carrying the full-length hemagglutinin (HA) of the H1N1 influenza virus, named rSAd25-H1. Both systemic and mucosal humoral immune responses, as well as the protective efficacy, were assessed in mice immunized via the intramuscular (IM) or intranasal (IN) route. A single-dose IM or IN administration of rSAd25-H1 elicited a robust systemic IgG antibody response, including hemagglutination inhibition antibodies. As expected, only IN immunization was able to induce IgA production in serum and respiratory mucosa. Notably, a single dose of rSAd25-H1 at the highest dose (10¹⁰ viral particles) conferred complete protection against lethal homologous H1N1 challenge in mice despite the route of administration. These findings demonstrate the potential of simian adenovirus type 25-based vectors as a promising candidate for intranasal vaccine development targeting respiratory pathogens.

Introduction. In 2022, the World Health Organization declared monkeypox a public health emergency. The monkeypox virus (MPV) is part of the Orthopoxvirus genus within the Poxviridae family. During replication, orthopoxviruses produce two distinct forms of viral particles: the extracellular enveloped virion (EEV), released via exocytosis, and the intracellular mature virion (IMV), released through cell lysis. These forms differ in surface proteins composition, influencing their immunogenicity and infectivity. Aim. To evaluate the immunogenic and protective activity of nine surface antigens of vaccinia virus. Materials and methods . Recombinant human adenoviruses type 2 (rAd2) carrying surface antigens of vaccinia virus were obtained using homologous recombination in bacteria, followed by adenoviral particle assembly in HEK293 cells. The immunogenic and protective properties of these adenoviruses were tested in BALB/c mice. The presence of antibodies to the vaccinia virus was assessed using ELISA, and survival rates were evaluated in a lethal infection model after intranasal challenge with the vaccinia virus strain Western Reserve. Results. The most immunogenic and protective antigens of the vaccinia virus within rAd2 were glycoprotein B5 of the EEV and membrane-associated protein H3 of the IMV, both showing 100% protective efficacy after intranasal immunization. Conclusion. Using a panel of recombinant adenoviruses carrying genes of vaccinia virus surface proteins, it was shown that optimal protection is achieved using a combination of enveloped and mature virion antigens. This method could be used for development of new multivalent preparations against various viral infections.

Lassa hemorrhagic fever is an acute human infectious disease with high mortality rate and pandemic potential. To date, there are no approved drugs for the specific treatment or prevention of Lassa hemorrhagic fever in the world. The aim of this study was to develop and evaluate the immunobiological properties and preclinical safety of a candidate vaccine for the prevention of Lassa hemorrhagic fever (LHF) based on recombinant human adenoviral vectors. Standard genetic engineering techniques, molecular biology techniques, virological methods, and animal testing procedures were used in the course of the study. A combined vector candidate vaccine for the prevention of Lassa hemorrhagic fever has been designed and characterized. The vaccine is composed of two components for heterologous immunization in a prime-boost regimen. Both components are based on recombinant replication-defective adenovirus vectors. The first component of the vaccine is a recombinant human adenovirus type 26; the second component is a recombinant human adenovirus type 5. Both recombinant vectors contain the codon-optimized sequence of Lassa virus glycoprotein. Two experimental batches of the candidate vaccine were produced under GMP-conditions and analyzed. The results of studies in compliance with appropriate specifications for viral vector vaccines are provided. In preclinical studies in mice, antigen-specific IgG response was detected after immunization with two vaccine components, either separately, or in a prime-boost regimen. The time dynamics of the IgG response was also studied on 42, 77, 119 and 147 days after immunization. At the same time, despite achieving 100% seroconversion, no virus neutralizing antibodies were detected in any of the samples collected from immunized mice. A biodistribution study showed that 24 hours following intramuscular injection of the vaccine components, the DNA of adenovirus vectors was detected only at the site of injection and in regional lymph nodes. Based on preclinical safety assessments (including acute toxicity, chronic toxicity, immunotoxicity, allergenic properties, reproductive toxicity), no contraindications were found for initiation of the candidate Lassa vaccine clinical trials. Taken together, the results demonstrate that the candidate vaccine for prevention of Lassa hemorrhagic fever based on recombinant adenovirus vectors types 26 and 5 is a promising drug for specific immunoprophylaxis.

Background: The use of adenoviral vector vaccines is an effective way to induce an antiviral immune response. Most vaccines used worldwide are based on human adenovirus type 5 (Ad5). Pre-existing natural immunity to Ad5 is a limitation to the frequent use of Ad5-based vectors in gene therapy and vaccination. In the present study, we assessed the seroprevalence and cross-reactivity of neutralizing antibodies to six adenovirus types: Ad1, Ad2, Ad5, Ad19, Ad26, and Ad37. Methods: The study used immunological and virological research methods. Adenovirus vectors encoding the enhanced green fluorescent protein (EGFP) were developed by the N.F. Gamaleya National Research Center for Epidemiology and Microbiology. For seroprevalence and cross-neutralation study we used a panel of sera from 101 volunteers. Results: The conducted correlation analysis showed a strong relationship between seropositivity and the geometric mean titer of neutralizing antibodies to adenovirus. An increase in antibody titers indicates the formation of a humoral immune response and population immunity. We determined that there is no cross-reactivity between NtAbs to Ad1, Ad2, Ad5 and Ad26. Conclusions: Based on the presented data, we assume that Ad1 or Ad2 should be considered the most promising alternative candidates for the development of vaccine vectors.

This work highlights the importance of nucleotide sequence optimization of a target gene in the development of vaccines. The purpose of the study was to co

Introduction. Recombinant adenoviruses are widely used in the development of vaccines for a variety of infectious diseases. Despite numerous clinical studies, only a few types of human (types 5 and 26) and simian (isolate Y25) adenoviruses are currently used to produce vaccine formulations. Different types of adenoviruses vary in their cellular tropism, which plays a key role in their ability to elicit an immune response. The aim of this study was to investigate the cellular tropism of the simian adenovirus type 25 in vitro and its biodistribution in vivo in comparison with human adenoviruses types 5 and 26. Materials and methods. The efficiency of in vitro transduction was evaluated on 15 different cell lines using recombinant adenovirus vectors expressing the enhanced green fluorescent protein (EGFP) reporter gene. In vivo biodistribution and bioluminescence imaging were evaluated in BALB/c mice after administration of recombinant adenoviral vectors encoding the luciferase reporter gene. The acute toxicity of a recombinant simian adenovirus type 25 vector was assessed in mice and rats following intramuscular or intravenous administration. Results. Recombinant simian adenovirus effectively transduces a wide range of cells. At the same time, a higher tropism to human glioblastoma cells (GL-6) was found compared to the other two studied adenoviruses. In vivo experiments have shown that recombinant adenoviruses are mainly localized at the injection site, and transgene expression persists for 21 days. Acute toxicity studies demonstrated that simian adenovirus type 25 vector was well-tolerated, with no animal deaths or detectable toxic effects. Conclusion. The new platform based on the recombinant simian adenovirus type 25 is not inferior to the existing and well-established delivery systems based on human adenovirus types 5 and 26. Due to its high level of gene transfer and favorable safety profile, the use of the simian adenovirus type 25 in medicine has the potential to offer many benefits for the development of vaccines against future infectious diseases.

Background: The SARS-CoV-2 virus continuously acquires mutations, leading to the emergence of new variants. Notably, the effectiveness of global vaccination efforts has significantly declined with the rise and spread of the B.1.1.529 (Omicron) variant. Methods: The study used virological, immunological and histological research methods, as well as methods of working with laboratory animals. In this study, we evaluated the Gam-COVID-Vac (Sputnik V), an adenoviral vaccine developed by the N.F. Gamaleya National Research Center for Epidemiology and Microbiology, and conducted experiments on hemizygous K18-ACE2-transgenic F1 mice. The variants studied included B.1.1.1, B.1.1.7, B.1.351, B.1.1.28/P.1, B.1.617.2, and B.1.1.529 BA.5. Results: Our findings demonstrate that the Sputnik V vaccine elicits a robust humoral and cellular immune response, effectively protecting vaccinated animals from challenges posed by various SARS-CoV-2 variants. However, we observed a notable reduction in vaccine efficacy against the B.1.1.529 (Omicron BA.5) variant. Conclusions: Our results indicate that ongoing monitoring of emerging mutations is crucial to assess vaccine efficacy against new SARS-CoV-2 variants to identify those with pandemic potential. If protective efficacy declines, it will be imperative to develop new vaccines tailored to current variants of the virus.