Risk of HIV Infection and Lethality Are Decreased in CCR5del32 Heterozygotes: Focus Nosocomial Infection Study and Meta-analysis.

CCR5del32 Homozygous deletion in the chemokine receptor R5 gene provides almost complete protection to individuals against HIV infection. However, data relating to the protective effect forCCR5del32 heterozygous individuals have been contradictory. The frequency of theCCR5del32allele in population control cohorts was compared with that of a group of children (27 Kalmyks and 50 Russians) infected by G-subtype HIV-1 in a nosocomial outbreak. The frequency of theCCR5del32allele was shown to be lower among the infected children in comparison with that of the control group; however, the difference was small and statistically insignificant. Similar results were obtained in a number of earlier studies. The insignificance of the small differences could be a result of one of two reasons. (i) The fact that there is no protective effect of the heterozygous state, and that the phenomenon depends only on the fluctuation of allele frequencies. In this case, there would be no differences even if the infected cohort is enlarged. (ii)The protective effect of the heterozygous state is real; however, the size of the studied cohort is insufficient to demonstrate it. In order to discern between these two reasons, a meta-analysis of data from 25 published articles (a total of 5,963 HIV-infected individuals and 5,048 individuals in the control group, including the authors' own data) was undertaken. A conclusion was drawn from the meta-analysis that theCCR5del32 allele protects individuals against the HIV infection even in a heterozygous state (OR=1.22, 95%CI=1.10-1.36). The risk of HIV infection forCCR5 wt/del32 heterozygotes was lower by at least 13% as compared to that for wild typeCCR5 wt/wthomozygotes. Prior to this study, no data of the type or any conclusions had been published for Caucasians. The mortality rate in the 15 years following the infection was found to be approximately 40% lower forCCR5del32 heterozygotes in comparison with that for the wild type homozygotes in the studied group. The size of the studied group was insufficient to claim difference validity (OR=2.0;p= 0.705), even though the effect quantitatively matched the published data. The features of the meta-analysis influencing the threshold level and the statistical validity of the effects are being discussed. The level of theCCR5del32 protective effect on the chances to be infected with HIV and on the outcome of the HIV infection was assessed for various ethnic groups.

[Analysis of HIV-1 integrase gene polymorphism in an HIV-infected population from the nosocomial outbreak of HIV infection in the south of Russia in 1989].

The paper presents the data of an investigation of the polymorphism of the pol gene encoding HIV-1 integrase in a HIV subtype G infected population formed during the 1989 HIV-infection outbreak. The investigators analyzed 41 samples of the viruses obtained in 2005-2007. Polymorphism at codons associated with integrase resistance to chemicals was observed in 11 virus variants. The circulation of mutation viruses that potentially promote the formation of resistance to the integrase inhibitors raltegravir and elvitegravir has been established in untreated patients.

[Investigation of genetic polymorphism of the integrase gene in the HIV-1 subtype A populations circulating in the Russian Federation].

The paper presents the data of an investigation of the genetic polymorphism of the pol gene encoding viral integrase (IN) in a HIV subtype A infected population in the Lipetsk Region. The investigators analyzed 32 virus subtype A samples obtained in 2002-2007. Polymorphism at the codons associated with IN resistance to chemicals was observed in 7 virus variants. The found substitutions had a pattern of genetic polymorphism and were unassociated with resistance in 6 patients with the test subtype A population. At the same time, minor RAL resistance mutation was revealed in 1 (3.1%) virus variant while the similar mutations in the subtype G population were about 10%.

[Molecular epidemiologic characteristics of HIV-1 variants isolated in the Lipetsk region].

Molecular epidemiologic study of HIV-1 variants isolated in the Lipetsk region during 1994 - 2006 period was performed. It has been shown that 3 env-subtypes (A, B, and C) and 3 gag-subtypes (A, B, and C) are widespread in the region. The virus was transmitted both sexually and by injecting drug users. Phylogenetic analysis of gag and env genes nucleotide sequences was performed, which revealed that 4 variants of HIV-1 with genotypes gagA/envA, gagB/envB, gagC/envC, and recombinant gagA/envB are circulating in the region.

[Genetic characteristics of HIV-1 subtype G reverse transcriptase gene].

The paper presents data of a study of the structure of reverse transcriptase gene in the population infected with HIV subtype G formed during the 1989 HIV infection outbreak in the North-Caucasian region. The authors analyzed 3 samples obtained in 1993-1994 and 17 samples taken in 2000-2001. The phylogenetic analysis indicated that polymerase of the test virus variants belonged to HIV-1 subtype G. The mutations occurring with azidothymidine therapy did not differ from those in subtype B. Analysis of the mutations of resistance to other nucleoside and non-nucleoside reverse transcriptase inhibitors did not show great differences from subtype B either.

[Prevalence of mutations of resistance to HIV-I reverse transcriptase inhibitors among HIV-infected patients in the Southern Federal District].

To study the regularities in the spread of drug-resistant HIV-1 strains among HIV-infected patients in the Southern Federal District (SFD), the HIV-1 pol gene site encoded for reverse transcriptase was sequenced in the samples taken from 22 HIV-infected SFD patients who received or did not receive antiretroviral therapy (ARVT). Analysis of the primary sequences of the HIV-1 pol gene in SFD patients untreated with antiviral agents revealed the absence of both primary and secondary mutations of resistance to a nucleoside reverse transcriptase inhibitor (NRTI) and a non-nucleoside reverse transcriptase inhibitor (NNRTI). The group of patients receiving antiviral treatment was found to have different drug resistance mutations in the HIV-1 pol gene: K70R, M184V, K219Q, T215Y/F, L74V, etc. Moreover, the patients on ARVT had higher CD4 T lymphocyte levels and higher immunoregulatory index in the presence of significantly lower HIV replication than the untreated patients. The authors make recommendations how to study HIV resistance in patients who are to be treated and are receiving ARVT and advise to monitor the spread of drug-resistant HIV strains in the SFD.

[Epidemiological characteristics of HIV infection in the Lipetsk Region in 1993-2005].

The spread routes of HIV infection in the Lipetsk Region showing a low incidence of this infection rate are analyzed in detail. The findings indicate that the trend in identification of HIV-infected persons in this region reflects the all-Russian HIV-infection spread situation, but the epidemic process has a number of features.

[Analysis of changes in the gp120 V3 region as observed in some patients with HIV infected from a common infection source]. / Analiz izmenchivosti oblasti V3 gp120 u otdel'nykh patsientov v kogorte VICH-infitsirovannykh, imeiushchikh edinyi istochnik zarazheniia.

HIV-1 genome regions encoding the gp120 V3 part were sequenced in samples isolated from persons belonging to the category of those infected in the Rostov-Elista outbreak and having the common infection source. Samples were obtained from 5 patients in 1992 and in 2001. A total of 27 sequences obtained in 1992 and 35 sequences obtained in 2001, 2 to 8 sequences for each patient, were analyzed. The diversity level of V3 sequences made, in some patients, 2.2% in 1992 and went up to 4.2% in 2001 samples (p < 0.07). The ratio between the synonymous and non-synonymous substitutions was determined for the gp120 V3 region. The mean ratio value made 0.12 in 1992 samples and 0.23 in 2001 samples. The obtained data confirm the assumption, made previously in a population analysis, on the evolution of the gp120 V3 epitope towards substitution of the Lg initial structure in positions 14 and 15 (through intermediate stages represented by the IG and FG structures) for the FA structure.

[HIV-1 serotyping of V3-mimicking peptides circulating in the Republic of Tajikistan among intravenous drug users]. / Serotipirovanie na osnove V3-imitiruiushchikh peptidov variantov VICH-1, tsirkuliruiushchikh v Respublike Tadzhikistan sredi lits, upotrebliaiushchikh narkotiki vnutrivenno.

The paper summarizes the results of HIV-1 serotyping by using 56 positive sera collected in the territory of the Republic of Tajikistan (RT) from intravenous drug users (IVDU). It was made by solid-phase ELISA based on synthetic peptides, mimicking different variants of the apical epitope of the HIV-1 gp120 V3-loop. Two types of conjugates, those specific to human IgG and IgA, were used to detect the immune complexes. Serotypes, as determined according to IgG and IgA-based ELISA, coincided, however, the latter were proven to be more suitable for serotyping. There is a high level of HIV-1 serotype heterogeneity among IVDU in RT; altogether, 4 serotypes were identified, i.e. B (10%), B+A/C (18%), A/C (20%) and A/C+B (52%). The modern serotype HIV-1 diversity in RT resembles the epidemiologic situation in the territory of the former USSR as observed in the late 80-ies-early 90-ies of the last century.

[A retrospective analysis of population type variability of the region V3 gp120 in HIV infected patients with a unified infection focus]. / Retrospektivnyi analiz populiatsionnoi izmenchivosti oblasti V3 gp120 v kogorte VICH-infitsirovannykh, imeiushchikh edinyi istochnik zarazheniia.

Regions of HIV-1 genome, encoding the V3 gp120 region, were sequenced in materials that were sampled from persons belonging to the category of individuals infected from a single source during the Rostov-Elista outbreak. The samples were obtained in 1991-1992 (10 pieces) and in 2000-2001 (16 pieces), which amounts to 8% and 13%, respectively, of a total number of patients infected by the beginning of 1990. It was established that the level of the population variability of amino-acid sequences in region V3 amounted, in 1991-1992, to 5.2% and increased to 9.1% in the samples of 2000-2001. A comparison of amino-acid sequences in region V3 from the collections of 1991-1992 and of 2000-2001 revealed the below amino-acid substitutions: from Ser13 to His, from Ley14 to Phe, from Phe to Ley and from Ley to Ala. An analysis of V3 B-epitope showed that the basic trend of its evolution consists in reciprocal transitions of L to F in position 14 and of F to L in position 20. Such substitutions ensure further changes at the "top" of the V3-loop from GPG to APG.

[Echinococcus granulosus genotyping by PCR-RFLP]. / Primenenie metoda otsenki polimorfizma dlin restriktsionnykh fragmentov produktov polimeraznoi tsepnoi reaktsii (PTSP-PDPF) dlia genomnogo tipirovaniia Echinococcus granulosus.

A method to assess the length polymorphism of restriction fragments of polymerase chain reaction (PCR-LPRF) has been developed and used for the genomic typing of E. granulosus. A scheme optimal under our conditions has been chosen for analysis. To test the chosen scheme, 10 E. granulosus samples from our collection made in Russia and Kazakhstan were examined. The material was obtained from 5 sheep, 1 dog, 2 cows, and 1 man. The E. granulosus genotypes detected in Russia and Kazakhstan do not differ from those described earlier.

Immunogenic properties of reverse transcriptase of HIV type 1 assessed by DNA and protein immunization of rabbits.

Genetic immunization may be one way to prime individuals for a subsequent broad anti-HIV-1 immune response. Reverse transcriptase of HIV-1 (RT) presents a selective target for attempts to arrest replication of HIV-1. Rabbits immunized with a plasmid carrying the gene for reverse transcriptase HIV-1 (RT DNA) developed potent antibody and cellular responses to the gene product. The immunogenic properties of RT DNA and recombinant reverse transcriptase were compared in rabbits. The specific immune responses were similar to those reported previously for HIV-1 infected humans. The array of B and T cell epitopes recognized in RT DNA-immunized rabbits was broader than in rabbits immunized with the recombinant RT. We localized seven novel B and T cell epitopes and concordance between B cell and helper T cell epitopes was observed. B cell epitopes of RT induced proliferation of peripheral blood mononuclear cells and were active as helper T cell epitopes. T cell-proliferative responses to the epitopes of RT preceded or paralleled the production of antibodies of the same specificity. Subdomains of reverse transcriptase involved in the enzymatic activity of RT were highly immunogenic. Anti-RT IgG partially inhibited reverse transcription in vitro.

Emergence of resistant variants of HIV in vivo during monotherapy with the proteinase inhibitor saquinavir.

We examined the phenotypic and genotypic properties of virus from the peripheral blood mononuclear cells (PBMC) and plasma of eight HIV-1-infected asymptomatic patients before and during monotherapy with the proteinase inhibitor saquinavir. Susceptibility of primary isolates to drug was assessed in PBMC culture by deriving IC50 and IC90 values. The observed increases in IC50 and IC90 after approximately one year of therapy with a dosage of 600 mg tds suggests the presence of virus resistant to saquinavir in vivo. The magnitude of this altered susceptibility ranged from three-fold to in one case 100-fold. In two patients a greater than eight-fold decrease in susceptibility to saquinavir was observed. Sequencing of the proteinase genes in viral RNA obtained from patient plasma and/or PBMC was carried out by PCR in parallel with sensitivity testing. In each case between nine and 12 clones were analysed. In the two patients from whom virus had greater than eight-fold reduction in susceptibility, a point mutation was observed in the viral proteinase (Leu90--> Met/Ile). Further mutations were observed at residues 36, 71 and 84 in these subjects. In a third patient, in whom an eight-fold increase in HIV IC50 of saquinavir was observed, no mutations were detected in the proteinase; sequencing of proteinase cleavage sites in viral gag-pol revealed no significant mutations. In no patient was a Gly48-->Val mutation observed, although this has been associated with resistance in vitro. The Leu90-->Met mutation was observed in five subjects, but a greater than eight-fold phenotypic change in antiviral susceptibility was seen in only two of these. Hence, in vivo, the Leu90-->Met but not the Gly48-->Val mutation is necessary, but not sufficient, for phenotypic resistance to saquinavir in HIV.

[New phospholipids--inhibitors of human immunodeficiency virus reproduction. Synthesis and antiviral activity]. / Novye fosfolipidy--ingibitory reproduktsii virusa immunodefitsita cheloveka. Sintez i antivirusnaia aktivnost'.

Several novel phosphatidic acid derivatives of 3'-azido-3'-deoxythymidine and 2',3'-dideoxyinosine were synthesized, which contained dialkylphosphatidyl, dialkylthiophosphatidyl moieties, as well as diacylphosphatidyl moiety with either 14,14,14-trifluoro-12-oxatetradecanoyl or natural acyl residues inherent in egg yolk phosphatidylcholine. Diacylphosphatidyl derivatives of glycyrrhetinic acid were also prepared. All the synthesized compounds exhibited significant anti-HIV activity. The glycyrrhetinic acid derivatives are of special interest because of their low toxicity and a supposedly different mechanism of action.

[A new oxa-analog of myristic acid, suppressing replication of the human immunodeficiency virus]. / Novyi oksaanalog miristinovoi kisloty, podavliaiushchii replikatsiiu virusa immunodefitsita cheloveka.

A series of oxaanalogs of myristic acid were synthesized and tested for antiviral activity in MT4 cells infected with human immunodeficiency virus 1 (HIV-1). The synthesized acids have no toxic effect on uninfected MT4 cells at a concentration of 100 microM. 14,14,14-Trifluoro-12-oxatetradecanoic acid substantially (by 75%) inhibits the reproduction of HIV-1. Other compounds synthesized, (7Z)-13-, (9Z)-13-, and (7Z)-11-oxatetradecenoic acids, exhibit no antiviral effect.

[An analysis of the blood sera of patients infected with HIV-1 for the presence of antibodies to the gp120 and gp41 sequences]. / ANaliz syvorotok krovi patsientov, infitsirovannykh VICh-1, na nalichie antitel k posledovatel'nostiam gp120 i gp41.

Blood sera, originating four regions of Russia and Byelorussia and previously tested for the content of antibodies to HIV-1 proteins, were studied in the enzyme immunoassay on the basis of recombinant sequences gp160, as well as on the basis of oligopeptides corresponding to sequences V3 of six HIV-1 strains. The possibility of using sequences gp160, contained in fusion polypeptides synthesized in Escherichia coli cells, for the detection of antibodies in laboratory research was shown. Differences in the reactivity of the sera under study with respect to fragments gp160 correlated with the geographical origin of these sera: similarity between the serum samples from Elista and Rostov and their difference from serum samples collected in other regions were shown.

[Detection of human immunodeficiency virus (HIV-1) p24 antigen in immune complexes]. / Vyiavlenie antigena p24 virusa immunodefitsita cheloveka (VICh-1) v sostave immunnykh kompleksov.

Enzyme-linked immunosorbent assay (ELISA) test-system for HIV-1 p24 antigen detection in human blood specimens has been developed. Use of complex biotin-streptavidin-horseradish peroxidase conjugate increased the sensitivity of the method to 50 pg/ml. The test system is characterized by 100 percent specificity, whereas direct p24 assay detects the antigen in only 5 percent of 100 sera of HIV-infected patients, since even low-titer sera efficiently mask p24. In order to extend the use of the test-system, destruction of immune complexes by different methods has been undertaken in experiments with different HIV-1 strains (RF, BRU). Acid hydrolysis detected the antigen in 30% of samples, appreciably improving the prognostic value of the test-system. A manifest correlation was revealed between the frequency of p24 detection in the blood plasma and clinical diagnosis.

[Synthesis and comparative analysis of antigenic activity of peptides from hypervariable region V3 of human immunodeficiency virus I gp120 surface glycoprotein]. / Sintez i sravnitel'nyi analiz antigennoi aktivnosti peptidov gipervariable'noi oblasti V3 poverkhnostnogo glikoproteida gp120 virusa immunodefitsita cheloveka tipa I.

A set of peptides (amino acid positions 10-23) corresponding to seven most widely spread variants of the gp120 V3 domain in the HIV-infected population of South Russia were prepared by the solid-phase synthesis. A laboratory variant of the indirect enzyme-linked immunosorbent assay (ELISA) was developed for the determination of V3 specific antibodies with use of the peptides synthesized. The analysis of the V3-specific antibodies in HIV-infected using the elaborated test-system revealed a correlation between the V3 variants distribution and the occurrence of antibodies against these variants.

[Expression of the gene for the gp46 surface glycoprotein from the human T-cell leukemia virus type I (HTLV-I) in bacteria]. / Ekspressiia gena poverkhnostnogo glikoproeida gp46 virusa T-kletochnogo leikoza cheloveka tipa I (HTLV-I) v bakteriiakh.

A set of recombinant plasmids containing different fragments of HTLV-I env gene has been constructed on the basis of pUR290-pUR292 vectors. The hybrid proteins containing different fragments of ENV predecessor in the C-terminal of beta-galactosidase differed in stability in Escherichia coli cells. The presence of N-terminal of ENV predecessor in recombinant proteins considerably decreases their resistance to proteases of the bacterial cell. Elimination of this fragment led to obtaining of the recombinant plasmid pESG coding for the high level of synthesis of the env-specific hybrid polypeptide (up to 30% of the total cellular protein). This 134 Kda protein is able to interact efficiently with the HTLV-I positive sera and may be used in the diagnostic test-systems for identification of the HTLV-I infected patients.

Molecular epidemiology of HIV-1 in the former Soviet Union: analysis of env V3 sequences and their correlation with epidemiologic data.

OBJECTIVE: To investigate the HIV-1 V3 sequence diversity in the former Soviet Union in 30 subjects infected with HIV-1 via different modes of transmission. PATIENTS: A cohort of children infected after exposure to nonsterile needles during the epidemic in 1988-1989 in southern Russia (Elista, n = 12 and Rostov-on-Don, n = 10), and eight HIV-seropositive subjects from Belarus (Minsk), infected via sexual (n = 7) and parenteral (n = 1) infection. METHODS: The HIV-1 V3 encoding region was amplified by nested polymerase chain reaction on DNA of primary peripheral blood mononuclear cells collected from the study subjects and then cloned and sequenced. RESULTS: The alignment of 127 V3 sequences from 22 patients in the cohort group demonstrated common consensus sequences in both the Elista and Rostov samples. The average means of interperson variation were 5.9 and 6.6% in Elista and Rostov subjects, respectively, and comparable to the mean intraperson variation. The average mean interperson variation between nucleotide sequences of HIV patients infected through sexual transmission was considerably higher (14.9%). CONCLUSION: V3 sequence analysis confirms the epidemiologic data which support the transmission of HIV-1 in children from a single source, and suggests the infection of a mother from her parenterally infected child. Furthermore, the genetic variability of HIV-1 V3 in the noncohort group was particularly divergent indicating the heterogeneity of the virus circulating in the former Soviet Union.

PIP: In 1988, an HIV-1 epidemic occurred in Elista, Kalmyk Republic, Russia, among 90 children in two hospitals after exposure to blood contaminated needles from an HIV infected infant. A few months later, a similar HIV-1 outbreak in children occurred in Rostov-on-Don, Russia, probably a result of transporting children from Elista to Rostov-on-Don hospitals. In Rostov-on-Don, it appears that seven HIV infected infants transmitted HIV to their mothers during breast feeding. Health workers collected blood samples from 22 HIV-1 infected subjects in Elista (n = 12) and Rostov-on-Don (n = 10 including 1 mother-child pair) and from 8 control subjects who became infected with HIV-1 via sexual (7) and parenteral (1) transmission from Minsk, Belarus. Researchers wanted to determine the extent of the diversity of proviral DNA encoding the V3 loop from different patients in the children cohort. They used nested polymerase chain reaction on DNA of primary peripheral blood mononuclear cells and then cloned and sequenced them to detail the HIV-1 V3 encoding region. The Elista and Rostov-on-Don samples shared common consensus sequences (127 nucleotide sequences) in the V3 region. The average mean interperson variation between the nucleotide sequences of HIV patients infected through sexual transmission from Minsk was 14.9%, which was much higher than those for Elista and Rostov HIV patients infected through parenteral transmission (5.9% and 6.6%, respectively). The major nucleotide sequence in the mother in the Rostov group, who was presumably infected with HIV by her HIV infected infant during breast feeding, matched that of her daughter. The mother had no history of blood transfusion or any other risk factors except breast feeding. These findings confirm that the Elista and Rostov groups shared a common HIV source. They also suggest that breast feeding was the route of HIV transmission for the mother. The genetic variability of HIV-1 V3 in the control group demonstrated the heterogeneity of HIV-1 in the former USSR.