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1.
Vopr Virusol ; 65(6): 307-316, 2021 Jan 07.
Artigo em Russo | MEDLINE | ID: mdl-33533227

RESUMO

Studying the regulation of signaling reactions of innate immunity by the hepatitis C virus (HCV) will help to reveal the causes of the transition of the acute form of the disease to a chronic course. The molecular mechanisms of activation by HCV RNA of innate immunity receptors TLR and RLR and signal transduction processes leading to the synthesis of IFN and inflammatory cytokines are considered. The inhibitory effects of non-structural and structural HCV proteins on immune signaling reactions are analyzed in detail. The information presented is the result of an analysis of literature data published in international databases mainly over the past 5 years. In conclusion, signaling receptors are proposed as targets for the development of new antiviral drugs with immunotherapeutic activity.


Assuntos
Hepacivirus/genética , Hepatite C/genética , Imunidade Inata/efeitos dos fármacos , RNA Viral/imunologia , Citocinas/genética , Hepacivirus/imunologia , Hepacivirus/patogenicidade , Hepatite C/imunologia , Hepatite C/virologia , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Humanos , Imunidade Inata/genética , Interferons/genética , RNA Viral/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Receptores Toll-Like/genética , Replicação Viral/efeitos dos fármacos , Replicação Viral/genética
2.
Bull Exp Biol Med ; 164(5): 636-640, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29577197

RESUMO

In culture of THP-1 cells differentiated into macrophages with PMA (THP-PMA macrophages) infected with influenza viruses of subtypes H1, H5 and H9, we measured the expression of TLR7 and RIG1 receptor genes, sensors of viral RNA and ribonucleoprotein, and the levels of production of inflammatory cytokines IL-1ß, TNFα, IL-10, and IFNα. The sensitivity and inflammatory response of THP-PMA macrophages to pandemic influenza A virus H1N1pdm09 and avian influenza H5N2 and H9N2 viruses correlate with the intracellular level of their viral RNA and activation of the RIG1 gene. Abortive infection is accompanied by intensive macrophage secretion of TNFα, IL-1ß, and toxic factors inducing cell death. Activity of endosomal TLR7 receptor gene changed insignificantly in 24 h after infection and significantly decreased in 48 and 72 h under the action of H5N2 and H9N2, which correlated with manifestation of the cytopathogenic effect of these viruses. H5N2 and H9N2 avian viruses in THP-PMA macrophages are strong activators of the expression of the gene of the cytoplasmic RIG1 receptor 24 and 48 h after infection, and the pandemic virus H1N1pdm09 is a weak stimulator of RIG1 gene. Avian influenza H5N2 and H9N2 viruses are released by rapid induction of the inflammatory response in macrophages. At the late stages of infection, we observed a minor increase in IL-10 secretion in macrophages and, probably, the polarization of a part of the population in type M2. The studied influenza A viruses are weak inductors of IFN in THP-PMA macrophages. In the culture medium of THP-PMA macrophages infected with H9N2 and H5N2 viruses, MTT test revealed high levels of toxic factors causing the death of Caco-2 cells. In contrast to avian viruses, pandemic virus H1N1pdm09 did not induce production of toxic factors.


Assuntos
Vírus da Influenza A Subtipo H1N1/patogenicidade , Vírus da Influenza A Subtipo H5N2/patogenicidade , Vírus da Influenza A Subtipo H9N2/patogenicidade , Macrófagos/citologia , Macrófagos/metabolismo , Animais , Células CACO-2 , Diferenciação Celular/fisiologia , Humanos , Vírus da Influenza A Subtipo H1N1/imunologia , Monócitos/citologia , Monócitos/metabolismo , Pandemias
3.
Vopr Virusol ; 61(1): 21-6, 2016.
Artigo em Russo | MEDLINE | ID: mdl-27145596

RESUMO

The innate immune receptors TLR4, TLR7, TLR8, and RIG1 recognized the structures of the influenza viruses in human lymphocytes and were activated by the recombinant avian influenza virus A/Vietnam/1203/04 and its escape-mutant m13(13) during early period of interaction. The stimulated levels are not connected with viral reproduction. Donor cells with the low constitutive immune receptors gene expression levels showed higher stimulation. Inflammation virus effects resulted in. increasing production of TNF-alpha and IFN-gamma by lymphocytes. Signaling gene reactions of the parent and mutant viruses endosomal as well as cytoplasmic receptors are very similar. The mutant virus A/Vietnam/1203/04 (HA S145F), stimulated an increase in the transcription level of the membrane receptor gene TLR4 and a decrease in the level of activation of TNF-alpha gene. Further studies of natural influenza virus isolates are necessary to estimate the role of HA antigenic changes on immune reactions in humans.


Assuntos
Interações Hospedeiro-Patógeno/imunologia , Virus da Influenza A Subtipo H5N1/imunologia , Linfócitos/imunologia , Transdução de Sinais/imunologia , Regulação da Expressão Gênica , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Humanos , Imunidade Celular , Virus da Influenza A Subtipo H5N1/genética , Virus da Influenza A Subtipo H5N1/crescimento & desenvolvimento , Interferon gama/genética , Interferon gama/imunologia , Ativação Linfocitária , Linfócitos/virologia , Mutação , Cultura Primária de Células , Receptores do Ácido Retinoico/genética , Receptores do Ácido Retinoico/imunologia , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/imunologia , Receptor 7 Toll-Like/genética , Receptor 7 Toll-Like/imunologia , Receptor 8 Toll-Like/genética , Receptor 8 Toll-Like/imunologia , Transcrição Gênica , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia
4.
Eksp Klin Gastroenterol ; (9): 81-85, 2016.
Artigo em Inglês, Russo | MEDLINE | ID: mdl-29889402

RESUMO

The article presents data concerning the prevalence, clinical manifestations, methods of diagnostics, treatment and prevention of cholestasis of pregnancy, as well as the case management depending on the gestation and the cholestasis severity. A clinical case of a typical course of cholesrasis of pregnancy is presented.


Assuntos
Colestase , Complicações na Gravidez , Colestase/diagnóstico , Colestase/epidemiologia , Colestase/terapia , Feminino , Humanos , Gravidez , Complicações na Gravidez/diagnóstico , Complicações na Gravidez/epidemiologia , Complicações na Gravidez/terapia , Prevalência
5.
Eksp Klin Gastroenterol ; (9): 91-97, 2016.
Artigo em Inglês, Russo | MEDLINE | ID: mdl-29889404

RESUMO

The purpose of the review is to summarise the current data on chronic and acute pancreatitis with the goal to improve the diagnostics and treatment of women during pregnancy, which complicates differential diagnosis of hepatopancreatobiliary system pathology. The features of the incidence, etiology, parhogenesis and evaluation of the severity of the clinical manifestations of acute pancreatitis in pregnant women are given, It is emphasis that the most frequent its reason is the gallstones following by the biliary pancreatitis. The experience of use in pregnant patients imaging methods - endoscopic ultrasound, magnetic resonance cholangiopancrearography and endoscopic retrograde cholangiopancreatography, as well as treatments that include endoscopic sphincterotomy, stone extraction from the common bile duct and laparoscopic cholecystectomy. The recommendations are based on expert opinions and are not supported by randomized controlled trials. Nevertheless more active clinical management allows to diagnose and treat effectively pancreatitis, including biliary etiology, which contributes to a sharp decline in maternal and perinatal mortality.


Assuntos
Pancreatite/diagnóstico , Pancreatite/terapia , Complicações na Gravidez/diagnóstico , Complicações na Gravidez/terapia , Feminino , Humanos , Gravidez
6.
Artigo em Russo | MEDLINE | ID: mdl-25536769

RESUMO

AIM: Study the effect of inactivated influenza vaccines on the activity of innate and adaptive immunity genes (TLR3, TLR4 and B2M), RNA-interference Dicer1-gene, production of cytokines (antiviral IFN type I and II, regulatory IL10, IL17) and pro-inflammatory factors IL1-ß, TNFα. MATERIALS AND METHODS: Gene expression was determined by rRT-PCR with authors' primers in human blood cells treated with various doses of the vaccines. Concentration of cytokines by enzyme immunoassay was measured in cultural fluid using "Vector-best" kits. RESULTS: The studied vaccines have characteristic effects on genetic level. Grippol vaccine predominately stimulates TLR4 gene, activates TLR3, B2M and Dicer1 genes. Influvac vaccine mostly induces TLR3 gene and to a lesser extent TLR4 gene, does not influence the expression of B2M gene and inhibits Dicer1 gene. Vaxigrip split vaccine--the most potent stimulator of gene activity at low doses. Its main targets are TLR3 and B2M genes. All the inactivated vaccines--inductors of high level of IFNγ, low level of TNFα and do not induce IL17. Grippol additionally stimulates secretion of IL1-ß, and Vaxigrip - IFNα. Subunit vaccines Grippol and Influvac that contain purified influenza virus hemagglutinins induce IL10 synthesis in blood cells. CONCLUSION: Immunogenetic characteristics of the inactivated influenza vaccines administered nowadays are obtained.


Assuntos
Imunidade Adaptativa/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Imunidade Inata/genética , Influenza Humana/prevenção & controle , Vacinas/administração & dosagem , Imunidade Adaptativa/efeitos dos fármacos , Células Sanguíneas/efeitos dos fármacos , Citocinas/biossíntese , Humanos , Imunidade Inata/efeitos dos fármacos , Vírus da Influenza A/efeitos dos fármacos , Vírus da Influenza A/imunologia , Vírus da Influenza A/patogenicidade , Vacinas contra Influenza/administração & dosagem , Influenza Humana/imunologia , Influenza Humana/patologia , Interferons/biossíntese , Vacinas de Produtos Inativados/administração & dosagem
7.
Bull Exp Biol Med ; 156(2): 213-6, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24319751

RESUMO

The effects of Ridostin on the transcription of IFN family genes in human fibroblasts and lymphocytes were studied by quantitative real-time PCR. The degree of gene induction by Ridostin was most pronounced in fibroblasts, and was significantly higher than the induction by Kagocel: transcription of IFN-ß, oligoadenylate synthetase, and double-stranded RNA-dependent protein kinase genes increased by about 2000, 100, and 20 times, respectively. In lymphocytes, Ridostin also activated a wide variety of IFN family genes, including genes of IFN-ß, IFN-γ, and IFN-dependent enzymes, but this induction was less pronounced than in the fibroblasts. It was shown that gene response in lymphocyte from a child with cancer is reduced in comparison with that of adult healthy participant. Ridostin, and even more so Reaferon up-regulated activities of ß-actin, glycerophosphate dehydrogenase, and ß2-microglobulin genes, thus making impossible or limiting their use as constitutive stable reference genes (standards) in PCR-assays of IFN and their inductors.


Assuntos
Indutores de Interferon/farmacologia , Interferons/biossíntese , RNA de Cadeia Dupla/farmacologia , RNA Fúngico/farmacologia , Transcrição Gênica/efeitos dos fármacos , Ativação Transcricional/efeitos dos fármacos , 2',5'-Oligoadenilato Sintetase/biossíntese , 2',5'-Oligoadenilato Sintetase/genética , Actinas/biossíntese , Actinas/genética , Adulto , Antivirais/farmacologia , Linhagem Celular , Criança , Fibroblastos/metabolismo , Glicerolfosfato Desidrogenase/biossíntese , Glicerolfosfato Desidrogenase/genética , Gossipol/análogos & derivados , Gossipol/farmacologia , Humanos , Interferon alfa-2 , Interferon-alfa/farmacologia , Interferon beta/biossíntese , Interferon beta/genética , Interferon gama/biossíntese , Interferon gama/genética , Interferons/genética , Linfócitos/metabolismo , Vírus Elberfeld do Camundongo/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Microglobulina beta-2/biossíntese , Microglobulina beta-2/genética , eIF-2 Quinase/biossíntese , eIF-2 Quinase/genética
8.
Vopr Virusol ; 57(2): 27-31, 2012.
Artigo em Russo | MEDLINE | ID: mdl-22834144

RESUMO

The active replication of Karelian fever virus (KFV) in human blood vessels and the protective activity of the Russian agent reaferon were first shown. KFL was highly susceptible to interferon (IFN)-alpha. In control (uninfected) cells, reaferon caused low gene expressions of the IFN-dependent enzymes dsRNA-dependent protein kinase and 2'5'-oligoadenylate synthetase, by exerting a little effect on the activity of its family genes. KFV suppressed the reaferon-induced gene expression of IFN-dependent enzymes, but IFN-alpha gene transcription was increased in the reaferon-treated infected cells.


Assuntos
Células Sanguíneas/virologia , Interferon-alfa/farmacologia , Sindbis virus/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , 2',5'-Oligoadenilato Sintetase/metabolismo , Adolescente , Adulto , Infecções por Alphavirus/metabolismo , Animais , Células Sanguíneas/metabolismo , Chlorocebus aethiops , Feminino , Humanos , Interferon alfa-2 , Interferon-alfa/metabolismo , Masculino , Pessoa de Meia-Idade , RNA Viral/análise , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Sindbis virus/isolamento & purificação , Células Vero , eIF-2 Quinase/metabolismo
9.
Tsitologiia ; 52(2): 168-76, 2010.
Artigo em Russo | MEDLINE | ID: mdl-20352700

RESUMO

The aim of the study was to compare the dynamics ofmitochondrial apoptosis (MA) in cells at different stages of proliferation and with different susceptibility to cytomegalovirus (CMV). It has been found that in quiescent human fibroblasts (HF) CMV regulates MA at the level of bcl-2 gene transcription, exerting both pro- and anti-apoptotic effects. Suppression of bcl-2 transcription is greater in HF-977 line, which is highly susceptible to CMV in comparison with HF-1068 line. The effect of proliferative activity on MA was studied using CMV-infected HF-110044 line at the G0- or S-phase. A direct correlation was established between accumulation of cytochrome c and caspase 3 (MA markers) and production of IE72, pp65 and gB (CMV proteins). In G0-fibrob-lasts, viral replication was highly productive and bcl-2 expression was 10-fold as high as in S-phase cells, in which viral protein production and cell death were much lower. The increased gene transcription and accumulation of Bcl-2 protein enhanced cell viability and provided synthesis of viral proteins. Impaired structure of actin microfilaments, a caspase 3 target, coincided with pronounced suppression of gamma-actin gene in S-phase HF-110044. Our findings provide an insight into CMV-induced mechanisms of MA which lead to rapid death of infected quiescent fibroblasts and to slow death of cells infected at the stage of DNA synthesis.


Assuntos
Apoptose/genética , Infecções por Citomegalovirus/genética , Citomegalovirus , Mitocôndrias/fisiologia , Proteínas Proto-Oncogênicas c-bcl-2/genética , Actinas/genética , Actinas/metabolismo , Linhagem Celular , Infecções por Citomegalovirus/patologia , Fibroblastos/metabolismo , Fibroblastos/patologia , Fibroblastos/virologia , Regulação da Expressão Gênica , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transcrição Gênica
10.
Artigo em Russo | MEDLINE | ID: mdl-19004284

RESUMO

Cytomegalovirus (CMV) infection development and mRNA fas transcription levels (CD95) in resting (GO) and proliferating (S-phase) human lung embryo fibroblasts (HLEF-110044 line) were studied. In GO cells accumulation of infectious CMV was high and cell death was very quick, and fas gene expression was inhibited in early period of infection. In cells infected during S-phase CMV synthesis was lower and total cell death was detected only after 5 days; fas gene activity remained on high levels and increased during 6-48 hours. Death of CMV-infected fibroblasts occurred through apoptosis with cytopathic effect and detachment of cells in early stage, but without changes of cell membrane permeability and internucleosome fragmentation of DNA during later stages. In another HLEF-977 line CMV-induced apoptosis correlated with increased levels of fas gene transcription in resting cells. Positive association of activation Fas-receptor pathway and cell proliferation as well as different effect of CMV on activity of fas gene in 2 HLEF lines are discussed.


Assuntos
Apoptose , Infecções por Citomegalovirus/genética , Infecções por Citomegalovirus/patologia , Expressão Gênica , Receptor fas/genética , Linhagem Celular , Proliferação de Células , Citomegalovirus/fisiologia , Efeito Citopatogênico Viral , Fibroblastos/metabolismo , Fibroblastos/patologia , Fibroblastos/virologia , Humanos , Fase de Repouso do Ciclo Celular , Fase S , Fatores de Tempo , Replicação Viral
11.
Vopr Virusol ; 52(1): 28-36, 2007.
Artigo em Russo | MEDLINE | ID: mdl-17338231

RESUMO

Great differences were found in the level of 2',5'-oligoadenylate synthetase 40-46 kDa (OAS1) mRNA in relation to the proliferative state of human diploid fibroblasts at the moment of cytomegalovirus (CMV) (the strain AD169) infection. In the phase of synthesis of cellular cycle DNA (S), CMV induced OAS1 mRNA transcription by 10-100 times stronger than then in phase Go infection. The level of viral induction OAS1 mRNA peaked by hour 12 postinfection. The high gene activity correlated with suppressed DNA synthesis, a slowing-down mitotic cycle, markedly inhibited CMV replication, and delayed cell death. When the cells were infected in phase Go, the stimulation of OAS1 gene activity was less and it was attended by intensive viral replication and rapid cell death. There was a direct relationship between the resistance of cells and the constitutive level of OAS1 gene expression: in the low CMV-sensitive cells, the activity of OAS1 gene was more than 10 times greater than that in the highly sensitive cells. The inhibitors of the enzymatic OAS activity induced by IFN and dsRNA were found in the cytoplasm of the CMV-infected cells.


Assuntos
2',5'-Oligoadenilato Sintetase/genética , Infecções por Citomegalovirus/virologia , Citomegalovirus/fisiologia , Regulação da Expressão Gênica , 2',5'-Oligoadenilato Sintetase/química , 2',5'-Oligoadenilato Sintetase/metabolismo , Morte Celular , Linhagem Celular , Infecções por Citomegalovirus/metabolismo , Fibroblastos/citologia , Fibroblastos/metabolismo , Fibroblastos/patologia , Fibroblastos/virologia , Humanos , Interferon-alfa/metabolismo , Peso Molecular , RNA de Cadeia Dupla/metabolismo , RNA Mensageiro/genética , Fase de Repouso do Ciclo Celular/fisiologia , Fase S/fisiologia , Especificidade da Espécie , Replicação Viral/fisiologia
12.
Vopr Virusol ; 50(2): 32-6, 2005.
Artigo em Russo | MEDLINE | ID: mdl-15881395

RESUMO

The action of amixin and cycloferon on the expression of genes in the systems of interferon (IF) and cell apoptosis (CA) was studied by semi-quantitative RT-PCR in human blood microsamples before and after the administration of the drugs. Individual changes were determined in the transcription activity of genes of IF (alpha, beta, gamma), enzymes 2',5' oligoadenylatesynthetase (OAS), RNSase L, dsRNA-dependent proteinkinase (dsPK) and of CA effectors (FasAg, bcl-2, gamma-actin) registered dynamically in 24 h and 48 h. The activity parameters of IF genes were compared with the results of biological titration of IF activity in blood samples in vivo and in vitro. A pronounced ability of cycloferon to stimulate selectively the activity of genes of human IF, type I (beta IF--by 100 times and alpha IF--by 10 times), without affecting essentially the activity of other genes in blood cells, was detected. Amixin was found to inhibit the titration of genes with high activity levels. (alpha-, beta-IF, RNAases L, bcl-2 and gamma-actin). The antiviral and IF-induced properties of the drug are explained to a great extent by the apoptotic effect (activation of genes Fas, gamma-IF, OAS and affected transcription of gene bcl-2). A positive correlation was observed between the processes of activation of IF-genes transcription and the production of the total circulating IF. Antagonistic relations between type I and II IFs in human blood cells were shown.


Assuntos
Acridinas/farmacologia , Antivirais/farmacologia , Células Sanguíneas/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Indutores de Interferon/farmacologia , Interferon-alfa/genética , Interferon beta/genética , Interferon gama/genética , Tilorona/farmacologia , Acridinas/administração & dosagem , Administração Oral , Antivirais/administração & dosagem , Apoptose , Células Sanguíneas/metabolismo , Células Sanguíneas/fisiologia , Feminino , Humanos , Técnicas In Vitro , Injeções Intramusculares , Indutores de Interferon/administração & dosagem , Interferon-alfa/sangue , Interferon beta/sangue , Interferon gama/sangue , Pessoa de Meia-Idade , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tilorona/administração & dosagem , Transcrição Gênica/efeitos dos fármacos
13.
Vopr Virusol ; 50(1): 19-23, 2005.
Artigo em Russo | MEDLINE | ID: mdl-15747866

RESUMO

The interferon (IF) and cell-apoptosis (CA) systems are interrelated and regulate the protective reactions in body by means of a complex of biologically active proteins. The transcription levels of mRNA were determined by the RT-PCR semi-quantitative method in order to compare the constitutive expression levels of IF (alpha, beta, gamma) genes, IF-dependent enzymes of 2'5'-oligoadenylatesynthetyase (OAS), RNAase L, dsRNA-protein kinase (dsPK) and CA effectors (Fas-Ag, bcl-2 and gamma-actin) in human blood microsamples. cDNA dilutions, different numbers of amplification cycles (PCR with specific pairs of primers) as well as PCR-products' dilutions for dot-hybridization with specific probes were made use of to detect and evaluate levels of 9 mRNAs. The constitutive levels of gene expression of the IF and CA systems were found to differ essentially from others (1000-fold). The studied mRNA types were shared between 5 groups according to their transcription levels: very high--alpha-IF, high--RNAase L, medium--gamma-actin and bcl-2, low--beta-IF and very low (detectable after induction only)--gamma-IF, OAS, dsPK and Fas-Ag. The used detection method has a sufficiently high sensitivity and can be recommended for studies of IF inductors with unknown action mechanisms.


Assuntos
Apoptose/genética , Células Sanguíneas/fisiologia , Interferons/genética , 2',5'-Oligoadenilato Sintetase/biossíntese , 2',5'-Oligoadenilato Sintetase/genética , Actinas/biossíntese , Actinas/genética , Adulto , Doadores de Sangue , Primers do DNA , Endorribonucleases/biossíntese , Endorribonucleases/genética , Feminino , Expressão Gênica , Humanos , Interferons/biossíntese , Masculino , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , eIF-2 Quinase/biossíntese , eIF-2 Quinase/genética , Receptor fas/biossíntese , Receptor fas/genética
15.
Vopr Virusol ; 42(3): 98-102, 1997.
Artigo em Russo | MEDLINE | ID: mdl-9297349

RESUMO

Expression of 2,5-OAS and ds-protein kinase (ds-PK) in human fibroblast culture following treatment with alpha-IF in a dose of 1000 U or challenge with Karelian fever virus (KFV) (5 PFU/cell, 4 h.p.i.) was compared. A highly sensitive and rapid method, coupled RT-PCR, was used to detect mRNA transcription levels. Alpha-IF had an expressed stimulating effect on 2,5-OAS mRNA level, whereas the virus markedly inhibited the synthesis of ds-PK mRNA.


Assuntos
2',5'-Oligoadenilato Sintetase/genética , Regulação Enzimológica da Expressão Gênica , Interferon-alfa/farmacologia , Proteínas Serina-Treonina Quinases/genética , Sindbis virus/fisiologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/enzimologia , Fibroblastos/virologia , Humanos , Reação em Cadeia da Polimerase , RNA Mensageiro/genética , Transcrição Gênica , eIF-2 Quinase
17.
Vopr Virusol ; 41(6): 245-52, 1996.
Artigo em Russo | MEDLINE | ID: mdl-8999680

RESUMO

A recombination of the genomes of VEE and KF viruses, two alphaviruses of different serotypes, takes place in mixed infection of chick embryo fibroblasts and Vero cells with vaccine strain VEE-230 and KF-9298 strain. This recombination resulted in the production of recombinant virions with altered antigenic properties. Polymerase chain reaction (PCR) rapidly and effectively detected the hybrid sites of the genome in recombinant viruses. Short nucleotide sequences (approximately 20 n. p.) of a known primary structure in the RNA genomes, specifically reacting with the complementary PCR primers, were chosen as genetic markers. These data are the first experimental validation of the probable recombinant formation of alphaviruses in nature. They are in good agreement with findings of the recombinant structure of Western equine encephalomyelitis virus genome.


Assuntos
Alphavirus/genética , Vírus da Encefalite Equina Venezuelana/genética , Nucleocapsídeo/genética , RNA Viral/genética , Recombinação Genética , Proteínas do Envelope Viral/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Embrião de Galinha , Chlorocebus aethiops , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Células Vero
18.
Vopr Virusol ; 41(5): 209-14, 1996.
Artigo em Russo | MEDLINE | ID: mdl-8967065

RESUMO

Genetic relationships of geographical isolates of the members of WEE virus serocomplex (McMillan, Fort Morgan, Highlands J, and Y62-33) were assessed by the polymerase chain reaction (PCR) and restriction analysis of the PCR products. Oligonucleotide primers (21 nucleotides in length) were chosen for NSP2, nucleocapsid C, and E2-E1 protein genes based on the known primary structure of the McMillan 16310-5614 genome (L. Uryvayev et al., 1994, 1995). These primers were shown to differentiate well the WEE and SV-like strains of the serocomplex. Y62-33 virus (Udmurtia, Russia) was identical to McMillan strain in three studied regions of NSP2, C, and E2-E1 genes. NSP2 gene could be detected in all the studied geographical isolates and was characterized by the same restriction patterns as endonucleases; it appeared to be the most conservative. The structural genes were less conservative. Fort Morgan virus (Colorado, USA) genome reliably differed from McMillan virus (California, USA) and was negative in PCR with primers to C and E2 gene regions. Highlands J genome (Florida, USA) was positive in PCR with the primers to E2-E1 gene regions but differed from McMillan strain by the nucleocapsid gene. An additional comparative PCR analysis of the C-E2 region in the McMillan and Highlands J genomes showed some, but not complete identity. The origin of these two viruses might be due to the selection of different forms of recombinant viruses. A good correlation of structural genes in PCR and the infectivity neutralization test was noted with the primers and polyclonal antibodies to the closely related strains. High specificity of PCR permits a more accurate detection of the virus origin and relationships.


Assuntos
Vírus da Encefalite Equina do Oeste/genética , Genes Virais , Proteínas do Core Viral/genética , Proteínas do Envelope Viral/genética , Proteínas não Estruturais Virais/genética , Proteínas Estruturais Virais/genética , Animais , Linhagem Celular , Vírus da Encefalite Equina do Oeste/classificação , Variação Genética , Sondas de Oligonucleotídeos , Reação em Cadeia da Polimerase , Sorotipagem
19.
Vopr Virusol ; 41(3): 117-22, 1996.
Artigo em Russo | MEDLINE | ID: mdl-8928504

RESUMO

Comparison of Sindbis virus strains isolated in different regions of the world (in Africa, Australia, and Europe, including Russia and its nearest neighbors) in the polymerase chain reaction (PCR) by the primary gene structure of proteins NSP1 and E1 and in the neutralization test showed the greatest similarity between geographically close strains isolated in Northern Europe (KFL, Karelia, 1381 and 1388, Estonia). Sindbis strains AR339 and Babanki isolated in Africa were similar to each other and to strains from Northern Europe by the examined gene sites but different from the Northern variants in the neutralization test. Geographically remote strains F-720 (Armenia and Southern Europe) and Whataroa (New Zealand) were close to Sindbis virus from Africa and Northern Europe by only one of the genes examined (F-720 by NSP1 and Whataroa by E1). PCR was carried out using oligonucleotide primers containing nucleotide sequences identical to genes NSP1 and E1 sites of Sindbis strains HRSP, Okelbo, and KFL, but different from gene sites of other known representatives of alphaviruses by at least 5 positions. PCR analysis showed that the appurtenance of the geographic variants to Sindbis group can be ascertained only after investigating the homology of at least two genes coding for the replicative and structural proteins. Such a procedure of PCR permits the detection of Sindbis viruses of different geographic origin with changes in their primary structure and allows the differentiation between Sindbis viruses and Western equine encephalomyelitis viruses within the serological complex.


Assuntos
Vírus da Encefalite Equina do Oeste/genética , Sindbis virus/genética , Vírus da Encefalite Equina do Oeste/isolamento & purificação , Glicoproteínas de Membrana/genética , Reação em Cadeia da Polimerase , RNA Viral/genética , Sindbis virus/isolamento & purificação , Especificidade da Espécie , Proteínas do Envelope Viral/genética , Proteínas não Estruturais Virais/genética
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