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1.
2.
Impaired antiviral response in human hepatoma cells.   总被引:8,自引:0,他引:8  
Hepatitis B, C, and D viruses can infect liver cells and in some individuals establish a chronic phase of infection. Presently, relatively little information is available on the antiviral mechanisms in liver cells. Because no good in vitro model infection systems for hepatitis viruses are available, we have used influenza A, Sendai, and vesicular stomatitis (VSV) viruses to characterize interferon (IFN) responses and IFN-induced antiviral mechanisms in human hepatoma cell lines. HepG2 or HuH7 cells did not show any detectable IFN-alpha/beta production in response to influenza A or Sendai virus infections. Treatment of cells with IFN-alpha resulted in upregulation of IFN-alpha-inducible Mx, 2',5'-oligoadenylate synthetase (OAS) and HLA class I gene expression but only with exceptionally high levels of IFN-alpha (>/=100 IU/ml). Accordingly, high pretreatment levels of IFN-alpha, 1000 IU/ml for influenza A and VSV and 100 IU/ml for Sendai virus, were required before any detectable antiviral activity against these viruses was seen. IFN-gamma had some antiviral effect against influenza A virus but appeared to be ineffective against VSV and Sendai virus. IFN-gamma upregulated HLA class I protein expression, whereas Mx or OAS expression levels were not increased. There was a modest upregulation of HLA class I expression during Sendai virus infection, whereas influenza A virus infection resulted, after an initial weak upregulation, in a clear decrease in HLA class I expression at late times of infection. The results suggest that hepatoma cells may have intrinsically poor ability to produce and respond to type I IFNs, which may contribute to their inability to efficiently resist viral infections.  相似文献   

3.
We sought to investigate the anti-severe acute respiratory syndrome (SARS)-associated coronavirus (SCoV) activities of type I (alpha and beta) and type II (gamma) interferons (IFN) in vitro. Type I IFNs protected cells from cytopathic effects (CPE) induced by SCoV, and inhibited viral genomic RNA replication in FRhk-4 cells (measured by quantitative RT-PCR) in a dose-dependent manner. Intracellular viral RNA copies were reduced 50% by IFN-alpha at a concentration of 25 U/ml and by IFN-beta at a concentration of 14 U/ml. IFN-gamma had fewer effects on inhibition of viral infection and replication. The type I IFN receptor signaling pathway in host cells is mainly involved in the inhibition of SCoV infection and replication. Type I IFNs could be used as potential agents for anti-SARS treatment.  相似文献   

4.
The antiviral activity of three types of spotted souslik interferons So IFN alpha, So IFN beta and So IFN gamma was studied. Fibroblasts of lungs (SL), kidneys (SK) of adult sousliks and skin fibroblasts of foetuses (SE) were equally sensitive to three types of souslik interferons. In contrast to So IFN beta and So IFN gamma, So IFN alpha, exhibited a high (100% of activity in homologous cells) cross species antiviral activity in mouse embryo fibroblasts (MEF) and a low (3% to 10% of the activity) in bovine embryo fibroblasts (BEF) and human embryo fibroblasts (HEF). The development kinetics of antiviral activity not only depended on the interferon type but also on the temperature of incubation. In comparison with So IFN gamma, So IFN alpha and So IFN beta activated earlier the maximal antiviral state. Low incubation temperature (26 degrees C) did not decrease but only delayed the antiviral activity of spotted souslik interferons. Mixed preparations of So IFN gamma with So IFN alpha or So IFN beta exhibited synergistic antiviral activity at physiological (37 degrees C) and low (26 degrees C) temperatures. The development of antiviral activity of So IFN beta as well that of Mu IFN alpha/beta was inhibited by plant lectins which reacted with the cell membrane compounds. All three types of souslik interferons were completely destroyed by trypsin and boiling at 100 degrees C for 1 min. and partially by SDS. Their sensitivity to shaking, beta-mercaptoethanol and mouse antisera against So IFN beta was different in relation to the interferon type.  相似文献   

5.
Oral administration of type I interferons (IFNs; murine IFN-alpha and IFN-beta) reduces early replication of murine cytomegalovirus (MCMV) in both the spleen and liver of MCMV-infected BALB/c mice. Examination of a range of doses of IFN (1 to 1000 IU) showed that 10 IU administered daily for 1 week prior to virus infection was optimal for inhibition of MCMV replication. Furthermore, low-dose orally administered IFN (10 IU/day) was effective in mice challenged with lethal and sublethal virus inocula. The antiviral efficacy of low-dose orally administered IFN was not restricted by either the route of virus inoculation or the mouse genotype. Analysis by immunohistochemistry of IFN-alpha receptor-bearing cells of the gastrointestinal tract revealed predominant staining of perivascular smooth muscle and the lamina propria of the anterior tongue, small intestine and rectum. These tissues, dense in IFN-alpha receptor-bearing cells, are likely to be the sites of interaction of the orally administered IFNs with the mucosal immune system. In conclusion, we propose that low-dose oral use of type I IFN therapy may have broad applications in the treatment of CMV infections.  相似文献   

6.
M S Diamond  E Harris 《Virology》2001,289(2):297-311
Previously, we demonstrated that pretreatment of cells with interferon (IFN) alpha + gamma or beta + gamma inhibited dengue virus (DV) replication. In this study, experiments were performed to better define the mechanism by which IFN blocks the accumulation of dengue virus (DV) RNA. Pretreatment of human hepatoma cells with IFN beta + gamma did not significantly alter virus attachment, viral entry, or nucleocapsid penetration into the cytoplasm. The inhibitory effect of IFN was retained even when naked DV RNA was transfected directly into cells, confirming that steps associated with viral entry were not the primary target of IFN action. Biosynthetic labeling experiments revealed that IFN abolished the translation of infectious viral RNA that occurred prior to RNA replication. Subcellular fractionation experiments demonstrated that IFN did not significantly alter the ability of viral RNA to attach to ribosomes. The antiviral effect of IFN appeared independent of the IFN-induced, double-stranded RNA-activated protein kinase (PKR) and RNase L, as genetically deficient PKR- RNase L- cells that were infected by DV retained sensitivity to inhibition by IFN. We conclude that IFN prevents DV infection by inhibiting translation of the infectious viral RNA through a novel, PKR-independent mechanism.  相似文献   

7.
Replication of the diabetogenic variant of encephalomyocarditis virus (EMCV-D) in spleen cells and its association with subpopulations of spleen cells (L3T4+, Lyt-2+, Mac 1+, 33D1+ and AGM1+ cells) from both sexes of ICR Swiss mice were examined. Virus replication was limited to less than 0.5 log in suspensions of whole spleen cells, nonadherent cells or a B cell subfraction from both sexes of ICR Swiss mice following infection with EMCV-D at an MOI of 10; no virus replication was seen in adherent spleen cells from either sex. After 1 hour adsorption of EMCV-D onto spleen cells at a multiplicity of infection (MOI) of either 10 or 0.1, virus-associated cells were isolated using a monoclonal murine anti-EMCV-D and anti-mouse IgG conjugated to magnetic beads. Using an MOI of 0.1, less than 1% of spleen cells bound virus particles after 1 hour adsorption at 4 degrees C. Among the virus-positive cells, relatively higher percentages of adherent cell populations (Mac 1+ and 33D1+ cells) of both sexes bound virus particles within the first hour post-infection (PI) than did the other spleen cell subpopulations. Interferon (IFN) alpha/beta production was detected as early as 4 hours PI in female spleen cell cultures infected with EMCV-D at an MOI of 0.1 while no IFN alpha/beta activity was found in comparably infected male spleen cell cultures. Inhibiting IFN alpha/beta activity in the virus-infected spleen cell cultures during the first 20 hours of infection using polyclonal rabbit anti-mouse IFN alpha/beta serum eliminated production of IFN gamma as well as IFN alpha/beta. Spleen cell cultures depleted of adherent cells were unable to produce IFN alpha/beta or IFN gamma in the first 24 hours PI. The capacity to produce IFN gamma at 12 hours after virus infection of spleen cells from both sexes of mice was restored to adherent cell-depleted cultures by addition of mouse IFN alpha/beta at the time of infection. These results suggest that IFN alpha/beta and adherent cells play critical roles in the early production of IFN gamma (less than 16 hours PI) characteristic of the infected spleen cell cultures of females. Production of IFN alpha/beta and IFN gamma by spleen cells from both sexes of ICR Swiss mice was enhanced by administrating estrone to donor mice during the week before harvesting spleen cells.(ABSTRACT TRUNCATED AT 400 WORDS)  相似文献   

8.
It was found that development of antiviral state in cells activated by IFN alpha and beta were quite rapid. After 6-7 h treatment IFN alpha and IFN beta protected cells completely and their effects lasted for another 9-12 h after removing them from the cultures. Then the effect declined, but some protective action remained after 48 h of incubation. IFN-gamma was different from IFN alpha or beta. It activated cells much more slowly, and could not protect cells completely before 24 h of treatment. When we compared three concentrations of IFN-alpha, the cells could not be protected completely by 5 IU/ml, and kinetics were similar with 250 and 25 IU/ml. When we added anti-IFN-alpha serum before IFN treatment, the development of the antiviral state was inhibited. The results suggested that one should adopt different ways when titrating and using different forms of IFN, and it might not be needed to use maximal tolerated doses and daily administrations, and to keep the high level of IFN for a long time. We think that these results might be useful for the clinicians considering the optimal schedule for IFN treatment.  相似文献   

9.
Summary Genetic heterogeneous mouse populations selected for high (HIII) and low (LIII) antibody response were used to study some aspects of mouse hepatitis virus 3 (MHV3) infection, such as the resistance pattern, virus replication in the liver and peritoneal exudate or in cultured peritoneal macrophages, the interferon (IFN) synthesis in the serum and peritoneal exudate and the procoagulant activity (PCA) of the peritoneal exudate (PEC) and spleen cells (SC). The HIII mice, when compared to their LIII mice counterparts, were susceptible to MHV3 infection showing higher virus titres in the liver and peritoneal exudate, comparable IFN alpha/beta or IFN gamma titres in the peritoneal exudate or in the serum, and higher levels of PCA of PEC and SC. A higher virus titre was detected in the supernatants of HIII mouse macrophages infected with MHV3. The activation of HIII mouse macrophages with LPS, IFN alpha/beta or IFN gamma, in contrast to that of LIII mouse macrophages, did not induce an antiviral effect with partial restriction of the MHV3 replication. The LPS antiviral activity was shown to be partially exerted by IFN alpha/beta synthesis. The IFN gamma was shown to be more effective in inducing an antiviral state in LIII macrophages, when compared to IFN alpha/beta. The data obtained are consistent with the notion that the resistance mechanisms to the MHV3 infection involve the PCA and the sensitivity of macrophages to IFN.  相似文献   

10.
Viruses and the type I interferon antiviral system: induction and evasion   总被引:1,自引:0,他引:1  
The type I interferon (IFN) system responds to viral infection and induces an "antiviral state" in cells, providing an important first line of defense against virus infection. Interaction of type I IFNs (IFN alpha and IFN beta) with their receptor induces hundreds of cellular genes. Of the proteins induced by IFN, the antiviral function of only a few is known, and their mechanisms of action are only partly understood. Additionally, although viral-encoded mechanisms that counteract specific components of the type I IFN response have been known for some time, it has recently become clear that many (if not most) viruses encode some form of IFN-antagonist. Understanding the interplay between viral-encoded IFN antagonists and the interferon response will be essential if the therapeutic potential of IFNs is to be fully exploited.  相似文献   

11.
12.
Human lymphocytes pretreated with interferon (IFN) alpha, beta, or gamma produced 17 times more IFN alpha (600-10,000 units/ml) than nontreated lymphocytes when cocultivated with allogeneic cells. Significant increases in IFN production (500-3,000 units/ml) were observed when lymphocytes were treated with IFN for just 2 h, and peak levels (10,000 units/ml) were produced after a 4-h treatment. The amount of IFN required to show the maximum priming effect was between 100 and 1,000 units; higher levels of IFN were inhibitory. The levels of IFN increased as the lymphocyte-to-target-cell ratio increased from 2:1 to 10:1 and decreased at higher ratios. The decrease in IFN production at higher ratios of lymphocytes to target cells could not be attributed to the presence of a soluble suppressor substance. The additional IFN found in supernates was attributed to enhanced production of IFN by the same cells, rather than recruitment of more cells to produce IFN. This conclusion is based on the fact that no increase in the number of cells staining positive for IFN production was observed in primed lymphocytes. The increased amount of IFN due to priming enhanced both nonsensitized cytotoxic activity and the transfer of antiviral activity, which could be prevented by antibody to IFN. The data suggest that priming may be an important biological mechanism for obtaining significant levels of IFN more rapidly in the vicinity of transformed cells or virus-infected tissues.  相似文献   

13.
One mechanism by which interferons (IFNs) can inhibit chlamydial infection is by the induction of the enzyme indoleamine 2,3-dioxygenase (IDO), which restricts the availability of tryptophan, which is required for chlamydial growth. Other immunomodulating agents, including interleukin-1 (IL-1), can interact synergistically with IFNs, resulting in increased IDO activity in macrophages. The objectives of this study were to establish that IL-1 can enhance IFN-mediated inhibition of chlamydial growth by increasing the amount of IDO activity induced by IFNs and to identify immunomodulatory agents in culture supernatants from chlamydia-infected macrophages that interact synergistically with IFNs in restricting chlamydial growth. Monocyte-derived macrophages were treated with IL-1 combined with gamma IFN (IFN-gamma) or IFN-beta. The ability of treated cells to support the growth of Chlamydia psittaci was directly related to the amount of IDO activity induced; as IDO activity increased, so did inhibition of chlamydial growth. Furthermore, concentrations of IFNs were identified at which little IDO activity was induced and chlamydial growth was permitted yet which in the presence of IL-1 resulted in increased IDO activity and restriction of chlamydial growth. The addition of exogenous tryptophan reversed the effect of combined IFN and IL-1 treatment, indicating that IDO activity induced by combined cytokine treatment was responsible for chlamydial inhibition. Supernatants from chlamydia-infected macrophages were capable of potentiating IDO induction by IFN-gamma and of restricting the growth of C. psittaci. Antibody to IL-1 beta neutralized the potentiating effects of supernatants from chlamydia-infected cells on both IDO induction and chlamydial inhibition. Thus, IL-1 produced in response to chlamydial infection may contribute to the elimination of the infection.  相似文献   

14.
P L Atreya  S Kulkarni 《Virology》1999,261(2):227-241
Respiratory syncytial virus (RSV) belongs to Paramyxoviridae family of enveloped negative-strand RNA viruses and causes severe bronchiolitis and pneumonia in children younger than 2 years of age. As members of Paramyxoviridae family, RSV and parainfluenza type 3 (PIV3) have similar modes of infection and replication. A variety of negative-strand RNA virus infections, including that of PIV3, are inhibited by human MxA protein, a type I interferon (IFN)-inducible GTPase. We tested whether the MxA protein, induced either by type I human IFNs or by stable transfection of human MxA gene in human (U-87) or simian (Vero) cells, confers resistance to these cells against infection by RSV strain A2. RSV infection was resistant to antiviral effects induced by 0-10,000 U/ml type I IFNs (IFN-alpha or -beta) in both human lung epithelial, A549, and fibroblast, MRC-5 cells. RSV virus yield was reduced only by 10- to 20-fold, and viral protein synthesis was not significantly affected under conditions of IFN treatment where PIV3 yield was reduced by 1000- to 10,000-fold. Human or simian cell lines constitutively expressing MxA were protected against infection by PIV3 but not by RSV. Our results indicate that RSV A2 is resistant to the antiviral effects of MxA, even though RSV and PIV3 have similar replication strategies. In IFN-treated coinfected cultures, IFN-resistant RSV A2 did not prevent the IFN-mediated inhibition of PIV3 multiplication. Hence the resistance of RSV A2 to type I IFNs does not appear to be due to soluble factors released into the medium or a disruption in the cellular antiviral machinery brought about by RSV A2 infection.  相似文献   

15.
The influence of pretreatment with interferon (IFN) on subsequent IFN synthesis was investigated in macrophage cultures of DBA/2 and C57BL/6 mice. The doses of IFN alpha/beta for pretreatment ranged from 10,000 U/ml to 100 U/ml and the incubation time was between 18 and 2 h. No blocking effect was observed for chemical induction with poly I:poly C or CMA. However, for viral infection with NDV, blocking was observed. This inhibition of IFN synthesis was dependent on the dose and time of IFN pretreatment and of the titer of the inducing virus. Similarly in mouse fibroblast cultures no blocking activity was observed for induction with poly I:poly C/DEAE-dextran. Again, with NDV as inducer, pretreatment with IFN resulted in inhibition of interferon synthesis. Thus, our data show that blocking occurs only with a viral inducer and suggest that it is caused by an antiviral effect.  相似文献   

16.
To investigate the possible interactions between human and porcine interferons (IFNs) in vitro, human transformed (FL) and nontransformed (HEF) cells were treated with either HuIFN alpha and/or gamma and porcine alpha and/or gamma. In both cases the antiproliferative activity was measured to determine the effects of different combinations between human and porcine IFNs on cell proliferation. Combinations of human and porcine IFNs acted mostly antagonistically with exception of IFN combination Hu-alpha/Po-gamma which showed a synergic cooperativity in therms of antiproliferative activity on human transformed cells.  相似文献   

17.
18.
In view of the immunoregulatory and antiviral properties of the interferons (IFNs), the production of and response to these cytokines in vivo and in vitro were assessed in 42 patients with multiple sclerosis (MS), a disease with features of autoimmunity and a viral infection. Serum IFN, determined by bioassay of antiviral activity at 10 intervals over 18 months, was detectable at levels ranging from 16 to 250 IU/ml, at least once and up to five times in 37 of the 42 patients. Of 420 samples tested, 88 (21%) were positive. None of the 71 serum samples from 37 healthy subjects contained detectable IFN activity. Neutralization of antiviral activity by antibodies showed that the serum IFN type was IFN-alpha in 82 samples, IFN-gamma only in 2, and both IFN-alpha and IFN-gamma were present in 4. At the initial time point the activity of 2'-5' oligoadenylate synthetase (OAS), an IFN-induced enzyme, was elevated in peripheral blood leucocytes (PBL) from 13 patients, but not in 7 patients seropositive for IFN, indicating that in some patients there was a failure of PBL to respond to endogenous IFN. In most patients the capacity of PBL in vitro to produce IFNs-alpha/beta or -gamma after induction by virus or mitogens, respectively, was likewise reduced. These various abnormalities in IFN responses could not be correlated with clinical assessments of disease activity but may reflect subclinical attacks. The abnormalities described, in particular the intermittent interferonemia in MS, are more striking than in other diseases previously reported, indicating an unusual component to the stimulus for IFN production (viral or other) or the response to it. The effects of endogenous IFN production may have implications for the scheduling of therapy with IFN in MS.  相似文献   

19.
Bacteria-derived human leukocyte interferon (IFN) subtypes, IFN-alpha A, -alpha B, and -alpha D, and two hybrid IFNs, IFN-alpha AD and -alpha DA, were examined for both in vitro and in vivo antiviral activity. Two of these materials in highly purified form (IFN-alpha D and -alpha D) protect mice against lethal doses of encephalomyocarditis virus infection. A single dose of 1 microgram of protein of IFN-alpha D 3 h before infection conferred protection in both BDF1 and CD-1 mice against encephalomyocarditis virus infection, and multiple treatments with IFN-alpha D or IFN-alpha AD extend the mean survival time of infected mice. On a weight basis, IFN-alpha AD was approximately 100-fold more effective than IFN-alpha D. There is a direct correlation between the antiviral activity of the various human IFN species in L-929 cells and in mice for both single and multiple treatments before infection, but none of the cloned human IFN subtypes were effective when administered 24 h after infection. Mixtures of the two parental materials, IFN-alpha A and -alpha D, were not as protective as the hybrid molecule IFN-alpha AD, suggesting that IFNs with unique and altered species specificity can be produced by recombinant DNA methods.  相似文献   

20.
MuIFN-alpha/beta at a concentration of 0.5 IU/ml exerted both antiviral and priming effects in the B, but not in the M subline of L929 cells. This concentration of IFN did not affect the small size 2-5A synthetase mRNA levels in either sublines, as determined by a Northern blot technique. Pretreatment with 5 or 50 IU/ml IFN resulted in more pronounced antiviral and priming effects in the L929B than in the L929M cells. However, the enhancements of the 2-5A synthetase mRNA levels due to the pretreating IFN dose were similar in both sublines.  相似文献   

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