ABSTRACT
BACKGROUND: Recombinant hemagglutinin (rHA) and neurominidase (rNA) developed in our investigation are amino acid sequence consensus variants of H1N1 2009 subtype influenza virus strain, also including immunogenic epitopes typical for other influenza virus subtypes (H3N1 and H5N1). Substitutions were made: typical for Russian virus isolates (in HA - S220T, NA - D248N) and in active centers of molecules - R118L, R293L, R368L; C92S, C417S to increase recombinant proteins stability in E. coli. The aim of the present work was to study immunogenicity of the obtained rHA and rNA. MATERIAL AND METHODS: Fragments aa 83-469 of NA and aa 61-287 of HA were chosen because they include the main B-cell epitopes and are the minimal structures for correct folding of target proteins. The designed nucleotide sequences were synthesized and purified and the expression of rNA and rNA were analyzed. For immunization and virus challenge we used influenza viruses A/California/04/2009 (H1N1), A/PR/8/34 (H1N1), A/Perth/16/2009 (H3N2), A/Chicken/Kurgan/05/2005 R.G. (H5N1), and B/Florida/04/2006. Specific IgG levels were determined by ELISA in 96-well ELISA plates. Significant differences of survival in mouse groups were analyzed by Mantel-Cox (log-rank) and Gehan-Breslow-Wilcoxon tests. RESULTS: The obtained results demonstrate the high immunogenicity and ability of indicated proteins mixture to provide similar cross-protection against influenza viruses of the H1N1 subtype. CONCLUSIONS: The data obtained suggest efficient pluripotent vaccine creation based on HA and NA conservative regions.
Subject(s)
Epitopes/immunology , Hemagglutinin Glycoproteins, Influenza Virus/immunology , Influenza A virus/immunology , Neuraminidase/immunology , Recombinant Proteins/immunology , Animals , Body Weight , Hemagglutinin Glycoproteins, Influenza Virus/isolation & purification , Immunization , Immunoglobulin G/blood , Mice , Neuraminidase/isolation & purification , Orthomyxoviridae Infections/blood , Orthomyxoviridae Infections/immunology , Orthomyxoviridae Infections/prevention & control , Orthomyxoviridae Infections/virology , Plasmids/metabolism , Recombinant Proteins/isolation & purification , Survival AnalysisABSTRACT
Human immunodeficiency virus (HIV)-1 subtype A strains circulating among the majority of HIVinfected individuals in the former Soviet Union (FSU) countries demonstrate low genetic diversity. The consensus sequence of the FSU region-specific isolate has been used for the candidate DNA vaccine development. We constructed recombinant plasmids with four viral genes: env (gp140), gag, pol (reverse transcriptase), and nef. We immunized BALB/c mice intramuscularly using equimolar mixture of four recombinant plasmids, and observed significant cytotoxic T lymphocyte response and specific CD8(+) cell production against cells presenting HIV-1 peptides. Overall, the Th1 pathway of immune response clearly dominated. Immunological properties of this candidate DNA vaccine against HIV-1 suggest the possibility of its further study in clinical trials.
Subject(s)
HIV Infections/immunology , HIV Infections/prevention & control , HIV-1/genetics , HIV-1/immunology , Vaccines, DNA/immunology , Vaccines, DNA/therapeutic use , Animals , Base Sequence , Drug Design , HIV Infections/epidemiology , HIV Infections/genetics , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Prevalence , Russia/epidemiologyABSTRACT
BACKGROUND: Earlier we suggested the concept of the positive evolutionary role of tumors. According to this concept, tumors provide conditions for the expression of evolutionarily new and/or sleeping genes in their cells. Thus, tumors are considered as evolutionary proving ground or reservoir of expression. To support this concept we have previously characterized in silico and experimentally a new class of human tumor-related transcribed sequences. RESULTS: In this article we describe results of further studies of previously described tumor-related sequences. The results of molecular phylogeny studies, Southern hybridization experiments and computational comparison with genomes of other species are presented. CONCLUSION: These results suggest that these previously described tumor-related human transcripts are also relatively evolutionarily new.
