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1.
The presence of serum in suspensions of Sendai-induced human leukocytes is necessary for the synthesis of significant amounts of interferon. Very little interferon is obtained from serum-free suspensions. Cow's milk or milk casein can substitute for serum in the production of high yields of human leukocyte interferon.  相似文献   

2.
Gamma interferon (IFN-gamma)-mediated indoleamine-2,3-dioxygenase (IDO) activity in human astrocytoma cells and in native astrocytes was found to be responsible for the inhibition of herpes simplex virus replication. The effect is abolished in the presence of excess amounts of L-tryptophan. Both IFN-alpha and IFN-beta restricted herpes simplex virus replication in both cell types, but (in contrast to the results seen with IFN-gamma) the addition of an excess amount of L-tryptophan did not inhibit the induced antiviral effect.  相似文献   

3.
The possible pathways for realization of antiviral activity of interferon inducer poly (I).poly(C) have been studied. The stimulating effect of interferon inducer on the net protein synthesis in human M19 fibroblasts has been demonstrated. Compositions of the specific proteins induced by poly(I).poly(C) or interferon in human M19 fibroblasts and in monkey cells 4647 have been analyzed by electrophoresis technique. The data obtained suggest the existence of common gene products for interferon and ds-inducer. The ds-inducer requires the synthesis of lesser amounts of proteins for realization of its biological activity as compared with interferon.  相似文献   

4.
The results of using human interferon of types I (alpha- and beta-interferons) and II (gamma-interferon) in combination with chemotherapeutic drugs, such as remantadin and ribavirin for the study in human cell cultures are presented. Moderate doses of the drugs (25 microgram/ml) did not eliminate the interferon production. In higher doses (50 microgram/ml) they lowered the interferon production levels 2--3 times. In the presence of ribavirin the level of the interferon production lowering was higher. On the whole the effect of the drugs on production of interferons of types I and II was of a similar character despite the different means of interferon induction. The combined use of interferons of types I and II with the chemotherapeutic drugs in human embryo cultures infected with Semiliki forest virus (SFV) revealed an additive character of the antiviral effect in all combinations tested. The level of the antiviral activity of alpha-, beta- and gamma-interferons against the SFV was practically the same.  相似文献   

5.
Production and characterization of interferon from endothelial cells   总被引:3,自引:0,他引:3  
The capacity of cultured bovine aortic, capillary, and corneal endothelial cells as well as of human umbilical cord endothelial cells to produce interferon (IFN) was investigated. The endothelial cells of the two species produced significant amounts of IFN in response to various viruses and poly (I) poly (C). The IFN produced by human umbilical cord endothelial cells was a mixture of alpha- and beta-IFN, as determined by neutralization with antibodies directed against these two types of IFNs as well as by measuring the antiviral activity on heterologous cells. Bovine endothelial cells also produced a mixture of at least two IFN subspecies, one of them labile at pH 2 and active on human cells and the other stable at pH 2 and inactive on human cells.  相似文献   

6.
Current advances in the use of somatic cell hybrid systems have enhanced the value of these systems for studying eukaryotic cell functions. We have reviewed the use of somatic cells to investigate the human interferon system. It has been shown that interspecific heterokaryons and hybrid cells can produce interferon(s) of both parental types and may be protected from viral challenge by interferon(s) from either parent. Using mouse-human hybrid cells we have assigned a human gene(s) responsible for regulating interferon to chromosome 21 and genes involved in the production of human interferon to chromosomes 2 and 5. Our data also suggest possible assignment of a locus involved in control of interferon production to chromosome 16. Suggested further uses of the somatic cell system for interferon studies include study of the subunit structure of interferons and the development of hybrid lines that produce human interferon at high levels (interferon/somatic cell hybrids/human gene assignment.  相似文献   

7.
Bacterial synthesis of a novel human leukocyte interferon.   总被引:11,自引:5,他引:6       下载免费PDF全文
A novel human leukocyte interferon cDNA clone (LeIF B) was identified in a cDNA library prepared using polyadenylated mRNA of a myeloblastoid cell line. The nucleotide sequence of LeIF B differs significantly from other published leukocyte interferon cDNA sequences. An expression plasmid was constructed which directs the synthesis in E. coli of 8 x 10(7) interferon units per liter of culture. LeIF B exhibits markedly different specificities from another bacterially synthesized human leukocyte interferon, LeIF A.  相似文献   

8.
The carbohydrate content of all of the species of human leukocyte interferon (IFN-alpha) which have been derived from patients with chronic myelogeneous leukemia (CML) and purified to homogeneity has now been determined. Amino sugar content was measured by high-performance liquid chromatography and fluorescamine detection of acid hydrolysates of each sample. Two species showed significant amounts of glucosamine. Most of the purified species of leukocyte interferon from a myeloblast cell line were also tested, and two species were found to contain sugar residues. These forms also differed from the CML interferons in that they revealed the presence of greater amounts of galactosamine. The apparent lack of carbohydrate in some of the higher-molecular-weight species of interferon implicated factors other than glycosylation in the molecular weight differences. The results indicate that some species of IFN-alpha are glycosylated to various degrees.  相似文献   

9.
Microassay for Human and Chick Cell Interferon   总被引:15,自引:2,他引:13       下载免费PDF全文
The microassay for human and chick interferon described in this paper required much smaller amounts of samples and reagents and considerably less time and effort than the plaque reduction assay while yielding comparable results.  相似文献   

10.
干扰素调节免疫具有广谱抗病毒和抗肿瘤作用。重组人干扰素α1b是国内自主产权的基因工程I类新药,通过多年研究,研制出儿童和成人应用的重组人干扰素α1b注射液、喷雾剂等多种剂型,完成治疗冠状病毒等肺炎的药理学、毒理学、药代动力学和相关临床研究,形成了多个临床应用的专家共识和指南性文件。新型冠状病毒肺炎疫情发生后,这些研究为国家卫健委将干扰素α雾化吸入作为首推抗病毒药物试用方案纳入《新型冠状病毒感染的肺炎国家诊疗方案》提供了基础。经过大量新冠肺炎患者的临床使用,取得了良好的治疗效果。重组人干扰素α1b喷雾剂作为可预防病毒感染的药物,用于医护人员等高暴露人群的生物防护,为疫情防控发挥了重要作用,将对国民健康和国家生物安全保障做出应有的贡献。  相似文献   

11.
New simple dye-uptake assay for interferon   总被引:1,自引:0,他引:1  
Using the spectrophotometer that the authors developed, the amounts of human leukocyte and mouse L cell interferons on FL cells and L929 cells were measured and values were compared with those measured by the cytopathogenic effect (CPE) reduction method (CPE method). The spectrophotometric method, which was simpler than the original dye-uptake method, was found to be more sensitive than the latter. When Sindbis virus was used instead of vesicular stomatitis virus (VSV), there were no significant differences in the sensitivities of the two methods or the interferon titers estimated. When FL cells or L929 cells were treated with interferon at the time of their dispersion, their interferon titers were almost the same as those of cells treated with interferon 2 days after dispersion. It is concluded that this new dye-uptake method is useful for assay of human and mouse interferons.  相似文献   

12.
Studies were done to determine whether differences in interferon production are responsible for the resistance of pastel mink to Aleutian disease. The abilities of normal pastel and sapphire mink to produce interferon when inoculated with either Newcastle disease virus or a synthetic polyribonucleotide, poly (I):poly (C), were identical, even to the production of a novel, acid-labile interferon. The resistance of pastel mink to Aleutian disease did not correlate with interferon production, because neither sapphire nor pastel mink produced detectable amounts of interferon when infected with either the Pullman strain of Aleutian disease virus (ADV) or the highly virulent Utah I strain. Sapphire mink infected with the Pullman strain responded normally to poly (I):poly (C) early in the course of the disease, but interferon production was impaired late, when the mink were hypergammaglobulinemic and had renal, vascular, and hepatic lesions. These data suggest that ADV Pullman neither stimulates nor interferes with interferon production in infected mink and may represent a mechanism whereby ADV can more readily establish infection.  相似文献   

13.
Interferon (IFN) production by human peripheral lymphocytes stimulated with M. pneumoniae was investigated. The hydrogen peroxide released from M. pneumoniae was responsible for the induction of IFN from lymphocytes, since horseradish peroxidase inhibited the IFN production and abrogated the activity of IFN production in the supernatant of M. pneumoniae. The antiserum neutralizing IFN alpha and IFN beta failed to neutralize partially interferon produced by lymphocytes. Treatment either with pH 2.0 or antiserum neutralizing human IFN gamma resulted in a partial reduction of interferon. These results indicate that interferon produced by human lymphocytes stimulated with M. pneumoniae includes both types of IFN gamma and IFN beta.  相似文献   

14.
Kang D  Jiang H  Wu Q  Pestka S  Fisher PB 《Gene》2001,267(2):233-242
Defects in growth control and differentiation occur frequently in human cancers. In the case of human melanoma cells, treatment with a combination of fibroblast interferon (IFN-beta) and the protein kinase C activator mezerein (MEZ) results in an irreversible loss of proliferative potential and tumorigenic properties with a concomitant induction of terminal differentiation. These changes in cellular properties are associated with an induction and suppression in specific subsets of genes that occur in a temporal manner. To identify the complete repertoire of gene changes occurring during melanoma reversion to a more differentiated state a number of molecular approaches are being used. These include, subtraction hybridization using temporally spaced cDNA libraries, random cDNA isolation and evaluation by reverse Northern blotting and high throughput microarray analysis of subtracted cDNA clones. In the present study we have used a novel approach, rapid subtraction hybridization (RaSH), to identify and clone an additional gene of potential relevance to cancer growth control and terminal cell differentiation. RaSH has identified a human ubiquitin-processing protease gene, HuUBP43, that is differentially expressed in melanoma cells as a function of treatment with IFN-beta or IFN-beta + MEZ. HuUBP43 is a type I interferon inducible gene that is upregulated in a diverse panel of normal and tumor cells when treated with IFN-beta via the JAK/STAT kinase pathway. This gene may contribute to the phenotypic changes induced by IFN-beta during growth arrest and differentiation in human melanoma cells and other cell types as well as the antiviral and growth inhibitory effects of interferon.  相似文献   

15.
Interferons obtained on induction of human lymphocytes with Newcastle viruses and staphylococcal enterotoxin A and diploid fibroblast cells of human embryos with poly (I).poly (C), as well as translation products of interferon mRNA obtained from these cells were analysed serologically. It was shown that the main type of interferon produced by the cells depended on the cell culture and inductor nature. It was defined at the level of the respective gene depression. Effective translation of mRNA of the interferons of the 3 types makes possible production of cDNA and creation of bacterial plasmids coding the genetic information for the synthesis of human interferon.  相似文献   

16.
Interferons are naturally occurring proteins that are currently under evaluation as potential antiviral and antitumor agents. Currently all human interferons can in principle be produced in adequate amounts by recombinant DNA technology. Human interferons produce side effects, but because they are species-specific the toxicity cannot be tested in lower mammals. The chimpanzee is the only species in which the side effects of human interferon can be reproduced, and only in this species the toxicity of human interferons can be screened.  相似文献   

17.
Johnson, Terry C. (University of Minnesota, Minneapolis), and Leroy C. McLaren. Plaque development and induction of interferon synthesis by RMC poliovirus. J. Bacteriol. 90:565-570. 1965.-Plaque development by RMC poliovirus on human amnion cell monolayers was investigated with regard to autointerference and to the effect of acid-agar overlay on plaquing efficiency. The virus was inhibited by acid-agar overlay, thereby exhibiting the d(-) marker typical of attenuated poliovirus strains. In addition, a lack of RMC poliovirus plaque development on HeLa cell monolayers was shown to be the result of an agar inhibitor which could be removed by NaCl extraction. By use of a simplified plaque reduction assay, it was shown that interferon production was responsible for the autointerference phenomenon. Interferon synthesis did not correlate with the ages in vitro of human amnion cell cultures. Fibroblasts originating from the chorionic membrane produced negligible amounts of the inhibitor. Interferon synthesis by human amnion cells infected with RMC poliovirus was inhibited by actinomycin D. The addition of guanidine hydrochloride to infected cultures immediately after RMC poliovirus adsorption markedly inhibited interferon synthesis, although after 2 hr (postadsorption) guanidine had no effect on interferon production.  相似文献   

18.
The preparation is described of three types of liposomes containing biologically active human fibroblast interferon. Depending on the preparative method, up to 50% of the initial interferon activity could be recovered associated with the liposomes, 15–30% being entrapped into the aqueous space of the vesicles. Encapsulation into negatively charged liposomes is dependent on the acidic phospholipid content; liposomes bearing a net positive charge could capture more interferon than those with a negative charge but were toxic to the target cells. Expression of biological activity of liposomes encapsulated interferon was demonstrated by their antiviral activity and their ability to prime interferon induction.  相似文献   

19.
Antiviral and cell-growth-inhibitory activities of human interferon were shown to be related to the activity of a gene or genes present on chromosome 21. The 18s rRNA is vital to cell growth; it is capable of a viral-mRNA-recognition function and it is coded for by genes a portion of which are present on chromosome-21. A previously reported ability of human interferon to affect rRNA metabolism is characterized by a decrease in the sucrose-gradient-peak ratio of radiolabelled 28S to 18S rRNA in extracts from the cytoplasm of interferon-treated human fibroblasts. In the present report, interferon dose-response curves are presented demonstrating a direct relationship between a decrease in this ratio and interferon concentrations in the media. By using this virus-independent cytoplasmic rRNA assay, eight human fibroblast lines, differing in chromosome 21 ploidy, were tested for sensitivity to human interferon. Two monosomy-21, two euploid-21 and four trisomy-21 cell lines were tested. The monosomy-21 cell populations were significantly less sensitive to interferon than the other six cell types tested. Of the cell lines tested, the most sensitive, by a wide margin, was a trisomy-21 line. Trisomy-21 cell monolayer sensitivity, however, varied widely within the range from normal to supersensitive. These observations suggest that interferon's ability to affect rRNA metabolism is related to the activity of a gene or genes present on chromosome 21.  相似文献   

20.
Two lines of Friend virus (FV)-transformed mouse spleen cells have been analyzed in respect to their interferon production capacity: neither F4 cells, which liberate infectious FV when kept under tissue culture conditions, nor the thymidine kinase-deficient B8 cells, which do not produce significant amounts of FV, release detectable amounts of autogenous interferon into cell supernatants. However, interferon is produced in these cells in amounts comparable to that in L-929 cells when interferon induction is initiated with UV-inactivated Newcastle disease virus. Conversely poly(I)-poly(C), a potent interferon inducer in L-929 cells, proved ineffective in this capacity in F4 or B8 cells. When erythropoietic differentiation is induced in these cells by dimethyl sulfoxide, no autogenous interferon production occurs, but with NDV-induction a four- to fivefode increase of interferon production is observed. A similar elevation of interferon production is achieved during 5-bromodeoxyuridine stimulation of differentiation in the thymidine kinase-deficient B8 cells. The refractiveness against poly(I)-poly(C) displayed in unstimulated cells is not overcome at any stage of differentiation, indicating major differences of Newcastle disease virus and poly(I)-poly(C) induction mechanisms.  相似文献   

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