[The molecular and virological specificities of the epidemic of HIV infections in Russia and other CIS countries].
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Biomedical subjects
Publications and source records attributed to E V Kazennova.
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The frequencies of two mutations associated with the development of clinical symptoms upon infection with human immunodeficiency virus type 1 (HIV-1) were determined in a cohort of individuals from Moscow. Allelic frequency of the first mutation, CCR2-64I, causing the substitution of valine with isoleucine in the CCR2 chemokine receptor, was 0.1106 (95% confidence interval, 0.0714-0.1498). The frequency of the second mutation the G to A substitution in the 3'-untranslated region of the stromal-derived factor 1 encoding gene, SDF1-3'A, was 0.2125 (95% confidential interval, 0.1608-0.2642). Both values were slightly higher than those obtained earlier for Western European countries. This result can be explained by higher proportion of Asian immigrants, characterized by higher frequencies of these mutations, in the population of Moscow.
The paper presents data on the variants of human immunodefficiency virus type 1 (HIV-1) currently circulating in Russia. The subtype A HIV-1 variant dominating is shown to be most widespread among drug-injected users in the most regions under study. By using the results of an analysis of 1,464 blood samples taken in the past 4 years in 69 subjects of the Russian Federation, the authors have estimated that this HIV-1 variant is responsible for 93% of all HIV-infection cases in the country. The greatest regional genetic diversity was observed in Moscow and its mean (2.35(1.59) was found to be comparable to that (2.41(1.85) in the whole country. Penetration of the subtype A IV-1 variant early detectable among drug-users into other risk groups was noted.
The aim of this study was to develop and evaluate a simple V3 peptide-based enzyme immunoassay (PEIA) for large-scale serotyping of HIV-1 specimens derived from injecting drug users (IDUs) in the Russian Federation. Two synthetic peptides were evaluated, named P1 (RKSIHIGPGRAFYATGD) and P2 (RTSVRIGPGQVFYKTGD), in an PEIA on 63 HIV-1 IDUs sera for which genotypes had been determined by heteroduplex mobility assay (HMA) and sequencing. The sensitivities of P1 (subtype B) and P2 (subtype A) were 87% and 75% respectively. Specificity of the assay was 100% for both peptides, with 100% predictive values of a monoreactive positive test for both peptides. Using the PEIA with peptides P1 and P2, we have serotyped 375 of 477 serum samples derived from IDUs in 4 main sites of the HIV-1 epidemic in Russia. The results demonstrated a high level of subtype homogeneity in all regions studied. In 3 of 4 territories, Tver' (n=345), and Rostov-on-Don (n=61) regions, and Krasnodar Kray (n=27), 100% of typable sera were found to belong to env subtype A. On the other hand, all specimens serotyped in the Kaliningrad region (n=38) belonged to env subtype B, and there is strong evidence that the recombinant gagAenvB virus which has caused the largest outbreak of HIV-1 in Russia is located in this region. At the present time another parental strain with gagBenvB genotype is of minor importance in the IDUs HIV-1 epidemic in Russia.
Designing of recombinant plasmids pSB2 and pSB3 with the 932 bp HTLV-II env gene inserts encoding the full-length surface membrane glycoprotein gp46 is described. Vectors pGOmpF and pET32a expressing genes cloned under control of the late bacteriophage T7 promoter were used. Western blot analysis of cellular proteins derived from E. coli B834/pSB2 and E. coli B834/pSB3 revealed that 34 kD and 31 kD polypeptides corresponding to the full-length gp46 and its processed form without signal peptide were synthesized under control of these recombinant plasmids. Cytotoxic activity of the recombinant proteins towards bacterial cells was demonstrated. Both polypeptides specifically reacted to sera from humans infected with HTLV-II. High antigenic specificity of P34-HTLV-II proteins makes a promising candidate for diagnostic confirmation tests.
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A post-embedding technique for immunocytochemical analysis at the ultrastructural level was used to detect and localize HIV antigens on ultrathin sections of Lowicryl-embedded HIV-infected cells. A genomic probe containing ribosomal sequences and labeled with biotin was used to hybridize rRNA molecules in sections of animal cells embedded in Lowicryl. The method presently described offers the possibility to detect rapidly and precisely ribosomal gene expression and viral proteins at the ultrastructural level.
The results of cloning Corynebacterium diphtheriae phi 984 tox gene and its A and B subunits in Escherichia coli are presented. Regulatory sequences of tox gene are capable to promote effective expression in E. coli cells. A set of recombinant plasmids has been obtained which can determine the synthesis of A and B individual subunits and are suitable for constructing immunotoxins by gene engineering. The diphtheria toxin of 62 kDa synthesized in E. coli has enzymatic activity and reacts with antitoxin sera. Some sites for E. coli proteases are present in tox-specific polypeptides.
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Hybrid plasmids containing B. pertussis DNA insertions have been constructed with the use of the vector plasmid pBR 322 and the Pst I fragments of B. pertussis DNA. Some properties of the hybrid plasmids are characterized. The possibility of the expression of B. pertussis genes in the protein-synthetizing cell-free system obtained from E. coli has been demonstrated.
New restriction endonuclease has been isolated from Bordetella pertussis vaccine strain 305 and purified in 1 stage on Sepharose covalently bound with blue dextran. The isolated restrictase has been found capable of breaking down lambda-phag DNA into 7 fragments. According to its specificity, Bpe I is the isoschizomer of Hind III obtained from Haemophilus influenzae strain Rd.
Using ion-exchange chromatography on DEAE-cellulose and gel filtration, a purified NADH-dehydrogenase was prepared from cell membranes of A. laidlawii. The enzyme specific activity exceeds the activity of the membrane preparation 45-fold. The enzyme is thermolabile and sensitive to lecithine. Electrophoresis of the enzyme preparation in the presence of Na-DS resulted in two proteins with Mr of 65 000 and 72 000. Specific staining of the gels with triphenyltetrasolium chloride in the presence of Triton X-100 revealed two protein bands, while in the presence of Triton X-100 and urea only one protein band was produced.
The subtyping of 350 isolates of HIV-1, isolated on the territories of 38 subjects of the Russian Federation, was carried out. The analysis was made by the method of the comparative heteroduplex mobility assay, as well as by the determination of the sequence of genes env [correction of ens] (gp 120) and gag (p17-p24). The study revealed that more than 50% of all cases of HIV-1 infection were caused by closely related variants of subtype A virus. The number of cases of HIV-1 infection caused by recombinant virus A/B was not less than 25%. The total number of cases caused by viruses of subtypes C, D, E, F and H was not more than 5%.
Three types of recombinant human immunodeficiency virus have been found to circulate in Russia. The first type of the virus belongs to genotype gagAenvB, the second type belongs to genotype gagDenvG and the third type, to genotype gagAenvE. The virus of genotype gagAenvB circUlates in the population of drug addicts simultaneously with its "parent" viruses of genotypes gagAenvA and gagBenvB. The recombinant variant gagDenvG has African origin. The recombinant variant gagAenvE has probably been imported to Russia from South-East Asia.
Gag/env nucleotide sequences of human immunodeficiency virus type 1 (HIV-1) variants detected in drug users in Russia, Ukraine, and Belarus are analyzed. Two HIV-1 subtypes A and B circulate in this risk group. Genetic variability within one subtype is no higher than 3.1 and 3.9% for gag and env genes, respectively, suggesting the same source of infection in populations of drug users infected with the same subtype. Recombinant viruses with gagAenvB genotype, genetically related to parental strains of subtypes A and B, circulate in this risk group. This is the first report about HIV-1 recombinant of the two subtypes, for which both parental strains are known, directly confirming the in vivo recombination between different subtypes.
A serological test has been developed, allowing a rapid and reliable differentiation between envA and envB genotypes in the sera from HIV-1-infected IDUs and their sexual partners. We analyzed 556 specimens from HIV-1-seropositive IDUs and their sexual partners. The sera were collected in five regions of Russia with the greatest number of HIV-1-infected IDUs. High homogeneity of HIV-1 in each of the regions was observed. In Tver, Rostov-on-Don, Nizhny Novgorod, and Krasnodar 100% sera tested belonged to the env subtype A. All the patients from the Kalinigrad region were infected with the env subtype B virus, which is apparently a recombinant.
The paper sums up molecular epidemiological data on all variants of human immunodeficiency virus type 1 (HIV-1) isolated in the Perm region in August 1988-March 1998. A total of 24 foci (40 cases) of HIV infection were detected; 20 of these were imported from other regions (Russian Federation, Ukraine, Belarus, Bulgaria, Poland, Uzbekistan, Yugoslavia). In August 1988-September 1996 viruses of 3 env subtypes (B, D, and G) were isolated in the region, and the virus was mainly sexually transmitted. All cases of HIV infection recorded in September 1996-March 1998 were detected in intravenous drug users and subjects who had sexual intercourse with HIV-infected drug users. The same 3 variants of HIV-1 with genotypes gagAenvA, gagBenvB, and gagAenvB were isolated in the Perm region during this period as in other regions of Russia.
The genetic analysis of the variants of human immunodeficiency virus of type 1 (HIV-1), circulating among drug addicts in Moscow and Moscow Province, has been carried out. The serological analysis of 122 blood specimens taken from HIV-infected drug addicts, residing in Moscow and 22 settlements of the Moscow region, has shown that in this region HIV-1 variant of subtype A spreads among drug addicts. These data have been confirmed by the results of the analysis of 44 specimens, made with the use of the method of the heteroduplex mobility assay for gene env. As revealed in this study, HIV-1 variants spreading at present among drug addicts in Moscow and the Moscow region are genetically related to viruses of subtype A, detected earlier in this group of risk in other regions of Russia, the Ukraine, Belarus and other countries of Eastern Europe.