ABSTRACT
Previous studies have demonstrated that infection with HIV-1 clades might differentially contribute to the neuropathogenesis of HIV-1-associated dementia (HAD). HIV-1 transactivator regulatory protein (Tat) plays a major role in the process of disruption of neuronal function. It is not well understood how these HIV-1 subtypes exert different neuropathogenic effects. Activation of indoleamine-2,3-dioxygenase (IDO), the rate-limiting enzyme of the kynurenine pathway, leads to increased tryptophan catabolism and the generation of neurotoxins such as kynurenine (KYN). It is known that KYN plays a crucial role in the neuropathogenesis of HAD. We hypothesize that HIV-1 clade B and C Tat proteins might exert differential effects on human primary astrocytes by the upregulation of the IDO gene and protein expression as well as its activity and production of the neurotoxin KYN. RNA extracted from human primary astrocytes treated with either HIV-1 clade B and C Tat proteins was reverse transcribed and analyzed by quantitative real-time PCR to determine IDO gene expression. In addition, the enzymatic activity of IDO and the concentration of KYN were measured in cell lysates and culture supernatants. Our results indicate that HIV-1 clade B Tat protein significantly upregulated the IDO gene and protein expression, IDO enzyme activity, as well as KYN concentration compared to HIV-1 clade C Tat protein. Thus, our studies for the first time demonstrate that HIV-1 clade B Tat protein in human primary astrocytes appears to increase the level of neuropathogenic agents, such as IDO and KYN, as compared to HIV-1 clade C Tat protein. These results provide further evidence that the prevalence of HAD may be correlated with the difference in clades of HIV-1.
Subject(s)
Astrocytes/virology , Gene Expression Regulation , Gene Products, tat/physiology , HIV-1/physiology , Host-Pathogen Interactions , Indoleamine-Pyrrole 2,3,-Dioxygenase/biosynthesis , Astrocytes/chemistry , Cells, Cultured , Gene Expression Profiling , Humans , Kynurenine/analysis , Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
OBJECTIVES: The aim of our investigation was to study the HIV-1 env V3 loop sequences of the subtype A variant of the virus actively circulating in the territory of Eastern Europe (A1-EE). DESIGN: It appears that the characteristics of HIV-1 V3 loop sequences of the thoroughly studied subtype B strains determine the viruses tropism and phenotype. We were interested to find out to what extent these trends are preserved with the subtype A isolates spread in Eastern Europe. METHODS: About 474 HIV-1 strains were isolated by cocultivation with phytohemagglutinin stimulated and interleukin-2 activated PBMC. Isolates were subtyped after distinct regions gag, pol and env were amplified by nested polymerase chain reaction. V3 sequences were studied in detail and compared with the isolates that appeared as SI or NSI in test-cultures MT-2. RESULTS: The subtype A HIV-1 strains spread in Eastern Europe appeared to have V3 loop sequences with a high level of conservatism. The consensus sequences of the SI and NSI isolates proved identical, and correspond to the phenotype markers of subtype B NSI strains. Nevertheless, the subtype A SI isolates induced active syncytium-formation processes in the MT-2 test culture. CONCLUSION: The data obtained evidenced to the following: the HIV-1 subtype A variant spread in Eastern Europe, is capable of using for interaction with receptor structures of sensitive cells the tools and mechanisms, which are significantly different from those described for subtype B.
Subject(s)
HIV Envelope Protein gp120/chemistry , HIV Envelope Protein gp120/genetics , HIV Infections/virology , HIV-1/chemistry , HIV-1/genetics , Peptide Fragments/chemistry , Peptide Fragments/genetics , Amino Acid Motifs/genetics , Amino Acid Sequence , Base Sequence , Consensus Sequence , Epitopes/chemistry , Epitopes/genetics , Europe, Eastern/epidemiology , Genetic Variation , HIV Infections/epidemiology , Humans , Molecular Epidemiology , Molecular Sequence Data , PhylogenyABSTRACT
The investigation was outlined to study antibodies against some antigens of extracellular microbes associated with inflammation in broncho-pulmonary system and accessory nasal sinus - Staphylococcus aureus, Streptococcus pneumoniae, Branhamella catarrhalis - in individuals (18 patients) with different stages of HIV-infection. The level of antibodies was measured by ELISA and their Ab affinity was assessed by sodium thiocyanate-induced alteration of antibody-antigen interaction. To determine interrelations between antibody production and CD4(+) T lymphocyte number flow cytometry was employed. At the early stages of HIV-infection the levels of antibodies against Streptococcus pneumoniae and GMGM decreased, in comparison with HIV-negative donors. During HIV-infection course levels of antibodies against Staphylococcus aureus peptidoglycan, its antigen determinants and Streptococcus pneumoniae somatic antigen increased. Time affinity of antibodies against these antigens decreased. At all stages of HIV-infection and at all forms of its complications, we observed an increase of titer of antibodies to GMDP, antigenic determinant of peptidoglycan, which carried immunostimulating and adjuvant activities. HIV patients with CD4(+) T lymphocyte number <200 cells/&mgr;l displayed higher level of antibodies to bacterial antigens than that in patients with CD4(+) T lymphocyte number 200-400 per ml. The development of humoral immune response against some of extracellular bacteria is characterized, on the one hand, by their increased levels, and on the other hand, decreased affinity.
