共查询到20条相似文献,搜索用时 15 毫秒
1.
N M Gough D J Hilton D P Gearing T A Willson J A King N A Nicola D Metcalf 《Blood cells》1988,14(2-3):431-442
A cDNA library was constructed using mRNA from Krebs ascites tumor cells that was shown by Northern blot hybridization to contain mRNA for murine leukemia inhibitory factor (LIF). This library was screened with an oligonucleotide corresponding to the 3' end of a partial LIF cDNA clone, and an overlapping cDNA clone isolated. Nucleotide sequence analysis of this latter clone allowed the complete sequence of LIF to be derived. A cDNA molecule encoding the entire mature LIF protein was installed in a yeast expression vector, and LIF produced up to about 100 ng/ml in the growth medium. The LIF produced by yeast cells has the same biologic properties as native LIF and competes with native 125I-LIF for binding to specific cellular receptors. Two forms of native LIF, distinguishable by their chromatographic behavior on DEAE-Sepharose, were converted by neuraminidase treatment to a form with similar chromatographic behavior, suggesting that the major difference between these two species is the content of sialic acid on the carbohydrate portion. Moreover, yeast-derived recombinant LIF appears to display a different pattern of glycosylation to both forms of native LIF. From in vitro experiments, we conclude that the nature of the glycosylation is not crucial to biologic activity. 相似文献
2.
The lymphoblastoid cell lines BLY 9.84, Gl, and Pl constitutively release a colony-stimulating activity (CSA) which specifically stimulates murine macrophage progenitor cells in vitro. The biochemical characterization of BLY 9.84-derived CSA exhibits a molecular size of apparently 150-200 kDa even under dissociating conditions with 6 M guanidine hydrochloride or inhibition of glycosylation. Production of this CSA is inhibited by cycloheximide and its activity is destroyed by reduction with dithiothreitol. Replating experiments give evidence for a functional similarity with CSF-1 (macrophage colony-stimulating factor). 相似文献
3.
Colony-stimulating factors (CSFs) produced by two simian virus 40(SV40) transformed macrophage cell lines (BAM1 and BAM3), and three hybrids (HM3-11, HM3-12, and HM3-14) derived from fusion between BAM3 and a Chinese hamster cell line (hs222-16) were examined. HM3-11 and HM3-14 produce two molecular species of CSF, which are not found in the conditioned media from cultures of BAM1 and BAM3 or lipopolysaccharide (LPS), phorbolmyristate-acetate (PMA), and zymosan-stimulated BAM3. HM3-12, which is classified into another group in terms of CSF secretion, does not produce these two CSFs. On the basis of various criteria, one of these CSF species (peak 1-CSF) was characterized as a macrophage-colony-stimulating factor (M-CSF). The other CSF (peak 2-CSF) induced a group of bone marrow cells in granulocytes and macrophages as well as growth of a mast cell line, IC2. This CSF has an apparent molecular weight of 18,000, estimated by SDS-polyacrylamide gel electrophoresis. Unlike interleukin 3 (IL3) from WEHI-3 cells, the growth factor activity of peak 2-CSF binds to DEAE-Sephacel. Thus, peak 2-CSF is similar to a granulocyte-macrophage colony-stimulating factor (GM-CSF) rather than to IL3. The anti L cell CSF serum does not inhibit the CSF activity in Chinese hamster fibroblast conditioned medium, and the IC2 cells do not respond to Chinese hamster lung conditioned medium (CHLCM), suggesting that peak 1- and peak 2-CSF are of mouse origin. 相似文献
4.
Expression of IgG Fc receptors in myeloid leukemic cell lines. Effect of colony-stimulating factors and cytokines 总被引:3,自引:0,他引:3
J L Liesveld C N Abboud R J Looney D H Ryan J K Brennan 《Journal of immunology (Baltimore, Md. : 1950)》1988,140(5):1527-1533
Three classes of FcR have been defined on human myeloid cells by their reactivity with mAb; FcRI (mAb 32); FcRII (mAb IV3); and FcRIII (mAb 3G8). We have quantitated the expression of each FcR on human myeloid leukemia cells and cell lines (KG-1, HL-60, U937, and K562). Detailed analysis of FcR surface expression is provided for the U937 cell line after exposure to CSF and cytokines. Increased expression of FcRI and FcRII occurred at 72 h in cells exposed to GCT or Mo cell line-conditioned medium as well as to medium from PHA-treated mononuclear cells. The augmentation of FcRII required protein synthesis and was diminished by a neutralizing antibody to granulocyte-macrophage CSF. We also show that fractions containing natural granulocyte CSF or granulocyte-macrophage CSF as well as r-granulocyte and r-granulocyte-macrophage CSF are capable of inducing FcRII on these cells, whereas other cytokines such as IL-1 and IL-2, TNF-alpha, INF-gamma and macrophages CSF failed to do so. 相似文献
5.
Differentiation of leukemic cell lines: a review focusing on murine erythroleukemia and human HL-60 cells 总被引:3,自引:0,他引:3
Several acute leukemia cell lines respond to chemical or pharmacologic inducing agents by undergoing variable degrees of differentiation. This review focuses on the manner in which murine erythroleukemia (MEL) and HL-60 cells can be induced to differentiate into virtually fully mature erythroid cells and mature granulocytes or macrophages respectively. In this process the cells undergo irreversible "commitment" to terminal differentiation which is followed by loss of proliferative capacity and alterations in the expression of genes whose products are related to specific aspects of cell maturation in the corresponding cell pathways. 相似文献
6.
Regulation of cell surface receptors for different hematopoietic growth factors on myeloid leukemic cells. 总被引:2,自引:0,他引:2 下载免费PDF全文
There are clones of myeloid leukemic cells which are different from normal myeloid cells in that they have become independent of hematopoietic growth factor for cell viability and growth. The ability of these clones to bind three types of hematopoietic growth factors (MGI-1GM = GM-CSF, IL-3 = multi-CSF and MGI-1M = M-CSF = CSF-1) was measured using the method of quantitative absorption at 1 degree C and low pH elution of cell-bound biological activity. Results of binding to normal myeloid and lymphoid cells were similar to those obtained by radioreceptor assays. The results indicate that the number of receptors on different clones of these leukemic cells varied from 0 to 1,300 per cell. The receptors have a high binding affinity. Receptors for different growth factors can be independently expressed in different clones. There was no relationship between expression of receptors for these growth factors and the phenotype of the leukemic cells regarding their ability to be induced to differentiate. The number of receptors on the leukemic cells was lower than on normal mature macrophages. Myeloid leukemic cells induced to differentiate by normal myeloid cell differentiation factor MGI-2 (= DF), or by low doses of actinomycin D or cytosine arabinoside, showed an up-regulation of the number of MGI-1GM and IL-3 receptors. Induction of differentiation of leukemic cells by MGI-2 also induced production and secretion of the growth factor MGI-1GM, and this induced MGI-1GM saturated the up-regulated MGI-1GM receptors. It is suggested that up-regulation of these receptors during differentiation is required for the functioning of differentiated cells. 相似文献
7.
A factor produced by feeder cells which inhibits embryonal carcinoma cell differentiation. Characterization and partial purification 总被引:2,自引:0,他引:2
Medium conditioned by STO mouse fibroblast cells inhibited both the spontaneous differentiation of NG2 embryonal carcinoma cells and the differentiation of F9 embryonal carcinoma cells induced by retinoic acid. This effect was due to a differentiation retarding factor (DRF). Reduction in DRF activity in conditioned medium by boiling and by pronase treatment suggested the involvement of a polypeptide, which had an apparent molecular weight of 57000 on gel filtration. A 28-fold purification of DRF was achieved. DRF delayed but did not prevent the extensive differentiation observed after prolonged culture of NG2 colonies. Conditioned medium could be successfully used to replace feeder cells in NG2 stock cultures. Media conditioned by a variety of other cell types also contained differentiation retarding activity. 相似文献
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9.
Kharfan-Dabaja M Ayala E Lindner I Cejas PJ Bahlis NJ Kolonias D Carlson LM Lee KP 《Cancer immunology, immunotherapy : CII》2005,54(1):25-36
Purpose: Ex vivo differentiation of myeloid leukemic blasts into dendritic cells (DCs) holds significant promise for use as cellular vaccines, as they may present a constellation of endogenously expressed known and unknown leukemia antigens to the immune system. Although variety of stimuli can drive leukemiaDC differentiation in vitro, these blast-derived DCs typically have aberrant characteristics compared with DCs generated from normal progenitors by the same stimuli. It is not clear whether this is due to underlying leukemogenic mechanisms (e.g., specific oncogenes), genetic defects, stage of maturation arrest, defects in cytokine receptor expression or signal transduction pathways, or whether different stimuli themselves induce qualitatively dissimilar DC differentiation. Methods: To assess what factors may contribute to aberrant leukemic blastDC differentiation, we have examined how the same leukemic blasts (AML and CML) respond to different DC differentiation signals—including extracellular (the cytokine combination GM-CSF+TNF-+IL-4) and intracellular (the protein kinase C agonist PMA, the calcium ionophore A23187, and the combination of PMA plus A23187) stimuli. Results: We have found that the same leukemic blasts will develop qualitatively different sets of DC characteristics in response to differing stimuli, although no stimuli consistently induced all of the characteristic DC features. There were no clear differences in the responses relative to specific oncogene expression or stage of maturation arrest (AML vs CML). Signal transduction agonists that bypassed membrane receptors/proximal signaling (in particular, the combination of PMA and A23187) consistently induced the greatest capability to activate T cells. Interestingly, this ability did not clearly correlate with expression of MHC/costimulatory ligands. Conclusions: Our findings suggest that signal transduction may play an important role in the aberrant DC differentiation of leukemic blasts, and demonstrate that direct activation of PKC together with intracellular calcium signaling may be an effective method for generating immunostimulatory leukemia-derived DCs.This work was supported by NIH CA85208, CA95829, and ASCO Young Investigator Award (M.A.K-D) 相似文献
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11.
Effects of growth rate and methotrexate on folate polyglutamates and folylpolyglutamate hydrolase activity in Krebs ascites cells 总被引:1,自引:0,他引:1
The activity of folylpolyglutamate hydrolase was measured throughout intraperitoneal growth of Krebs ascites cells in mice and after exposure to methotrexate. Hydrolase activity was lowest during the log phase of growth. Methotrexate administered intraperitoneally during log growth caused a dose- and time-dependent increase in hydrolase activity. Modest changes were observed in endogenous folate polyglutamate chain length distributions throughout growth and upon exposure to methotrexate, but these changes could not be correlated with hydrolase activity. 相似文献
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13.
Adipose cells have been recognized as an integral component of the bone marrow hematopoietic microenvironment in vivo and as an essential cell type required for in vitro maintenance of stem cells. Four stromal cell lines obtained from the adherent cell population of murine bone marrow cultures have been enriched and purified by multiple trypsinizations. We noted that these cell lines exhibited an accumulation of vacuoles of lipid, the extent of which varied be-tween cell lines in response to a change from medium containing 10% fetal calf serum to medium containing 20% horse serum. The lipid was lost when the cell lines were transferred back into the medium supplemented with fetal calf serum. In light of the reported lipogenic and antilipolytic effects of insulin on fibroblasts and adipocytes, we investigated the ability of insulin to induce adipocyte transformation of these bone marrow stromal cell populations. Three cell lines were exposed to bovine insulin at concentrations ranging from 10?9 to 10?6 M. All three cell lines responded to the insulin by accumulating lipid, but the extent of accumulation and the insulin concentration at which maximum lipid content was attained were population specific. One cell line (MC1) responded fully at physiological levels of insulin (10?9 M), whereas the other two showed lipid accumulation only at pharmacological concentrations. The initial growth of MC1 was inhibited in the presence of 10?9 M insulin which is compatible with the observed differentiation to adipocytes. The growth of MC3 was unaltered in the presence of physiological concentrations of insulin, whereas that of MC4 was accelerated. Grafts of organ cultures of the cell lines under the kidney capsule of syngeneic mice developed specific characteristics rep-resentative of the different cell lines. In particular, the majority of the grafts of MC1 consisted primarily of fat cells which were not observed in the grafts of MC3 and MC4. These data strongly suggest that these cell lines comprise cells with different potentialities and that the MC1 line represents a preadipocyte stromal cell of bone marrow. 相似文献
14.
Culture supernatants from several human leukemic T cell lines were found to contain a macrophage activating factor which enhanced hydrogen peroxide release from human peripheral blood monocyte-derived macrophages. The macrophage activating factor from a T cell line, CCRF-CEM, was characterized biochemically and compared with interferon-gamma, which is also an immunological product of T cells and has a potent macrophage activating activity. In contrast to interferon-gamma, the macrophage activating factor in the culture supernatants bound to an anion exchanger and did not adsorb onto concanavalin A gel. Culture supernatants and active fractions from chromatographies were essentially devoid of anti-viral activity. Anti-human interferon-gamma monoclonal antibody also failed to neutralize the macrophage activating factor from CCRF-CEM. MAF was eluted in the fractions with molecular weight of 40,000 to 60,000 on gel filtration in the presence of a detergent and a salt. MAF was partially purified to about 1,300-fold by the methods described above: chromatography with anion exchangers and gel filtration. It was concluded that MAF from CCRF-CEM was biochemically and immunologically different from interferon-gamma. 相似文献
15.
Interleukin-6 production by murine B cells and B cell lines 总被引:1,自引:0,他引:1
16.
1. Cell-free systems prepared from Krebs II ascites cells incubated at 45 degrees C have a much lower endogenous activity than those from cells incubated at 37 degrees C. The endogenous activity is mainly due to completion of polypeptide chains initiated in the intact cell. The low activity of the 45 degrees C system is due to a lesion in initiation in cells incubated at 45 degrees C. 2. Cell-free systems from cells incubated at 45 degrees C can translate efficiently poly (U) at 8 mM Mg2+. However, they initiate poorly on globin mRNA, indicating that these systems reflect the situation in the intact cell. 3. The lesion in globin mRNA translation in 45 degrees C systems can be overcome by addition of reticulocyte initiation factors. At saturation concentrations of factors, the response of a 45 degrees C system is restored to almost normal. 4. 45 degrees C systems from 40-S initiation complexes with methionyl tRNAfmet almost as efficiently as normal, but their ability for form 80-S complexes with globin mRNA is impaired, unless they are supplied with exogenous initiation factors. 相似文献
17.
D J McKean A Nilson B N Beck S B Mizel B S Handwerger 《Journal of immunology (Baltimore, Md. : 1950)》1985,134(4):2484-2491
The activation requirements of alloreactive and antigen reactive murine T cells were examined by stimulating class II restricted T cell clones with monoclonal B lymphoma cells. One B lymphoma cell line (T27A) was found to stimulate IL 2 release from some alloreactive T cell clones without stimulating any significant T cell proliferation response. The same B lymphoma cells are capable of stimulating IL 2 release and proliferative responses from other T cell clones. Evidence is presented suggesting that B lymphoma cell stimulation of these T cell clones is largely IL 1 independent and that at least some T cell clones may require activation signals other than Ia, antigen, and IL 1. The addition of exogenous, purified IL 1 to the T cell activation assays was found to have a wide range of stimulatory effects on the proliferative responses of different T cell clones. The absence of comparable IL 1-induced stimulation of IL 2 secretion suggests that IL 1 primarily enhances antigen specific T cell proliferation through mechanisms other than acting as a co-stimulant for IL 2 release. 相似文献
18.
Therapeutic implications of serum factors inhibiting proliferation and inducing differentiation of myeloid leukemic cells 总被引:2,自引:0,他引:2
Murine post-endotoxin sera contain high levels of myeloid colony-stimulating factor(s) (GM-CSF) and factors capable of inducing terminal granulocyte and macrophage differentiation of the murine myelomonocytic leukemic cell line WEHI-3. The combination of C. parvum and endotoxin induced a serum activity capable of inducing tumor necrosis and inhibiting leukemic colony formation in vitro. This factor (TNF) could be separated from the differentiation-inducing factor (GM-DF) and from CSF. In conjunction with a Phase I trial of highly purified endotoxin in patients with advanced malignancy, we monitored human post-endotoxin sera for CSF and GM-DF. Induction of GM-DF occurred maximally at 2-6 h and was associated with increased serum levels of CSF active against the patient's own bone marrow. Following repeated injections of escalating doses of endotoxin, persistent levels of GM-DF were detected both pre-endotoxin and 24 h post-endotoxin treatment. The ability to induce repeatedly a serum protein with potent capacity to promote terminal differentiation of myelomonocytic leukemic cells suggests a possible therapeutic role in human myeloid leukemias. 相似文献
19.
Culture media conditioned by several hepatocyte derived cell lines were analyzed for their ability to stimulate adipose differentiation of the adipogenic cell line 1246. The results presented here show that culture media from HepG2 and Hep3B cell lines contain a high level of the activity, whereas media from hepatoma and hepato adenocarcinoma cell lines Huh-7, PLC/PRF/5, and SK-Hep-1 do not contain adipogenic activity. Conditioned medium from HepG2 cells also stimulated differentiation of 3T3-L1 cells and of rat epididymal adipocyte precursors in primary culture. Partial biochemical characterization of the adipogenic activity carried out using HepG2 conditioned medium indicates that the hepatocyte derived adipogenic factor has an apparent molecular weight between 445 and 232 kDa, is destroyed by treatment at 100 degrees C, with protease, with 2-mercaptoethanol and in acidic conditions. The activity is stable at alkaline pH. Culture media conditioned by normal rat hepatocytes in primary culture also contained adipogenic activity. In contrast, medium conditioned by primary culture of nonhepatocyte cell also isolated from liver was deprived of this activity. The data presented in this paper suggest that hepatocytes could be a physiological site of production of adipogenic activity. 相似文献
20.
Belladonna ML Renauld JC Bianchi R Vacca C Fallarino F Orabona C Fioretti MC Grohmann U Puccetti P 《Journal of immunology (Baltimore, Md. : 1950)》2002,168(11):5448-5454
IL-23 is a recently discovered heterodimeric cytokine that shares biological properties with proinflammatory cytokines. The biologically active heterodimer consists of p19 and the p40 subunit of IL-12. IL-23 has been shown to possess biological activities on T cells that are similar as well distinct from those of IL-12. We have constructed single-chain IL-23 and IL-12 fusion proteins (IL-23-Ig and IL-12-Ig) and have compared the two recombinant proteins for effects on murine dendritic cells (DC). Here we show that the IL-23-Ig can bind a significant proportion of splenic DC of both the CD8alpha(-) and CD8alpha(+) subtypes. Furthermore, IL-23and IL-12-Ig exert biological activities on DC that are only in part overlapping. While both proteins induce IL-12 production from DC, only IL-23-Ig can act directly on CD8alpha(+) DC to promote immunogenic presentation of an otherwise tolerogenic tumor peptide. In addition, the in vitro effects of IL-23-Ig did not appear to require IL-12Rbeta2 or to be mediated by the production of IL-12. These data may establish IL-23 as a novel cytokine with major effects on APC. 相似文献