RESUMO
Action of three nucleoside analogues with azido-group on AIDS virus (human immunodeficiency virus--HIV) reproduction was studied in two cell species. Among these compounds there were 3'-azido-3'-deoxythymidine (Az-T), 3'-azido-3'-deoxyarabinothymidine (Az-AT) and 3'-azido-2',3'-dideoxy-5-methylcytidine (Az-dCMe). All compounds prevent cells against HIV infection, but Az-T action was expressed more strongly. The reason for the lower activity of the Az-AT and Az-dCMe bases is probably stipulated by their poor conversion to the corresponding 5'-triphosphates. Az-T 5'-triphosphate blocks HIV reproduction due to the inhibition of the viral reverse transcriptase.
Assuntos
Azidas/farmacologia , Didesoxinucleosídeos/farmacologia , HIV/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Zalcitabina/análogos & derivados , Zidovudina/análogos & derivados , Zidovudina/farmacologia , Células Cultivadas , HIV/enzimologia , HIV/fisiologia , Humanos , Linfócitos/microbiologia , Proteínas dos Retroviridae/biossíntese , Inibidores da Transcriptase Reversa , Ribavirina/farmacologiaRESUMO
The antiviral activity of 3'-azido-2',3'-dideoxynucleoside 5'-phosphate analogues: 5'-phosphonomethylene-3'-azido-2',3'-dideoxythymidine, 5'-methylphosphonate and 5'-phosphite of 3'-azido-2',3'-dideoxythymidine, 5'-phosphites of 3'-azido-2',3'-dideoxyadenosine and guanosine was investigated in HIV-infected cell cultures (human lymphoblastoid cells). The effectivity of inhibition of HIV-reproduction in cells by these substances was close or even higher than that for the corresponding 3'-azido-2',3'-dideoxynucleosides, whereas their toxicity was lower than that of nucleosides. These substances are supposed to be transported into the cells and to be transformed into the corresponding 5'-triphosphate analogues under the action of cell kinases. It is possible that such agents are terminator substrates of virus reverse transcriptases and thus inhibit the biosynthesis of DNA chains.
Assuntos
Antivirais , Azidas/farmacologia , Didesoxinucleosídeos/farmacologia , HIV-1/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Antivirais/síntese química , Azidas/síntese química , Células Cultivadas , Fenômenos Químicos , Química , Didesoxiadenosina/análogos & derivados , Didesoxiadenosina/síntese química , Didesoxiadenosina/farmacologia , Didesoxinucleosídeos/síntese química , HIV-1/fisiologia , Humanos , Organofosfonatos/síntese química , Organofosfonatos/farmacologia , Zidovudina/análogos & derivados , Zidovudina/síntese química , Zidovudina/farmacologiaRESUMO
3'-Mercapto-3'-deoxy-TTP was synthesized and tested as DNA chain terminator nucleotide for calf thymus alpha DNA polymerase, E. coli DNA polymerase I (Klenow fragment), terminal deoxyribonucleotidyltransferase (Bollum enzyme) and reverse transcriptase from AMV- and HIV-I-viruses. It was shown that the compound terminates DNA chain elongation by reverse transcriptases selectively and irreversibly. Other tested DNA polymerases do not use this nucleotide analogue as a substrate. 3'-Mercapto-3'-deoxythymidine was tested on lymphoblastoid T-cell line MT-4 with HIV-viruses and shown to suppress viruses as efficiently as 3'-azido-3'-deoxythymidine.
Assuntos
DNA Polimerase I/metabolismo , DNA/biossíntese , Nucleotídeos de Timina/farmacologia , Animais , Vírus da Mieloblastose Aviária/enzimologia , Sequência de Bases , Catálise , Bovinos , Linhagem Celular , DNA/efeitos dos fármacos , Eletroforese em Gel de Ágar , Escherichia coli/enzimologia , HIV-1/enzimologia , Dados de Sequência Molecular , Inibidores da Transcriptase ReversaRESUMO
Trials of clinically advantageous national inducers, thymus hormones, as well as human recombinant alpha 2-interferon were carried out in cultures of continuous lymphoblastoid cells H9/IIIB infected with HIV virus. The virus-inhibiting effect for HIV was observed with interferon in doses of 10-100 IU/ml. At a concentration of interferon of 1000 IU/ml, virus replication was inhibited completely, the interferon doses used exerting no marked toxic or antiproliferative effect on the cells. Human interferon inducers, poly(G).poly(C), PXL-6, dsRNA in concentrations of 50-100 micrograms/ml inhibited virus reproduction significantly. The highest antiviral effect was achieved with yeast dsRNA. The preparations of immunomodulators, thymarin, the 5th and 7th fractions of thymosin, noticeably stimulated proliferation of infected T-lymphocytes, reducing the relative number of cells carrying the virus-specific antigen. Combined use of preparations with different mechanisms of the antiviral effect may be advantageous in prevention and treatment of AIDS.
Assuntos
Antivirais/farmacologia , HIV/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Depressão Química , HIV/fisiologia , Indutores de Interferon/farmacologia , Interferon Tipo I/farmacologia , Compostos Orgânicos , Poli C/farmacologia , Poli G/farmacologia , Proteínas Recombinantes/farmacologia , Timosina/análogos & derivados , Timosina/farmacologia , Hormônios do Timo/farmacologiaRESUMO
Treatment of AIDS is a complicated problem since there is a limited set of preparations tested and most of them appear to be ineffective. Their main target is HIV reverse transcriptase. The data presented show that proteolysis inhibitors are able to inhibit HIV reproduction in cell culture. The compounds used in this study, gordox and kontrikal, are officinal drugs and therefore it is worthwhile to study their effect in patients with AIDS.
Assuntos
Aprotinina/farmacologia , HIV/efeitos dos fármacos , Inibidores de Proteases/farmacologia , Inibidores da Tripsina/farmacologia , Replicação Viral/efeitos dos fármacos , Células Cultivadas , HIV/fisiologiaRESUMO
The possibility of optimizing the conditions for the cultivation of cells producing human immunodeficiency virus (HIV) was explored. The stimulating effect on the cell cultures of interleukin-2 and specific anti-interferon antibodies was examined. The individual use of interleukin-2 or anti-interferon antibody preparations did not result in any marked enhancement of HIV virus reproduction in the cells, whereas combining of interleukin-2 which stimulated proliferation of T-lymphocytes with poly- and monoclonal anti-interferon antibodies proved to be effective increasing the expression of virus-specific antigens in the cells 1.5-fold. It seems expedient to carry out further screening of different reagents and combinations thereof capable of significantly increasing HIV virus reproduction in cell cultures which would serve as the antigen for diagnostic systems.
Assuntos
Fibroblastos/microbiologia , HIV/crescimento & desenvolvimento , Linfócitos/microbiologia , Anticorpos/análise , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos , Linhagem Celular , Fibroblastos/efeitos dos fármacos , Fibroblastos/imunologia , HIV/imunologia , Humanos , Interferons/imunologia , Interleucina-2/farmacologia , Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Estimulação Química , Cultura de Vírus/métodos , Replicação Viral/efeitos dos fármacosRESUMO
The paper deals with further search for substances modifying metabolic processes, changing the conditions of virus existence in the host cell and, consequently, possessing a certain antiviral activity. The antiviral effects of aromatic carbonic acids amides including trisubstituted benzamides and nicotinamide were tested. Five out of 8 substances tested possessed antiviral activity which might be due to their capacity to inhibit the activity of the enzymes of ADP-ribosylation. By blocking cellular processes of ADP-ribosylation, the substances under study depressed DNA capacity for reparation, inhibited differentiation and transformation of cells, i.e. had an indirect effect on reproduction of viruses. The universal nature of the processes coursing with participation of NAD(+)-dependent ADP-ribosylation suggests that the range of antiviral activity of inhibitors of mono- and poly-ADP-ribosylation would not be limited to HIV infection but would be rather wide.
Assuntos
Antivirais/uso terapêutico , Infecções por HIV/tratamento farmacológico , HIV-1 , Inibidores de Poli(ADP-Ribose) Polimerases , Difosfato de Adenosina/metabolismo , Adenosina Difosfato Ribose/antagonistas & inibidores , Células Cultivadas , Efeito Citopatogênico Viral/efeitos dos fármacos , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , HIV-1/efeitos dos fármacos , HIV-1/patogenicidade , Humanos , Niacinamida/uso terapêutico , Zidovudina/uso terapêuticoRESUMO
Cell systems infected with 63 strains of types 1 and 2 HIV virus (HIV-1 and HIV-2) were examined under electron microscope. HIV virions were most frequently detected near the cell membrane or budding from it. In the cytoplasm HIV occurred only in vacuole-like formations. Accumulations of mature virions were seen in the cell-to-cell space. Mature particles of HIV-1 and HIV-2 differed by their morphology from oncoviral C particles and were similar rater to the Visna/Medi type Lentiviruses. Morphological analysis of HIV strains isolated in Russia demonstrated their similarity to be foreign HIV strains.
Assuntos
HIV-1/isolamento & purificação , HIV-2/isolamento & purificação , Linhagem Celular/ultraestrutura , Linhagem Celular/virologia , Humanos , Microscopia Eletrônica , Vírion/ultraestruturaRESUMO
Eight patients with type I diabetes mellitus (D-I), seven patients with type II diabetes mellitus (D-11) and 8 healthy donors were examined. The disease standing did not exceed 1 year since the moment of the diagnosis establishment. The patients with D-I manifested activation of natural killers (NK) as compared to their activity in the donors and patients with D-II (76.05 +/- 6.5%, 52.33 +/- 9.55% and 55.39 +/- 10.63%, respectively, p less than 0.01) in the presence of the attenuated response of NK to interleukin-2 and alpha-interferon, determined by NK prestimulation. The amount of NK (CD16-positive) in D-I was significantly less than in the donors and patients with D-II. The high activity of NK in D-I correlated with an increase of receptor expression for transferrin on the mononuclear cells of peripheral blood. At the same time 5 out of the 8 patients with D-I and 2 patients with D-II out of the 7 demonstrated the rise of serum alpha-interferon (in the titer 1:40 and over). Activation of NK and the rise of serum interferon may be due to viral etiology of the disease and may play a role in the autoimmune process in patients suffering from D-I.
Assuntos
Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 2/imunologia , Células Matadoras Naturais/imunologia , Adolescente , Adulto , Testes Imunológicos de Citotoxicidade/métodos , Diabetes Mellitus Tipo 1/etiologia , Diabetes Mellitus Tipo 2/etiologia , Feminino , Humanos , Interferon Tipo I/farmacologia , Interleucina-2/farmacologia , Células Matadoras Naturais/efeitos dos fármacos , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Proteínas Recombinantes/farmacologia , Linfócitos T Citotóxicos/efeitos dos fármacos , Linfócitos T Citotóxicos/imunologiaRESUMO
A study was made of the level of anti-HTLV-111 using an enzyme immunoassay (Emzygnost-anti-HTLV-111) in 75 donors and 143 patients with various diseases including SLE (57), rheumatoid arthritis (9), Sjögren's disease (3), Bekhterev's disease (2), polymyositis (2), dilatation cardiomyopathy (30), and acute lymphocytic leukemia (40). Anti-HTLV-111 were detected in 26 SLE patients and in one patient with polymyositis. A study of positive sera using immunoblotting showed no interaction with virus antigens. A conclusion was made that the detection of anti-HTLV-111 in systemic rheumatoid diseases was a false-positive result.
Assuntos
Anticorpos Antivirais/análise , Reações Antígeno-Anticorpo , Antígenos Virais/imunologia , HIV/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Anticorpos Antivirais/imunologia , Doadores de Sangue , Ensaio de Imunoadsorção Enzimática , Reações Falso-Positivas , Humanos , Imunoensaio , Leucemia Linfoide/imunologia , Doenças Reumáticas/imunologiaAssuntos
Síndrome da Imunodeficiência Adquirida/diagnóstico , Reações Antígeno-Anticorpo , Anticorpos Anti-HIV/análise , Antígenos HIV/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Adolescente , Adulto , Complexo Antígeno-Anticorpo/análise , Doenças Autoimunes/imunologia , Reações Falso-Positivas , Feminino , Humanos , Testes Imunológicos/métodos , Masculino , Doenças Reumáticas/imunologiaRESUMO
A karyological analysis of twenty-two variants of eight cell lines, which differed in their susceptibility to the human immunodeficiency virus (HIV) and which had been obtained from different sources, was carried out by means of differential chromosome staining for G and C bands. The karyotypes of eight T-lymphoblastoid cell lines were identified, including five (MT-4, Molt-3, CEM, H-9, and Hut-78) not previously studied by cytogenetic methods. Karyotyping confirmed the identity of seventeen variants of the eight cell lines, and five variants of four lines were found to be misidentified. Comparative analysis of the cytogenetic characteristics of the three CEM-line variants demonstrates the need for karyotype evaluation in the course of in vitro cell cultivation. Fourteen identical marker chromosomes were revealed in H-9 and Hut-78 cell karyotypes, confirming the common origin of these two lines. It was found that the cells of the HIV-susceptible lines had a tendency to undergo polyploidisation both during the initial stages after isolation and in the course of cultivation.