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1.
A repressor of interferon production is contained in the hyaloplasm of the chick embryo fibroblast cells in the state of hyporeactivity. A factor stimulating interferon production (FSIP) was found in the hyaloplasm of the cells subjected to single or double induction with poly (pI) x poly (pC) followed by exposure to actinomycin D. Both preparations were of the protein nature and possessed high biological activity. They repressed or stimulatd interferon production at a dilution of 1:512-1:1024. The preparations had tissue and inductor specificity, did not affect the Venezuelan equine encephalomyelitis virus and interferon antiviral activity. Unlike the repressor, the FSIP proved to be thermolabile and pH-sensitive in acid media. The half lives of the repressor and stimulator were about 10 and 5 hours respectively. 相似文献
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Novel original preparations of double-stranded RNAs (dsRNAs), i.e. larifan, ridostin and rifastin, and recombinant alpha 2- and beta-interferons promising for the clinical use were studied. The size and morphology of the dsRNAs in the preparation composition, the dynamics of their induction of interferon and the antiviral state in human fibroblasts and the effect of the DEAE dextran polycation on the activity of the dsRNAs were specified. For the first time the dynamics of 2',5'-oligoadenylate synthetase activity in the nuclei and cytoplasm of the human fibroblasts treated with the dsRNAs of different origin and their complexes with DEAE dextran was defined. To elucidate the specific features of the mechanism of antiviral action of dsRNAs and interferon, the relation of the 2',5'-oligoadenylate synthetase activity to dsRNAs was investigated. In the cells treated with dsRNAs and DEAE dextran there were an early activation of the enzyme and predominance of the enzyme activated forms requiring no addition of poly I.poly C to the reaction mixture. The results were indicative of possible intracellular activation of its isoforms, similar to that in the cells treated with interferon and contaminated with viruses. All the tested preparations of dsRNAs and interferons induced an increase in the activity of 2',5'-oligoadenylate synthetase both in the cytoplasm and the nuclei of human fibroblasts. The same ability was observed in DEAE dextran which is likely to be one of the causes of the increase in dsRNAs antiviral activity under its effect. 相似文献
3.
T M Sokolova M G Gevorkian E B Tazulakhova F I Ershov 《Molekuliarnaia genetika, mikrobiologiia i virusologiia》1985,(10):43-46
The hyporeactivity factor in interferon production by L-929 cells designated IRP (interferon repressing protein) has been studied. In particular, its localization and methods of its purification have been studied. The kinetics of IRP accumulation by producing cells correlate with the development of hyporeactivity condition. Most of IRP is localized in cell sap and in ribosomal fraction in evidence to regulatory role of repressor at the level of interferon mRNA translation. A 100-fold increase in repressor activity was achieved by IRP concentration by ammonium sulfate precipitation. IRP as well as interferon have been shown to possess high affinity to polyU sepharose. The preparations of IRP and interferon concentrated by ammonium sulfate precipitation were subsequently purified by fractioning in a polyI sepharose column. A 10,000-fold (6 x 10(4) U/mg) purification was achieved for IRP and 250-fold (10(4) U/mg) for interferon. 相似文献
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T G Orlova T V Mamontova L V Zhdanova V D Solov'ev 《Biulleten' eksperimental'no? biologii i meditsiny》1977,83(4):436-437
Chick embryo cultures treated with interferon yielded a biologically active RNA which, upon inoculation into chick embryo cells, created an antiviral condition in them. The level of vesicular stomatitis virus reproduction in such cells was 2-30% of that observed in the cells treated with control RNA. The maximum activity of the experimental RNA was seen 3 hours after the treatment with interferon. 相似文献
6.
RNase L Mediates the Antiviral Effect of Interferon through a Selective Reduction in Viral RNA during Encephalomyocarditis Virus Infection 总被引:8,自引:4,他引:4
The 2′,5′-oligoadenylate (2-5A) system is an RNA degradation pathway which plays an important role in the antipicornavirus effects of interferon (IFN). RNase L, the terminal component of the 2-5A system, is thought to mediate this antiviral activity through the degradation of viral RNA; however, the capacity of RNase L to selectively target viral RNA has not been carefully examined in intact cells. Therefore, the mechanism of RNase L-mediated antiviral activity was investigated following encephalomyocarditis virus (EMCV) infection of cell lines in which expression of transfected RNase L was induced or endogenous RNase L activity was inhibited. RNase L induction markedly enhanced the anti-EMCV activity of IFN via a reduction in EMCV RNA. Inhibition of endogenous RNase L activity inhibited this reduction in viral RNA. RNase L had no effect on IFN-mediated protection from vesicular stomatitis virus. RNase L induction reduced the rate of EMCV RNA synthesis, suggesting that RNase L may target viral RNAs involved in replication early in the virus life cycle. The RNase L-mediated reduction in viral RNA occurred in the absence of detectable effects on specific cellular mRNAs and without any global alteration in the cellular RNA profile. Extensive rRNA cleavage, indicative of high levels of 2-5A, was not observed in RNase L-induced, EMCV-infected cells; however, transfection of 2-5A into cells resulted in widespread degradation of cellular RNAs. These findings provide the first demonstration of the selective capacity of RNase L in intact cells and link this selective activity to cellular levels of 2-5A. 相似文献
7.
I Táborsky J Doskocil D Stancek 《Journal of hygiene, epidemiology, microbiology, and immunology》1977,21(3):261-270
The dynamics of interferon formation by an established cell line of mouse fibroblast (L cells) and by mouse peritoneal leukocytes induced by double-stranded RNA extracted from E. coli f2 phage is described. The L cells produced interferon at a lower rate, the maximum values were obtained at 12 to 20 hours after induction, and the production was ultimately dependent on the established cell line used and on the presence of DEAE-dextran during induction. The mouse peritoneal leukocytes (MPL), on the other hand, did not require DEAE-dextran and the maximum of interferon production was reached between 6 and 12 hours after induction. Both the L cell- and the MPL-interferons were purified and concentrated so that the final specific biologic activity was 100-to 300-fold higher than that of the initial preparations (1 to 5 X 10(6) interferon units per mg protein). Polyacrylamide gel electrophoresis showed similar migration profiles for the preparations of both interferons. The smaller part of the activity was situated in a broader, slow-moving peak and the greater part formed a sharp, high and fast-moving peak. Using 3H uridine-labelled f2 ds-RNA for induction of interferon it was found that one of the radioactivity zones coincided with the fast-moving activity peak of the purified and concentrated interferon. 相似文献
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beta-Interferon alters the pattern of proteins secreted from quiescent and platelet-derived growth factor-treated BALB/c-3T3 cells 总被引:4,自引:0,他引:4
Mouse beta-interferon (at a concentration of 100 units/ml or higher) inhibited the platelet-derived growth factor (PDGF)-induced replication of quiescent BALB/c-3T3 cells. The interferon treatment did not inhibit, but slightly enhanced, the accumulation of the following three PDGF-induced RNAs: myc RNA, JE RNA, and KC RNA. The treatment with interferon changed the pattern of secreted proteins from quiescent cells and from cells treated with partially purified PDGF; it inhibited the accumulation of the PDGF-induced proteins (including proteins of 63 and 32 kDa) and it induced the accumulation of several other proteins (including proteins of 89, 31.5, 30, and 10.5 kDa) in both quiescent and also in PDGF-treated cells. 相似文献
10.
Abstract Five different interferon preparations were compared for their antiviral activity against Herpes simplex virus type 1 (HSV-1) and several RNA viruses. The interferons used were: interferon α from human buffy coats, interferon β from human fibroblasts, interferon γ from human lymphocytes after stimulation with phytohemagglutinin (PHA), lymphoblastoid interferon from Namalva cells IFN-α (Ly) and cloned α 2 interferon produced by Escherichia coli containing the human gene for interferon α 2 . All preparations were able to protect monolayers of HeLa cells against HSV-1 infection when low multiplicities were used. The five IFN preparations were also tested against encephalomyocarditis (EMC) virus, poliovirus and vesicular stomatitis virus (VSV). 相似文献
11.
The size of genomic RNA in PR-RSV A passaged in chick embryo fibroblasts (CEF) or quail embryo fibroblasts (QEF) was determined by gel electrophoresis. The results showed that 3 undiluted passages resulted in heterogeneity of RNA. The heterogeneity of the smaller incomplete RNAs in the virus stock was decreased by diluted passage or cloning, but RNA of the b subunit size and a subunit RNA of complete genome size were relatively stable. These heterogeneous RNAs were characterized by hybridization analysis. The RNAs from 4 peaks hybridized with both cDNAtotal and cDNAsrc to appreciable extents, indicating that they were derived from viral RNA and that at least some of them contained the src sequence. This finding and the failure to isolate a td mutant from the undiluted-passaged virus stock or from some subclones that had a and b subunits of RNA indicate that the td virus was only a minor constituent of the incomplete virus population caused by undiluted passages. Some viruses with incomplete RNA in the virus stock could produce foci with the aid of td B77 or RAV-60. The emergence of rd viruses by undiluted passages was indicated. 相似文献
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Krishnamurthy Malathi Takeshi Saito Nannette Crochet David J. Barton Michael Gale Jr Robert H. Silverman 《RNA (New York, N.Y.)》2010,16(11):2108-2119
Triggering and propagating an intracellular innate immune response is essential for control of viral infections. RNase L is a host endoribonuclease and a pivotal component of innate immunity that cleaves viral and cellular RNA within single-stranded loops releasing small structured RNAs with 5′-hydroxyl (5′-OH) and 3′-monophosphoryl (3′-p) groups. In 2007, we reported that RNase L cleaves self RNA to produce small RNAs that function as pathogen-associated molecular patterns (PAMPs). However, the precise sequence and structure of PAMP RNAs produced by RNase L is unknown. Here we used hepatitis C virus RNA as substrate to characterize RNase L mediated cleavage products [named suppressor of virus RNA (svRNA)] for their ability to activate RIG-I like receptors (RLR). The NS5B region of HCV RNA was cleaved by RNase L to release an svRNA that bound to RIG-I, displacing its repressor domain and stimulating its ATPase activity while signaling to the IFN-β gene in intact cells. All three of these RIG-I functions were dependent on the presence in svRNA of the 3′-p. Furthermore, svRNA suppressed HCV replication in vitro through a mechanism involving IFN production and triggered a RIG-I-dependent hepatic innate immune response in mice. RNase L and OAS (required for its activation) were both expressed in hepatocytes from HCV-infected patients, raising the possibility that the OAS/RNase L pathway might suppress HCV replication in vivo. It is proposed that RNase L mediated cleavage of HCV RNA generates svRNA that activates RIG-I, thus propagating innate immune signaling to the IFN-β gene. 相似文献
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Defective interfering influenza viruses and host cells: establishment and maintenance of persistent influenza virus infection in MDBK and HeLa cells. 总被引:7,自引:5,他引:2
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WSN (H0N1) influenza virus upon undiluted passages in different species of cells, namely, bovine kidney (MDBK), chicken embryo (CEF), and HeLa cells, produced a varying amount of defective interfering (DI) virus which correlated well with the ability of the species of cell to produce infectious virus. However, the nature of the influenza DI viral RNA produced from a single clonal stock was essentially identical in all three cells types, suggesting that these cells do not exert a great selective pressure in the amplification of specific DI viral RNAs either at early or late passages. DI viruses produced from one subtype (H0N1) could interfere with the replication of infectious viruses belonging to other subtypes (H1N1, H3N2). DI viral RNAs could also replicate with the helper function of other subtype viruses. The persistent infection of MDBK and HeLa cells could be initiated by coinfecting cells with both temperature-sensitive mutants (ts-) and DI influenza viruses. Persistently infected cultures cultures at early passages (up to passage 7) showed a cyclical pattern of cell lysis and virus production (crisis), whereas, at later passages (after passage 20), they produced little or no virus and were resistant to infection by homologous virus but not by heterologous virus. The majority of persistently infected cells, however, contained the complete viral genome since they expressed viral antigens and produced infectious centers. Selection of a slow-growing temperature-sensitive variant rather than the presence of DI virus or interferon appears to be critical in maintaining persistent influenza infection in these cells. 相似文献
15.
The effects of Escherichia coli RNA polymerase and its subassemblies and subunits on the in vitro synthesis of beta subunit directed by DNA from a lambda transducing phage lambdadrif+-6 were investigated. This phage carries the structural gene (rpoB) for beta subunit as well as the genes for EF (translation elongation factor)-Tu, some ribosomal proteins, and stable RNAs of the E. coli chromosome. Among the RNA polymerase proteins examined, the two oligomers, holoenzyme and alpha2beta complex, repressed the synthesis of only the beta subunit but not of other proteins encoded by the phage DNA. The results indicate that the expression of at least the betabeta' (rpoBC) operon is under autogenous regulation, in which both holoenzyme and alpha2beta complex function as regulatory molecules with repressor activity. 相似文献
16.
The Miyadera strain of Newcastle disease virus (NDV) consisted predominantly of virus particles forming small plaques on monolayers of chick embryo fibroblasts (CEF), and contained small amounts of virus particles forming large plaques. These large- and small-plaque-forming clones of this virus (NDV-L and NDV-S) were isolated. The small size of the NDV-S plaques did not appear to be due to an agar inhibitor. NDV-L produced a much higher yield of infective virus particles in CEF and they were released more completely from the infected cells than were those produced by NDV-S. The yield of infective virus of NDV-L per cell from cultures of CEF was comparable to the yield from the allantoic cells. The infectivity/hemagglutinin ratio for NDV-L from CEF was as high as the ratio for virus from the allantoic cells, but the ratio for NDV-S from CEF was lower. NDV-S demonstrated an autointerference phenomenon in CEF when infected at high multiplicities, but NDV-L did not. Contrary to virus multiplication, NDV-S exhibited a more rapid and marked cytopathic effect on monolayers of CEF than NDV-L. In the allantoic cavity of eggs NDV-S produced slightly higher virus yields than NDV-L. No correlation existed between plaque size of the two viruses and the capacity to induce interferon synthesis or the susceptibility to the action of interferon. The properties of both distinctive plaque isolates were stable on egg passage. 相似文献
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Evidence of Avian Leukosis Virus Subgroup E and Endogenous Avian Virus in Measles and Mumps Vaccines Derived from Chicken Cells: Investigation of Transmission to Vaccine Recipients 总被引:2,自引:0,他引:2
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Shirley X. Tsang William M. Switzer Vedapuri Shanmugam Jeffrey A. Johnson Cynthia Goldsmith Anthony Wright Aly Fadly Donald Thea Harold Jaffe Thomas M. Folks Walid Heneine 《Journal of virology》1999,73(7):5843-5851
20.
Heterogeneity of interferon mRNA species from Sendai virus-induced human lymphoblastoid (Namalva) cells and Newcastle disease virus-induced murine fibroblastoid (L) cells. 总被引:4,自引:0,他引:4
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A D Sagar L A Pickering P Sussman-Berger W E Stewart nd P B Sehgal 《Nucleic acids research》1981,9(1):149-160
Cytoplasmic polyadenylated RNA preparations obtained from Sendai-induced human lymphoblastoid (Namalva) cells and from Newcastle disease virus (NDV)-induced murine (L) cells were denatured in 10-12.5 mM CH3HgOH and then electrophoresed in 2% agarose tube gels containing 10 mM CH3HgOH, the RNA eluted from gel slices and translationally active interferon mRNA species located using the Xenopus oocyte assay. The interferons synthesized were characterized as alpha or beta types based on neutralization tests using specific antisera against human or murine interferon-alpha and interferon-beta. At least two species of mRNA for human interferon-alpha and two for human interferon-beta were detected in RNA from Sendai-induced Namalva cells. These are (approximate mRNA length in parentheses) alpha (1.3 kb), alpha (1.9 kb), beta (1.1 kb) and beta (1.9 kb). Two populations of murine interferon mRNA of lengths approximately 1.4 kb and 3 kb were detected in mRNA preparations from NDV-induced L cells by electrophoresis. However, since the translation products of each of these two populations of mRNA consist of both murine interferon-alpha and murine interferon-beta it is likely that both the 1.4 kb and 3 kb populations contain at least one species each of murine interferon-alpha and murine interferon-beta mRNA. 相似文献