[IMMUNOCHEMICAL ANALYSIS OF RECOMBINANT CHIMERIC POLYPEPTIDE OspC(gar+afz) OF BORRELIA GARINIIANIP B. AFZELI ISOLATES].

AIM: Comparative study of antigenic properties of recombinant proteins OsPCgar and OsPCafz and recombinant chimeric polypeptide OspCgar+afrz, that contains amino acid sequences of mature immune dominant OspC proteins of West-Siberian isolates of Borrelia garinii (OspCgar) and B. afzelii (OspCafz), and evaluation of possibility of their use as antigens during creation of test-systems for serodiagnostics of Lyme borreliosis (LB) on the territory of Western Siberia. MATERIALS AND METHODS: Recombinant chimeric polypeptide OSpCgar+af, and recombinant mature proteins OSPCgar and OspCafz, obtained by expression of the corresponding genes in Escherichia coli cells; purified by affinity chromatography in Ni-NTA-sepharose CL-6B and studied by EIA method for the ability to bind antibodies from sera of LB patients. RESULTS: A difference in sensitiv- ity of determination by EIA method of specific IgM and IgG against borreliae in blood sera of LB patients with localized stage of the disease during use of OspCgar,'OSPCafz and OspCgar+afz chimera as antigens was shown. Chimeric antigen OSPCgar+afz was established to show higher antigenic activity compared with each of the OspCgar or OSPCafz antigens separately. CONCLUSION: The results of the study allow to examine.the recombinant chimeric polypeptide OspCgar+afz as a pos- sible component during creation of test-systems for serodiagnostics of LB on the territory of West, Siberia.

[Study of a role of artificial mycobacterial particles in the realization of immunological processes in experimental animal tuberculosis].

The protective properties of artificial mycobacterial particles versus BCG vaccine were studied in laboratory animals with experimental tuberculosis. The findings of the decreased rate of a tuberculous process and on the increased mean life span in animals inoculated with M. bovis suggest that immunization of guinea-pigs with mycobacterial particles promotes the enhanced development of antituberculous immunity in the animals. The paper proposes a promising method for designing artificial immunogens, the high-polymer antigenic structures that imitates mycobacterial particles.

[Evaluation of reasons of the MDR M. tuberculosis strains dissemination by analysis of the rifampicin and/or isoniazid resistant isolates].

During the last years in Novosibirsk region of Russia the rate of TB patients infected by MDR strains of M. tuberculosis has been constantly increasing. This increase may occur as a result of the spontaneously mutated mycobacterium selection during treatment of patients or as a result of primary infection by the resistant M. tuberculosis, or also, as a result of both reasons in combination. If the main reason of MDR strain dissemination is selection of resistant bacterium during patient treatment, the equal apportionment of the dominated mutation into the mycobacterium genotypes would be observed. If the main reason is the primary infection by resistant M. tuberculosis, the unequal apportionment would be revealed. For deeper understanding of the main reasons of the fast MDR strains spreading in the region, the distribution of the main mutations over genotypes of strains in Novosibirsk (170 isolates) and Tomsk prison (51 isolates) was investigated. Mutations in rpoB gene associated with the rifampicin resistance and in katG (isoniazid resistance) were detected by biochips. M. tuberculosis genotypings were carried out by IS6110 PCR typing or MIRU typing, in the last method the twelve loci (MIRU 2, 4, 10, 16, 20, 23, 24, 26, 27, 31, 39, 40) have been used. The most frequent mutation in the rpoB gene was Ser531-->Leu (60-70% of the rifampicin resistant strains) and Ser315-->Thr in gene katG (80% of the isoniazid resistant M. tuberculosis). Both in Novosibirsk and in Tomsk prison the rates of clustered cases transmissions were high (69 and 63% respectively). Analysis of the distribution of the dominated mutations Ser531-->Leu (rpoB) and Ser315-->Thr (katG) revealed that all of them were detected in each clusters, but in Novosibirsk there were only two clusters, in which the percentage of strains, containing mutation Ser531-->Leu (rpoB) were higher (85.7% and 77.7% respectively, P < 0.05), then in others. Among the Tomsk prison's clusters it was revealed one in which the proportion of the Ser3 15-->Thr mutation in katGwas higher (96.4%, P < 0.05). The nonuniform distribution of the dominated mutations highlighted that the epidemic spread of drug-resistant strains of M. tuberculosis in region resulted from the selection of them during patient treatment and the subsequent transmission by TB patients.

[Study of antituberculous activity of moxifloxacin conjugate with macrophage scavenger-receptor ligand].

Mycobacterium tuberculosis is an intracellular pathogen that persists in macrophages of the human host. An approach to improving the treatment of tuberculosis is target delivery of antibiotics to macrophages using ligands to macrophage receptors. The antituberculous activity of the conjugate of the antituberculous antibiotic moxifloxacin with carboxymethylglucan was studied in vitro using the J774 macrophage cell line and peritoneal macrophages. The antituberculous activity of the conjugate was higher than of the free moxifloxacin. The target antibiotic delivery to macrophage cells in tuberculosis infection was shown perspective.

[Molecular and genetic characterization of rifampicin- and/or izoniazid-resistent strains of M. tuberculosis, isolated in Novosibirsk and Tomsk regions].

The purpose of the work was to study rifampicin- and izoniazid-resistent strains of M. tuberculosis, circulating in Western Siberia, by VNTR and IS6110 typing. The authors also studied genetic causes of resistance to these antibiotics and undertook a search of new VNTR loci, displaying polymorphism in genomes of closely related clonally-disseminated variants of Mycobacterium tuberculosis in W-Beijing family model analysis.

[Genetic polymorphism of clinical Mycobacterium tuberculosis strains circulating in the territory of Novosibirsk Region].

Mycobacterium tuberculosis strains isolated from patients treated at TB dispensary branches in different districts of Novosibirsk were studied by genetic analysis. The below molecular methods were used: 1. PCR with random primers; 2. A method based on variable number of tandem repeats in loci; 3. IS6110 inverse PCR. Thirty-five samples of genome DNA of M. tuberculosis isolated were analyzed. Each of the 3 methods detected the main group of isolates, which comprised 61.8% of closely related strains revealed by method 1, 75.8%--by method 2, and 74.3%--by method 3. The remaining clusters were represented by 1 to 4 strains. The data obtained denote a relative homogeneity of M. tuberculosis strains circulating in Novosibirsk Region. No interplay was detected between the clustering of isolates and the presence or absence of mutation in genes conditioning the resistance to antibiotics.

[Construction of peptide mimetics of an epitope of the human immunodeficiency virus (HIV-1) gp41 protein, recognized by virus-neutralizing antibodies 2F5]. / Polucheni peptidov-imitatorov épitopa belka gp41 virusa immunodefitsita cheloveka (VICh-1) uznavaemogo virusneitralizuiushchimi antitelami 2F5.

A phase peptide library was screened with virus-neutralizing monoclonal antibodies (MCA) 2F5 which recognize a conserved epitope of HIV-1 gp41. Phages that expose peptides specifically binding with MCA 2F5 were selected by ELISA. Amino acid sequence analysis revealed a homology to region 662-671 of HIV-1 HB10 gp160 for most peptides. The major role in recognition was ascribed to Asp-664, Lys-665, and Trp-666. The epitope-mimicking peptides were tested for immunogenicity. Antibodies to gp41 were detected in serum of immunized rabbits.

[Elaboration of live measles vaccine technology on human embryo lung diploid cell culture L-68]. / Razrabotka tekhnologii proizvodstva zhivoi korevoi vaktsiny na osnove kul'tury kletok émbriona cheloveka L-68.

Measles predominates among childhood droplet infections in many countries. Immunization of all human beings sensitive to this infection is the only radical measure in controlling measles. The quality of a vaccine is primarily determined by the properties of the virus strains and cell cultures and technology of production. Now live measles vaccine is produced in or country on the basis of fibroblasts from Japanese quail embryo. The production of live measles vaccine in the primary cell cultures has a number of drawbacks caused by the nonstandard pattern of the substrate and the probability of contamination. The use the certified human diploid cells deposited in liquid nitrogen in sufficient quantities is promising. The authors have elaborated a new technology of live measles vaccine production by using the Leningrad-16 virus strain on the basis of attested L-68 diploid cell culture from the human fetal lung. Experimental batches of vaccine were obtained and attested in accordance with the present requirements for immunobiological products.

[Enhancement of epidermal regeneration by recombinant vaccinia virus growth factor]. / Usilenie épidermal'noi regeneratsii s pomoshch'iu rekombinantnogo faktora rosta virusa ospovaktsiny.

Examining the specific activity has showed that recombinant vaccinia virus growth factor binds to appropriate receptors on the A-431 cell surface and prompts the healing acceleration of degree III burns in rats. This recombinant factor did not demonstrate pyrogenicity or toxicogenicity in tests on rabbits, guinea-pits, noninbred albino mice.

[Attempts to develop a vaccine against Ebola fever]. / Popytka polucheniia vaktsiny protiv likhoradki Ebola.

Data on the immunopathogenesis of Ebola fever in laboratory animals are presented and the efficacy of some methods of vaccine prophylaxis discussed. Antiviral immunity induced in guinea pigs by injection of inactivated viral agents did not protect them from infection, whereas injections of a nonlethal strain of the virus in ascending doses led to the formation of immunity preventing the development of disease upon inoculation with a lethal strain in high doses. The role of some viral peptides in the development of immune response is shown and variants of recombinant constructions for the prevention of Ebola fever are offered.