Ebola hemorrhagic fever: Properties of the pathogen and development of vaccines and chemotherapeutic agents

Molecular Biology - Ebola hemorrhagic fever (EHF) epidemic currently ongoing in West Africa is not the first among numerous epidemics in the continent. Yet it seems to be the worst EHF epidemic...     

Impact of mutations in nucleoprotein on replication of influenza virus A/Hong Kong/1/68/162/35 reassortants at different temperatures

The nucleoprotein (NP) of influenza virus is a multifunctional RNA binding protein. The role of NP in the adaptation of influenza viruses to a host has been experimentally proved. Ambiguous data are available on the role of nucleoprotein in the attenuation of influenza A viruses, which is characterized by ability to replicate at low temperature (26°C) and inability to replicate at high temperature (39°C). Influenza virus donor strain A/Hong Kong/1/68/162/35 (H3N2), adapted to growth at low temperature, differs from the wild type virus by 14 amino acid mutations in the internal and non-structural proteins. Two mutations occurred in the NP: Gly102Arg and Glu292Gly. We have obtained viruses with point reverse-mutations in these positions and compared their replication at different temperatures by measuring infectious activity in chicken embryos. It has been shown that reverse mutation Gly292Glu in the NP reduced virus ability to replicate at low temperature, the introduction of the second reverse mutation Arg102Gly completely abolished virus cold adaptation.     

Expression in Plants of a Recombinant Protein Based on Flagellin Linked to Conservative Fragments of M2 Protein and Hemagglutintin of Influenza Virus

The composition of traditional influenza vaccines require nearly annual updates due to the high variability of the influenza virus. The use of conservative viral antigens, the extracellular domain of the transmembrane protein M2, and fragments of the second subunit of hemagglutinin provides the opportunity to create recombinant broad-spectrum vaccines. Bacterial flagellin was used as a mucosal adjuvant to increase the immunogenicity of these conservative antigens. Recombinant proteins based on flagellin simultaneously containing M2e and a fragment of the hemagglutinin stem region were expressed in Nicotiana benthamiana plants using a self-replicating vector based on the potato virus X genome. Methods for their isolation from plants and purification have been developed. The developed expression system can be used to produce a new candidate influenza vaccine in plants.     

Development of the anti-influenza H5N1 vaccines worldwide and in Russia

Article is dedicated to analytical investigation of the problem of current technologies in construction and manufacturing of anti-influenza vaccines. Epidemiological events in July-November 2005 in Russia (mainly in Siberia) and later in Ukraine showed that Health Care system was not ready for that turn over of epidemiological situation. It was completely the same situation in other countries. There are two general questions of a readiness in pre-pandemic situation: level of a diagnostic monitoring of epidemiological situation and preparedness to fast production of actual vaccine preparations. First task can be solved by immediate production of diagnostic sets for regional branches of National WHO Centers, and a second one depends on application of a novel approaches in construction of a anti-influenza vaccines. The construction of anti-influenza vaccines is based on genetic engineering (reverse genetics) and manipulation with plasmids carried out basic viral genes. Reassortation technology for preparation of hybrid viruses is going to the past by objective reasons. Advanced technologies are safety in laboratories and in manufacturing facilities. Moreover, genetic engineering in this field allows to planing the construction of vaccines bank, when the prognoses for actual viruses include more then two strains with different antigenic properties.     

HLA system antigens in persons with differing susceptibility to the causative agents of acute respiratory diseases

The relationship between the susceptibility of the body to infections caused by influenza A and B viruses, parainfluenza viruses, adenoviruses, Mycoplasma pneumoniae and antigens of the HLA system was studied on a group of 400 adolescents placed under clinico-epidemiological surveillance for two years. The relationship between histocompatibility antigens and acute respiratory diseases was manifested in a decrease or increase in the occurrence of recurrent diseases and infections or in the probability of the development of the diseases in infected persons. HLA B40 was associated with resistance to influenza A, B18 and B21 were associated with resistance to parainfluenza, B15 and B35 were associated with resistance to M. pneumoniae infection; susceptibility to influenza B was registered in persons with HLA B12 and to M. pneumoniae infection, in persons with HLA B16 and B18. With respect to different infective agents, the relative risk of infection varied within 1.7 and 5.0.     

Plant-produced recombinant influenza vaccine based on virus-like HBc particles carrying an extracellular domain of M2 protein

Conventional influenza vaccines are based on a virus obtained in chicken embryos or its components. The high variability of the surface proteins of influenza virus, hemagglutinin and neuraminidase, requires strain-specific vaccines matching the antigenic specificity of newly emerging virus strains to be developed. A recombinant vaccine based on a highly conservative influenza virus protein M2 fused to a nanosized carrier particle can be an attractive alternative to traditional vaccines. We have constructed a recombinant viral vector based on potato X virus that provides for expression in the Nicotiana benthamiana plants of a hybrid protein M2eHBc consisting of an extracellular domain of influenza virus M2 protein (M2e) fused to hepatitis B core antigen (HBc). This vector was introduced into plant cells by infiltrating leaves with agrobacteria carrying the viral vector. The hybrid protein M2eHBc was synthesized in the infected N. benthamiana plants in an amount reaching 1–2% of the total soluble protein and formed virus-like particles with the M2e peptide presented on the surface. Methods of isolation and purification of M2eHBc particles from plant producers were elaborated. Experiments on mice have shown a high immunogenicity of the plant-produced M2eHBc particles and their protective effect against lethal influenza challenge. The developed transient expression system can be used for production of M2e-based candidate influenza vaccine in plants.     

Importance of secretory antibodies in nonsusceptibility to influenza B disease and the prevention of the spread of this virus

Under the conditions of the natural outbreak of influenza B a rise in the level of antibodies in the blood serum and respiratory tract secretions has been found to occur in nonimmunized persons during this disease. Secretory antibodies, along with serum antibodies and in interaction with them, prevent the development of clinically manifest influenza. The asymptomatic form of influenza infection has been revealed: during this form of the disease the response of the body is limited to the local immunity system.     

Role of genetic factors in the epidemic process in respiratory viral infections

Observations carried out during 1973-1979 indicate that persons, who are frequently ill, determine the incidence of influenza and acute respiratory diseases among various groups of adult population at all epidemic periods. In constantly observed groups of different ages such persons were the source of 60-85% of the outbreaks of acute respiratory diseases. The correlation between genotypic blood markers (the ABO and HLA systems) and susceptibility to respiratory viruses has been established.     

Antigenic anachronism of the influenza A(H2N2) viruses in Leningrad in 1980. I. The epidemiological and serological characteristics of the influenzal infection caused by A/Leningrad/80 (H2N2) viruses

In April-May 1980 a number of unrelated outbreaks of influenza-like diseases were registered in Leningrad in an infant home (50 out of 68 children under observation, aged 3 months to 2 years, were affected) and among the pupils of a boarding school (13 out of 50 adolescents under observation, aged 15-17 years, were affected). 5 strains of influenza A virus were isolated from 3 sick children and 1 clinically healthy child. A similar virus was isolated from a sick adolescent in a boarding school, as well as from a female patient aged 24 years at a domiciliary focus of infection (a sporadic case). In the subsequent laboratory investigation all these 7 strains were identified as viruses A/H2N2. Isolated cases of seroconversion to hemagglutinin H2 were definitely registered in 6 patients during February--May 1980. In 3 cases, including the 24-year old female patient with an acute respiratory disease, seroconversion to hemagglutinin H2 was observed in combination with the release of influenza viruses A/H2N2 from these patients. 2 influenza virus strains with this antigenic characteristic were isolated from a young female patient at an interval of 3 days. Among the patients admitted to the clinics of the Research Institute of Influenza in Leningrad on account of acute respiratory diseases isolated cases of diagnostically significant seroconversion to hemagglutinin H2 constituted 3.5% among children and 4.5% among adults. The study of the level of antihemagglutinins in the population revealed that in 1980 persons aged 18-50 years showed a high level of antihemagglutinins.(ABSTRACT TRUNCATED AT 250 WORDS)     

Molecular-biological characteristics of rubella virus strains isolated in St. Petersburg

In the surveillance of the incidence of rubella in the northwestern region of Russia, an analysis was conducted of swabs from the noses and pharynxes of 37 patients with rubella treated in Solovev District Military Clinical Hospital No. 442. To isolate the virus, the RK-13 cell line was used, in total 22 strains were allocated. The strains of the Rubella virus isolated in St. Petersburg during the 2007 outbreak belong to the 1E genotype. In the culture of RK-13 cells, changes of cellular morphogenesis and the formation of replicative complexes, as well as the shells of virions of rubella viruses, were detected.