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1.
The relationship between growth and interferon- (IFN-) production in the recombinant cell line CHO 320 was studied by varying the foetal calf serum (FCS) concentration. The specific growth rate varied with the initial FCS concentration in a manner which could be well fitted by the Monod model. TheK
s and
max
values were found to be 0.771% (v/v) serum and 0.031 h–1 respectively. The average specific IFN- production rates during the exponential phase increased with increasing FCS concentration. A good correlation between specific production rate and specific growth rate was found in all phases of the culture except the lag phase and it was clearly demonstrated that IFN- production was growth associated. Specific glucose and glutamine utilisation rates were inversely related to specific growth rates. 相似文献
2.
Joseph A. Trapani Kylie A. Browne Michelle J. Dawson Robert G. Ramsay Roger L. Eddy Thomas B. Shows Perrin C. White Bo Dupont 《Immunogenetics》1992,36(6):369-376
A cluster of at lest six interferon- (IFN)-inducible genes designated Ifi201-204 and located on mouse chromosome 1 has recently been described. Here , we report a human IFN--inducible gene, IFI 16, which has nucleotide sequence similarity with portions of two of the mouse genes, Ifi202 and Ifi204. A full-length cDNA clone derived from IFI 16 [2.709 kilobases (kb)] contained a single open reading frame of 2.187 kb which encoded a putative polypeptide of 729 amino acids and a predicted non-glycosylated M
r
of 80020. IFI 16 mRNA was found to be constitutively expressed in lymphoid cells and in cell lines of both the T and B lineages. By contrast, the mRNA was not expresed by the cell lines HL-60, U937, and K562, which represent early stages of myeloid development, but was strongly inducible in HL-60 and U937 with IFN-. The IFI 16 protein demonstrated a putative domain structure with patchy similarity to the proteins expressed from gene Ifi202 and Ifi204. The mouse and human proteins each contain two analogous 200 amino acid domains which are imperfect copies, but IFI 16 demonstrated additional unique regions, including a Lys-rich N-terminal portion and a spacer region between the reiterated domains, analogous to spacer regions in the CD5 and CD8 molecules. Using a panel of inter-species somatic cell hybrid cell lines, IFI 16 was localized to the chromosomal region 1q121qter, a region systenic between mouse an man. DNA blotting indicated that, in contrast to the mouse, IFI 16 is present as a single copy gene in the human genome.The authors are pleased to make the cDNA clones described in this paper available to interested investigators.The nucleotide sequence data reported in this paper have been submitted to the Genbank database and have been assigned the accession number M63838.
Correspondence to: J. A. Trapani. 相似文献
3.
Satoh R Kotake M Takano T Motokawa K Gemma T Watanabe R Arai S Hohdatsu T 《Microbiology and immunology》2011,55(3):184-190
Unmethylated CpG-ODN are known to enhance Th1-type immune response. However, optimal sequences of CpG-ODN for activating Th1-type immune cells vary among species. It is necessary to identify the effective CpG-ODN sequences in each species. In the present study, in order to identify the sequences of CpG-ODN that produce fIFN-γ in cats, 14 kinds of ODN were synthesized and examined regarding their ability to induce fIFN-γ in feline PBMC and splenocytes. It was shown that some CpG-ODN significantly induced fIFN-γ production in splenocytes, but not in PBMC. We found that three kinds of CpG-ODN (no. 2, 5'-ggTGCATCGATGCAGggggG-3'; no. 5, 5'-ggTGCGTCGACGCAGggggG-3'; no. 10, 5'-ggTGCTACGTAGCAGggggG-3') specifically and significantly induced fIFN-γ production in feline splenocytes. The reverse sequences, GpC-ODN, do not cause significant fIFN-γ production. The fIFN-γ production inductivity of a mixture of CpG-ODN nos. 2, 5 and 10 was higher than those of individual CpG-ODN. When the CpG-ODN mixture was encapsulated in an MCL and administrated to cats, the number of fIFN-γ(+) cells in PBMC significantly increased. CpG-ODN nos. 2, 5 and 10 should be useful to elicit a Th1-type immune response as a vaccine adjuvant in cats. 相似文献
4.
Fenfen Zhu Qi Wang Hefang Pu Shasha Gu Lan Luo Zhimin Yin 《World journal of microbiology & biotechnology》2013,29(2):319-325
Interferon-γ (IFN-γ) is a broad-spectrum antiviral glycoprotein that produced by lymphatic T cells and natural killer cells those who had stimulated by antigen. Human IFN-γ (hIFN-γ) often used in clinical research and practice because of its bioactivity, for example, antivirus, antitumor, controlling cell apoptosis, and the strict selectivity. However, due to the difficulties of Escherichia coli expression system meet in protein folding, the hIFN-γ often existed as inclusion body. The production of soluble hIFN-γ can be developed to shorten the production cycle and decrease the cost. In this study, small ubiquitin-related modifier fusion technology was used to express and purify recombinant hIFN-γ. Expression induced by adding 50 mM arginine and 1 % (w/v) glycerol into the culture at 24 °C existed as a soluble form of 70 % in total protein. Finally, about 62 mg recombinant hIFN-γ was obtained from 1 L fermentation culture with no less than 96 % purity. Determined by cytopathic effect inhibition assay, the specific activity of the recombinant hIFN-γ achieved at 7.78 × 105 IU/mL. 相似文献
5.
Andreas von Knethen Lisa K. Sha Laura Kuchler Annika K. Heeg Dominik Fuhrmann Heinrich Heide Ilka Wittig Thorsten J. Maier Dieter Steinhilber Bernhard Brüne 《Cellular signalling》2013,25(12):2762-2768
Macrophage polarization to an anti-inflammatory phenotype upon contact with apoptotic cells is a contributing hallmark to immune suppression during the late phase of sepsis. Although the peroxisome proliferator-activated receptor γ (PPARγ) supports this macrophage phenotype switch, it remains elusive how apoptotic cells activate PPARγ. Assuming that a molecule causing PPARγ activation in macrophages originates in the cell membrane of apoptotic cells we analyzed lipid rafts from apoptotic, necrotic, and living human Jurkat T cells which showed the presence of 5-lipoxygenase (5-LO) in lipid rafts of apoptotic cells only. Incubating macrophages with lipid rafts of apoptotic, but not necrotic or living cells, induced PPAR responsive element (PPRE)-driven mRuby reporter gene expression in RAW 264.7 macrophages stably transduced with a 4xPPRE containing vector. Experiments with lipid rafts of apoptotic murine EL4 T cells revealed similar results. To verify the involvement of 5-LO in activating PPARγ in macrophages, Jurkat T cells were incubated with the 5-LO inhibitor MK-866 prior to induction of apoptosis, which failed to induce mRuby expression. Similar results were obtained with lipid rafts of apoptotic EL4 T cells preexposed to the 5-LO inhibitors zileuton and CJ-13610. Interestingly, Jurkat T cells overexpressing 5-LO failed to activate PPARγ in macrophages, while their 5-LO overexpressing apoptotic counterparts did. Our results suggest that during apoptosis 5-LO gets associated with lipid rafts and synthesizes ligands that in turn stimulate PPARγ in macrophages. 相似文献
6.
The importance of cell cycle analysis in cell culture development has been widely recognised. Whether such analysis is useful
in indicating future performance of high cell density culture is uncertain. Using flow cytometric approach to address this
question, we utilised the fraction of cells in the S phase to control specific growth rate and productivity in spin filter
perfusion cultures and found a significant increase in the accumulated interferon-γ over that obtained from the nutrient-based
controlled fed culture. While a general decrease with time exists in both percentage of S phase cells and specific growth
rate, a clear oscillatory behaviour of both parameters is found in perfusion cultures.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
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Heidari-Japelaghi Reza Valizadeh Mostafa Haddad Raheem Dorani-Uliaie Ebrahim Jalali-Javaran Mokhtar 《Transgenic research》2020,29(4):381-394
Transgenic Research - The plant-based expression systems are now accredited as bioreactors for the high production of various biopharmaceuticals. However, low levels of agglomeration and the... 相似文献
11.
Sungjin Ahn Moonyoung Lee Seungchan An Sooyeol Hyun Jiho Hwang Jongkook Lee Minsoo Noh 《Bioorganic & medicinal chemistry》2018,26(5):1069-1075
Adiponectin is a major adipocytokine secreted from mammalian adipocytes. Relatively low expression of adiponectin is associated with various human metabolic diseases and some cancers. Adiponectin-secreting compounds have therapeutic potential for these diseases. Adipogenesis of human bone marrow-mesenchymal stem cells (hBM-MSCs) has been used as a phenotypic assay to find adiponectin secreting compounds. In a phytochemical library screen, 2-formyl-komarovicine, 1-(quinolin-8-yl)-1,3,4,9-tetrahydro-2H-pyrido[3,4-b]indole-2-carbaldehyde, isolated from Nitraria komarovii was identified as a potential adiponectin-secreting compound. To validate the results of the impure phytochemical, we synthesized 2-formyl-komarovicine. The synthetic 2-formyl-komarovicine significantly promoted adiponectin production during adipogenesis in hBM-MSCs. In a target identification experiment, 2-formyl-komarovicine bound to peroxisome proliferator-activated receptor γ (PPARγ) in a concentration-dependent manner. Notably, 2-formyl-komarovicine competitively inhibited the adiponectin-promoting activity of a full PPARγ agonist, troglitazone, in hBM-MSCs, which is a pharmacological feature of a partial agonist. The ligand-docking model showed that 2-formyl-komarovicine interacted with the hydrophobic pocket of the PPARγ ligand-binding domain, but lacked an interaction to stabilize helix H12, which is one of the major binding themes of PPARγ partial agonists. We concluded that 2-formyl-komarovicine provides a novel pharmacophore for PPARγ partial agonists to increase adiponectin production. 相似文献
12.
The effects of lipids on the glycosylation of recombinant human interferon- expressed in a Chinese Hamster Ovary cell line were investigated in batch culture. Lipids form an essential part of the N-glycosylation pathway, and have been shown to improve cell viability. In control (serum-free) medium the proportion of fully-glycosylated interferon- deteriorated reproducibly with time in batch culture, but the lipoprotein supplement ExCyte was shown to minimise this trend. Partially substituting the bovine serum albumin content of the medium with a fatty-acid free preparation also improved interferon- glycosylation, possibly indicating that oxidised lipids carried on Cohn fraction V albumin may damage the glycosylation process.Abbreviations BSA
bovine serum albumin
- CHO
chinese hamster ovary
- DHFR
dihydrofolate reductase
- FCS
foetal calf serum
- IFN-
human interferon-gamma
-
q
IFN
specific interferon production rate
-
specific growth rate
- 2N
doubly-gycosylated
- 1N
singly-glycosylated
- ON
non-glycosylated 相似文献
13.
Functional genomics of PPAR-γ in human immunomodulatory cells 总被引:1,自引:0,他引:1
Keeping in view the fact that peroxisome-proliferators activated receptors-PPARs (α,γ) play a crucial role in atherogenic
inflammation, the present study was addressed to explore as to how selective and specific PPAR-γ gene silencing within human
mononuclear cells affects genes involved in lipid metabolism and innate immune process. Such a study revealed that with respect
to control cells, the PPAR-γ knock-out cells exhibited significant reduction in the expression of genes coding for PPAR- α,
CD-36, LDL-R as well as significant increase in the expression of genes coding for IL-4, IL-8, IFN-γ, CX3CR1, hTERT. However,
the expression of genes coding for LXR-α and Receptor-C
k
could not be detected in PPAR-γ knock-out cells. Based on these results, we propose that PPAR-γ gene has the inherent capacity
to influence the lipid mediated inflammation process in atherosclerotic lesions. 相似文献
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Effect of interferon-γ on the susceptibility to Fas (CD95/APO-1)-mediated cell death in human hepatoma cells 总被引:9,自引:0,他引:9
Shin EC Shin WC Choi Y Kim H Park JH Kim SJ 《Cancer immunology, immunotherapy : CII》2001,50(1):23-30
Many tumors, including hepatocellular carcinomas (HCCs), resist Fas-mediated cell death, which is one of the effector mechanisms
in the host's anti-tumor response; however, this resistance can be abolished by interferon-γ (IFN-γ). IFN-γ may sensitize
Fas-mediated cell death in several ways, but the exact mechanism in HCCs is uncertain. In this study, we thoroughly investigated
the effect of IFN-γ on the susceptibility of one human normal liver cell line and 12 HCC cell lines to Fas-mediated cell death.
We also investigated the effect of IFN-γ on the expression of various apoptosis-related genes such as the Fas/TNF-related
genes, the bcl-2 family, and the caspase family of genes. Although most cell lines showed considerable constitutive expression of Fas, all
tested cell lines resisted Fas-mediated cell death without IFN-γ. When cells were pretreated with IFN-γ, only three cell lines
were made significantly susceptible to Fas-mediated cell death (SNU-354, SNU-387 and SNU-423); the other 10 cell lines were
not affected. IFN-γ increased the mRNA expression of Fas, TRAIL and caspase-1, and surface Fas was also increased. The strongly
sensitized cell lines (SNU-354, SNU-387 and SNU-423) showed a particularly potent increment in surface Fas after IFN-γ treatment
(increase in surface Fas >1.7-fold). This result enabled us to conclude that a potent increment of surface Fas expression
is a major sensitizing mechanism of IFN-γ. We conclude that IFN-γ cannot play a sensitizing role in most HCC cell lines and
that IFN-γ makes HCC cells susceptible to Fas-mediated cell death through a marked up-regulation of surface Fas in some HCC
cells.
Received: 3 August 2000 / Accepted: 24 November 2000 相似文献
17.
Hsp70B’ was expressed on the surface of HT-29 and CRL-1809 but not SW-480 human colon cell lines in response to proteasome
inhibition as detected using flow cytometry. Surface expression was not detected under non-stress conditions nor was heat
shock an inducer of surface expression in the three cell lines tested. Phylogenetic analysis indicated that the Hsp70B’ protein
sequence was most closely related to another major inducible human Hsp70, Hsp72. Hsp70B’ appeared to be recently diverged,
as homologs for Hsp70B’ have not been found in rodents. Hsp72 and Hsp70B’ shared 100% amino acid sequence identity in their
predicted peptide-binding regions suggesting that they bind the same peptide substrates, perhaps in extracellular antigen
presentation. Amino acid sequence differences were concentrated in the lid regions and the C-terminal domains raising the
possibility that Hsp72 and Hsp70B’ bind different co-chaperones or cell surface receptors. 相似文献
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Isidoro González-Álvaro Carmen Domínguez-Jiménez Ana M Ortiz Vanessa Núñez-González Pedro Roda-Navarro Elena Fernández-Ruiz David Sancho Francisco Sánchez-Madrid 《Arthritis research & therapy》2006,8(4):R88-11
We have characterized the lymphocyte subset and the receptor molecules involved in inducing the secretion of TNF by monocytic
cells in vitro. The TNF secreted by monocytic cells was measured when they were co-cultured with either resting or IL-15-stimulated lymphocytes,
T cells, B cells or natural killer (NK) cells isolated from the peripheral blood of healthy subjects and from the synovial
fluid from patients with inflammatory arthropathies. Co-culture with IL-15-activated peripheral blood or synovial fluid lymphocytes
induced TNF production by monocytic cells within 24 hours, an effect that was mainly mediated by NK cells. In turn, monocytic
cells induced CD69 expression and IFN-γ production in NK cells, an effect that was mediated mainly by β2 integrins and membrane-bound IL-15. Furthermore, IFN-γ increased the production of membrane-bound IL-15 in monocytic cells.
Blockade of β2 integrins and membrane-bound IL-15 inhibited TNF production, whereas TNF synthesis increased in the presence of anti-CD48
and anti-CD244 (2B4) monoclonal antibodies. All these findings suggest that the cross-talk between NK cells and monocytes
results in the sustained stimulation of TNF production. This phenomenon might be important in the pathogenesis of conditions
such as rheumatoid arthritis in which the synthesis of TNF is enhanced. 相似文献