[The Challenges of Vaccine Development Against Betacoronaviruses: Antibody Dependent Enhancement and Sendai Virus as a Possible Vaccine Vector].

To design an effective and safe vaccine against betacoronaviruses, it is necessary to elicit a combination of strong humoral and cell-mediated immune responses as well as to minimize the risk of antibody-dependent enhancement of viral infection. This phenomenon was observed in animal trials of experimental vaccines against SARS-CoV-1 and MERS-CoV that were developed based on inactivated coronavirus or vector constructs expressing the spike protein (S) of the virion. The substitution and glycosylation of certain amino acids in the antigenic determinants of the S-protein, as well as its conformational changes, can lead to the same effect in a new experimental vaccine against SARS-CoV-2. This review outlines approaches for developing vaccines against the new SARS-CoV-2 coronavirus that are based on non-pathogenic viral vectors. For efficient prevention of infections caused by respiratory pathogens the ability of the vaccine to stimulate mucosal immunity in the respiratory tract is important. Such a vaccine can be developed using non-pathogenic Sendai virus vector, since it can be administered intranasally and induce a mucosal immune response that strengthens the antiviral barrier in the respiratory tract and provides reliable protection against infection. The mucosal immunity and the production of IgA antibodies accompanying its development reduces the likelihood of developing an antibody-dependent infection enhancement, which is usually associated only with immunopathological IgG antibodies.

[Apoptin enhances the oncolytic activity of vaccinia virus].

Chicken anemia virus gene encoding apoptin, a selective killer of cancer cells was synthesized and inserted into vaccinia virus (strain L-IVP) genome. The insertion has replaced major part of the viral C11R gene encoding viral growth factor (VGF), which is important for the virulence. The recombinant virus VVdGF-ApoS24/2 was obtained through the transient dominant selection technique with the use of puromycin resistance gene as the selective marker. The expression apoptin gene from a synthetic early-late promoter of vaccinia virus effectively provides accumulation of the protein in the cells infected with the VVdGF-ApoS24/2 virus. Despite the presence of virus growth factor signal peptide at apoptin N-terminal secretion of the recombinant protein into culture medium did not occur. The recombinant virus VVdGF-ApoS24/2 was found to have a significantly greater selective lyticactivity on human cancer cell lines (A549, A431, U87MG, RD and MCF7) as compared with the parent strain L-IVP and its variant VVdGF2/6 with the deletion of the C11R gene. The results suggest that the use of apoptin represents a promising approach for improving the natural anticancer activities of vaccinia virus.

[The hepatitis A virus: structural and functional organization of the genome, its molecular diagnostic value and cultivation].

The analysis of recently published data on hepatitis A virus (HAV) genome clinical features, molecular diagnostic value and cell culture propagation are reviewed. The growing need in the study of the genetic diversity of HAV isolates and the search of its possible new antigenic variants are underlined. The results of the cultivation of different HAV strains are analyzed for possible application in vaccine and diagnostic kit production.

[Oncolytic properties of some orthopoxviruses, adenoviruses and parvoviruses in human glioma cells].

UNLABELLED: Currently one of the most promising approaches in development of cancer virotherapy is based on the ability of oncolytic viruses to selective infection and lysis of tumor cells. AIM: The goal of the study was to identify and evaluate perspective oncolytic viruses capable of selectively destroying human glioma cells. PATIENTS AND METHODS: Original GB2m, GA14m and GB22m glioma cell cultures derived from patients were used for evaluating in vitro oncolytic activity of some typical orthopoxviruses, adenoviruses and parvoviruses. RESULTS: The oncolytic activity in the human glioma cell models was confirmed for LIVP and WR strains of vaccinia virus, Adel2 and Ad2del strains with deletions within E1B/55K gene and derived from human adenoviruses type 2 and 5, respectively. CONCLUSIONS: We consider these oncolytic viruses as promising agents for the treatment of human malignant glioma.

[Oncolytic adenoviruses in anti-cancer therapy: current status and perspectives].

Lytic viral infection results in production of viral progeny, and lysis of the infected cells. Tumor cells are usually more sensitive to virus infection. Studies of viral oncolysis indicate that it could represent a promising alternative approach to cancer therapy. The ability of viruses to kill selectively cancer cells had been noticed for quite a long time ago. However, only in recent years, based on deeper understanding of molecular biology of viruses and the cell and due to the development of modern methods for directed modification of viruses, there emerged a real opportunity for development of virus variants with improved therapeutic potential. Adenoviruses represent one of the most studied models of oncolytic viruses. The DNA-containing viruses are very suitable for genetic manipulation and show minimal pathogenicity. The review summarizes data on directions and approaches aiming generation of highly efficient variants of oncolytic adenoviruses. The approaches include introduction of directed genetic modifications into viral genome, accelerated selection of oncolytic viral variants following treatment with mutagens, the use of adenoviruses as vectors for introduction of therapeutic gene products, optimization of viral delivery systems, minimalization of negative effects from the host immune system etc. The dynamic development of studies in the field holds promise for introduction into clinical practice of many variants of oncolytic adenoviruses in the very near future.

[Oncolytic viruses in the therapy of gliomas].

Despite the advances of modern medicine, malignant glioblastoma cure remains an elusive goal. Both the invasive nature and location in vital areas of the brain make this type of tumors difficult for surgical treatment, while the current adjuvant therapy is not as successful as expected. Frequent recurrence and invasiveness of malignant gliomas is due to resistance of glioma stem cells to conventional radiation and chemotherapy. Technological advances in constructing recombinant viruses have allowed creating strains with high oncolytic activity toward glial tumors. Many of these strains have passed Phase I of clinical trials and demonstrated high safety. Despite the obvious potential of the approach, efficiency of the existing strains is still far from being sufficient for effectively curing the disease and require further improvement. The review summarizes results obtained with the most successful variants of oncolytic viruses that come down to the clinical trials and discusses the prospects for new approaches in virotherapy of malignant gliomas.

[Oncolytic enteroviruses].

Increasing information concerning molecular biology of viruses and virus-cell interactions makes it possible to use viruses as a tool in effort to treat cancer diseases. As a rule, tumor cells are highly sensitive to viruses that may be used in cancer therapy. Therewith, applications of viral oncolysis in treatment of cancer diseases assume maximum possible safety of used viruses for patient and environment. Human enteroviruses are one of the most convenient sources to generate oncolytic viruses. Many of enteroviruses are non-pathogenic for humans or cause mild disease. Progress in genetic engineering permits to develop attenuated enterovirus variants with high safety and selectivity. This review focuses on the main members of Enterovirus genus, such as Coxsackieviruses, and vaccine strains as promising source for development of oncolytic agents, applicable for cancer therapy. It reviews data concerning recently developed and tested oncolytic variants of enteroviruses and discusses perspectives of their application in cancer therapy and problems, concerning their improvement and practical use.

[Oncolytic poxviruses].

The latest data on selection and construction of poxviruses capable of specifically lysing tumor cells of different genesis, inducing antitumor immunity and apoptosis of malignant cells are discussed. The review concerns several directions: virus attenuation, insertion of immunomodulatory protein genes, and anti-tumor protein genes. Thymidine kinase and viral growth factor genes make the greatest contribution to the virus attenuation as their inactivation results in the virus inability to replicate in non-dividing cells, thereby contributing to increased selectivity with respect to tumor cells. Among the immunomodulatory proteins, interleukins 2, 12, and granulocyte-macrophage colony-stimulating factor proved to be most promising for oncolytic virotherapy. An attempt to use p53 protein gene expressed by vaccinia virus for addressed apoptosis of tumor cells was reported. The use of the double and triple viral recombinants carrying genes of multidirectional action seems to be most promising. Encouraging results were obtained using vaccinia virus in the oncotherapy with prodrugs and angiogenesis inhibitors. At present, two poxviral strains are undergoing Phase III clinical trials as anti-tumor preparations in the USA.

[Genetic diversity of group A rotavirus isolates found in Western Siberia in 2007-2011].

Genetic analysis of group A rotavirus recovered from fecal samples of children admitted to hospitals in Novosibirsk and Omsk during four epidemic seasons 2007, 2007/2008, 2009/2010, 2010/2011 was performed. A total of 1416 rotavirus isolates were genotyped using multiplex PCR. The isolates of the most common rotavirus genotypes G1P[8], G4P[8], G2P[4], G3P[8] co-circulated in Western Siberia during 2007-2011. In isolated cases G9P[8], G2P[8], G3P[9], and G4P[6] genotypes were detected. Change of dominant genotype from G1P[8] to G4P[8] occurred in 2008 in Omsk and in Novosibirsk in 2009 as well. Incidence and distribution of rotavirus genotypes differed and changed every epidemic season in both cities. The phylogenetic analysis based on VP4 (VP8*), VP7, and VP6 gene sequences showed that the majority of isolates from Novosibirsk and Omsk were clustered together and demonstrated high level homology with rotavirus isolates found in other regions of Eurasia. In addition, a rare P[8]b (OP354-like) subtype of the VP4 gene was identified in fourteen isolates (G9, G1, and G4) in Novosibirsk and in a single isolate Omsk08-381/G9P[8]b in Omsk. The results obtained in this study demonstrate the necessity of long-term monitoring of rotavirus isolates in Western Siberia. This is important for selection of rotavirus vaccine for immunization of infants, improvement of diagnostic kits and understanding of the epidemiology and the evolution of group A rotaviruses.

[Oncolytic parvoviruses. A new approaches for cancer therapy].

Parvoviruses such as parvovirus H-1 (H-1PV) may selectively infect and lysis cancer cells. The parvoviruses also induce an immune system to eliminate the tumor cells through the formation of anti-cancer immunity. One of the possible mechanisms of antitumor activity is associated with the direct induction of apoptosis by parvoviral proteins NS1 and 11 kDa. Parvovirus-based vectors are promising for gene therapy of oncological diseases and genetic disorders in humans. Parvoviruses were successfully used for the experimental treatment on animal models of human glioma, neuroblastomas, lymphomas, pancreatic carcinoma, carcinomas and breast tumors. ParvOryx is the first oncolytic preparation constructed on the base of H-1PV; its phase I/IIa clinical trials in patients with glioblastoma multiforme are in process.

[Outbreak of acute enterovirus intestinal infection in Sakhalin region in August 2010].

The investigation of cases of acute intestinal infections in the Sakhalin region of Russia in August, 2010 is described. Epidemiological and molecular biological studies were conducted. After initial PCR screening and determining the nucleotide sequences of the positive samples the following enteroviruses were found: Coxsackie A2 - 42 samples (45%), Coxsackie A4--31 sample (34%), Enterovirus 71--6 samples (6,5%), Coxsackievirus B5--6 samples (6,5%), Coxsackie B3--4 samples (4%) and Coxsackie B1--4 samples (4%). The phylogenetic analysis of sequences showed that the closest analogues for the nucleotide sequences of these genotypes were previously identified in Japan, Korea and China in 2000-2010.

COVID-19, Influenza, and Other Acute Respiratory Viral Infections: Etiology, Immunopathogenesis, Diagnosis, and Treatment. Part I. COVID-19 and Influenza.

The paper briefly reviews pathogens causing acute respiratory viral infections (ARVIs), including influenza viruses; coronaviruses, including SARS-CoV-2; parainfluenza viruses, adenoviruses, pneumoviruses, and specifically respiratory syncytial virus and metapneumoviruses, enteroviruses, rhinoviruses, and bocaviruses. This review presents modern data on the structure and replication of viruses, epidemiology, and immunopathogenesis of diseases and on diagnostics, preventive vaccination, and antiviral drugs for the treatment of ARVIs. Special attention is paid to the SARS-CoV-2 virus caused COVID-19 pandemic with analyses of similarities and differences between COVID-19 and other ARVIs, first of all, influenza virus. Topical issues regarding ARVI vaccination and the search for new broad-spectrum antiviral drugs are discussed.

COVID-19, Influenza, and Other Acute Respiratory Viral Infections: Etiology, Immunopathogenesis, Diagnosis, and Treatment. Part 2. Other Acute Respiratory Viral Infections.

The first part of this paper presented the current knowledge on two very significant respiratory diseases with high pandemic potential, COVID-19 and influenza. The second part reviews other pathogens that cause acute respiratory viral infections, ARVI, including parainfluenza viruses, adenoviruses, pneumoviruses and specifically respiratory syncytial virus, enteroviruses, rhinoviruses, bocaviruses, and seasonal coronaviruses. The review presents modern data on the structure and replication of viruses, epidemiology and immunopathogenesis of diseases, diagnostics, preventive vaccination, and antiviral drugs. Topical issues regarding ARVI vaccination and the search for new broad-spectrum antiviral drugs are discussed.

[The production of high-yielding strain of rubella virus from wild type virus extracted from a patient in the Western Siberia in 2006].

The study was targeted to investigate the propagation of rubella virus in the cell cultures of various origins and with different cultivation methods. The high-yielding strain of rubella virus was produced. The "spinner-culture" cultivation method was applied and the strain's RNA was detected in 10-8 dilution in real time mode. This strain is supposed to be used in preparation of the standard antigen to implement in the development of immune enzyme test system targeted to the rubella virus specific antibodies.

[Complete genomic sequence of rapidly replicating strain MB-7 of hepatitis a virus and its characterization in comparison with nucleotide sequences of other hepatitis A virus strains].

Complete nucleotide sequence of genomic RNA of hepatitis A virus (HAV) rapidly replicating strain MB-7 was determined. Comparison of nucleotide and deduced amino-acid sequences demonstrated the highest level of identity of MB-7 with strain HAS-15 (above 99%) and high homology with other HAV strains (HM 175/7, CR326, and GBM/HFS) used in production of anti-hepatitis A vaccines. MB-7 was classified as subgenotype IA. Phylogenetic analysis showed that MB-7 is most closely related to the strain HAS-15 and the HAV variants circulating in Russia. Comparative analysis of genomic differences between MB-7 and HAS-15 with other HAV strains revealed among changes characteristic of MB-7 those typical of the described earlier rapidly replicating HAV strains (nt. 149-162 in 5'-untranslated region and changes in the VP3 and 2C genes). These results suggest the functional importance of changes in above-mentioned regions of HAV genome for the increased replication level of MB-7 in vitro.

[Molecular genetic characteristics of astroviruses circulating in Novosibirsk].

A total of 1107 fecal samples from young children admitted to hospital for acute enteric infection in January to December 2007 were tested for astroviruses. Astroviruses were detected in 64 (5.8%) of the 1107 stool samples, only 50% of them were found as monoinfections. Astroviruses were recorded throughout the year; however, no seasonality for this infection could be ascertained. Cases of astrovirus infection were mainly observed in infants under one year of age (90%). Astroviruses were typed sequencing the ORF2 fragment; only HAstV-1 and HAstV-2 were found in Novosibirsk.

The New Coronavirus COVID-19 Infection.

In December 2019, the first cases of pneumonia of unknown etiology were found in Wuhan (China). Later, the pneumonia was associated with a new coronavirus; in February 2020, the World Health Organization (WHO) gave the name COVID-19 to the new disease, while the International Committee on Taxonomy of Viruses (ICTV) gave the name SARS-CoV-2 to the virus causing it. By March 11, 2020, when the virus had spread to 114 countries, the number of diagnosed patients had reached 118 thousand and the number of deaths was 4000, the WHO declared the outbreak of the disease a pandemic. In this review, we summarize the relevant information about the origin and spread of SARS-CoV-2, its epidemiology and diagnostics, and the clinical course and treatment of COVID-19.

The Challenges of Vaccine Development against Betacoronaviruses: Antibody Dependent Enhancement and Sendai Virus as a Possible Vaccine Vector.

To design an effective and safe vaccine against betacoronaviruses, it is necessary to use their evolutionarily conservative antigenic determinants that will elicit the combination of strong humoral and cell-mediated immune responses. Targeting such determinants minimizes the risk of antibody-dependent enhancement of viral infection. This phenomenon was observed in animal trials of experimental vaccines against SARS-CoV-1 and MERS-CoV that were developed based on inactivated coronavirus or vector constructs expressing the spike protein (S) of the virion. The substitution and glycosylation of certain amino acids in the antigenic determinants of the S-protein, as well as its conformational changes, can lead to the same effect in a new experimental vaccine against SARS-CoV-2. Using more conservative structural and accessory viral proteins for the vaccine antigenic determinants will help to avoid this problem. This review outlines approaches for developing vaccines against the new SARS-CoV-2 coronavirus that are based on non-pathogenic viral vectors. For efficient prevention of infections caused by respiratory pathogens the ability of the vaccine to stimulate mucosal immunity in the respiratory tract is important. Such a vaccine can be developed using non-pathogenic Sendai virus vector, since it can be administered intranasally and induce a mucosal immune response that strengthens the antiviral barrier in the respiratory tract and provides reliable protection against infection.

[Isolation and molecular characterization of the influenza virus A/H5N1 strains isolated during outbreak of avian influenza among birds in the European part of Russia in 2005: strain with ozeltamivir-resistance mutation was found].

Isolation and characterization of the influenza virus A/H5N1 strains, isolated from chicken in the Yandovka village (Tula Region) and from wild swan near the orifice of the Volga River that died during an outbreak of avian flu in autumn 2005, were carried out. Genetic and phylogenetic analyses were performed. The goals of the analysis were to determine possible geographical origin of the strain, genetic similarity of isolated strains to earlier sequenced isolates, epidemic potential, existence of pathogenicity markers, and resistance to antiviral drugs. It was shown that the isolated influenza virus belonged to highly pathogenic variants of China origin by a reassortment of viruses genotypes Z and V circulated in poultry and wild birds. A number of molecular markers of pathogenicity to gallinaceous birds and mammals were found out. Mutations in the hemagglutinin gene promoting potentially high rate of replication in humans as well as mutations causing the resistance to amantadine/rimantadine were not found. The strain isolated from wild swan had the mutation causing resistance to tamiflu/ozeltamivir.

Pathogenesis of infectious disease of mice caused by H5N1 avian influenza virus.

The pathogenesis of a disease caused by Qinghai-like H5N1 influenza virus in BALB/c mice was studied. Clinical, morphological, and immunological characteristics of the experimental infection caused by highly pathogenic A/duck/Tuva/01/06/ (H5N1) virus are described.