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B-myb alters the response of myeloid precursor cells to G-CSF

EXPERIMENTAL CELL RESEARCH 2000年第1期254卷 153-162页

作者： Engelhard, A Campbell, K Calabretta, B Thomas Jefferson Univ Kimmel Canc Inst Dept Microbiol & Immunol Philadelphia PA 19107 USA

The human B-myb gene encodes a cell cycle-regulated DNA-binding phosphoprotein which functions as a transcription factor with an important role in cell cycle progression, survival, and differentiation. Recently, it has been demonstrated that ectopic murine B-myb expression blocked the ability of 32Dc13 cells to proliferate in response to granulocyte colony-stimulating factor (G-CSF) and accelerated the induction of terminal differentiation. In contrast, we report that while 32Dc13 cells overexpressing human B-myb do display some markers of myeloid differentiation earlier than parental cells, including the expression of myeloperoxidase mRNA and the appearance of band myelocytes in G-CSF-induced cultures, the induction of late markers of differentiation is inhibited. The expression of lactoferrin mRNA is absent and the appearance of terminally differentiated polymorphonuclear cells is severely impaired in B-myb-expressing 32DcL3 cells. Furthermore, continuous exposure to G-CSF results in the outgrowth of a culture which expresses increased levels of B-myb RNA and is dependent on G-CSF for proliferation while retaining responsiveness to interleukin-3, These data suggest that the B-myb gene is involved in early transcriptional events during myeloid differentiation, but that its expression prevents terminal differentiation. (C) 2000 Academic Press.

关键词： CCAAT增强子结合蛋白类细胞周期蛋白质类细胞分化细胞分裂细胞系细胞存活 DNA结合蛋白质类/遗传学 DNA结合蛋白质类/代谢基因表达粒细胞集落刺激因子/代谢粒细胞集落刺激因子/药理学造血干细胞/细胞学造血干细胞/药物作用造血干细胞/代谢表型原癌基因蛋白质类/生物合成原癌基因蛋白质c-bcl-2/生物合成反式激活因子类/生物合成反式激活因子类/遗传学反式激活因子类/代谢转录因子/生物合成动物人类小鼠

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Distribution of granulocyte colony-stimulating factor (G-CSF) and G-CSF-receptor mRNA and protein in the human fetus

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PEDIATRIC RESEARCH 1999年第3期46卷 333-338页

作者： Calhoun, DA Donnelly, WH Du, Y Dame, JB Li, Y Christensen, RD Univ Florida Coll Med Dept Pediat Div Neonatol Gainesville FL 32610 USA Univ Florida Coll Med Dept Pathol Div Anat Pathol Gainesville FL 32610 USA

Granulocyte colony-stimulating factor (G-CSF) promotes neutrophil proliferation, differentiation, and survival. It acts by binding to specific cell-surface receptors (G-CSF-R), which are expressed on cells of granulocytic lineage, human endothelial cells, and placenta. It has been postulated that the administration of recombinant G-CSF (rG-CSF) to preterm neonates might be useful in treating infections or in reducing nosocomial infections. Whereas it is known that G-CSF and G-CSF-R are present in the developing fetal bone marrow and liver, no information is available as to the existence or distribution of nonhematopoietic G-CSF-R in other tissues of the developing human fetus. We hypothesized that G-CSF and its receptor might be expressed in various fetal tissues, as has been shown for other growth factors such as erythropoietin and fibroblast growth factor. Therefore, we studied the anatomical distribution of mRNA-encoding G-CSF and G-CSF-R, using RT-PCR and in situ RT-PCR in a variety of human fetal tissues ranging from 8 to 24 weeks postconception. The cellular distributions of the corresponding proteins were determined by immunohistochemistry. Both G-CSF and G-CSF-R were present in nearly every organ and tissue examined, but in discrete cellular localizations. G-CSF-R in kidney and intestine underwent changes in anatomical distribution with fetal development. These results indicate that G-CSF and G-CSF-R have wide anatomical expression in the developing human fetus.

关键词：胎儿/代谢粒细胞集落刺激因子/代谢免疫组织化学器官特异性聚合酶链反应 RNA 信使/分析 RNA 信使/生物合成受体粒细胞集落刺激因子/代谢人类

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Myeloblastin is a granulocyte colony-stimulating factor-responsive gene conferring factor-independent growth to hematopoietic cells

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 2000年第4期97卷 1601-1606页

作者： Lutz, PG Moog-Lutz, C Coumau-Gatbois, E Kobari, L Di Gioia, Y Cayre, YE Hop St Antoine INSERM U417 F-75012 Paris France Thomas Jefferson Univ Kimmel Canc Inst Dept Microbiol Immunol Philadelphia PA 19107 USA

Hematopoiesis depends on a pool of quiescent hematopoietic stem/progenitor cells. When exposed to specific cytokines, a portion of these cells enters the cell cycle to generate an amplified progeny. Myeloblastin (MBN) initially was described as involved in proliferation of human leukemia cells. The granulocyte colony-stimulating factor (G-CSF), which stimulates the proliferation of granulocytic precursors, up-regulates MBN expression. Here we show that constitutive overexpression of MBN confers factor-independent growth to murine bone marrow-derived Ba/F3/G-CSFR cells. Our results point to MBN as a C-CSF responsive gene critical to factor-independent growth and indicate that expression of the G-CSF receptor is a prerequisite to this process. A 91-bp MBN promoter region containing PU.1, C/EBP, and c-Myb binding sites is responsive to G-CSF treatment. Although PU.1. C/EBP, and c-Myb transcription factors all were critical for expression of MEN, its up-regulation by G-CSF was associated mainly with PU.1. These findings suggest that MBN is an important target of PU.1 and a key protease for factor-independent growth of hematopoietic cells.

关键词：抗原 CD34/免疫学 CCAAT增强子结合蛋白类 DNA结合蛋白质类/代谢基因表达调控粒细胞集落刺激因子/药理学造血干细胞/药物作用成髓细胞素核蛋白质类/代谢启动区遗传原癌基因蛋白质类/代谢原癌基因蛋白质c-myb/代谢受体粒细胞集落刺激因子/代谢丝氨酸内肽酶类/遗传学丝氨酸内肽酶类/代谢反式激活因子类/代谢转录因子/代谢转染肿瘤细胞培养的动物人类小鼠

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Reassessment of interactions between hematopoietic receptors using common beta-chain and interleukin-3-specific receptor beta-chain-null cells: no evidence of functional interactions with receptors for erythropoietin, granulocyte colony-stimulating factor, or stem cell factor

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BLOOD 2000年第4期96卷 1588-1590页

作者： Scott, CL Robb, L Papaevangeliou, B Mansfield, R Nicola, NA Begley, CG Walter & Eliza Hall Inst Med Res Melbourne Vic 3050 Australia Cooperat Res Ctr Cellular Growth Factors Melbourne Vic Australia Rotary Bone Marrow Res Labs Factors Melbourne Vic Australia

Mice lacking both the gene encoding the shared receptor for granulocyte macrophage-colony-stimulating factor (GMCSF), interleukin-3 (IL-3), and IL-5 common beta-chain (B-c) and the gene for the IL-3 specific receptor (BIL3) were generated. This was achieved by targeting the B-c locus in embryonic stem cells that were heterozygous for a null mutation of BIL3. Cells from mice generated with the doubly targeted embryonic stem cells were unresponsive to all 3 cytokines. Considerable previous data suggested a role for common beta-chain (beta(c)) in modulating signaling of cytokines including erythropoietin (EPO), G-CSF, and stem cell factor (SCF). However, bone marrow cells from mice lacking beta(c) and beta(IL3) showed normal responsiveness to these cytokines. Thus, there was no evidence for a biologically significant interaction between signaling via beta(c) or beta(IL3) and signaling by EPO, G-CSF, or SCF. Previously documented biochemical phenomena, including receptor transmodulation, receptor transphosphorylation, and even direct physical interaction, involving the beta(c)/beta IL-3 receptor systems do not reflect genuine interactions of physiological significance in primary hematopoietic cells. This study provided results that challenge conclusions previously established using a variety of biochemical assays. (Blood. 2000;96:1588-1590) (C) 2000 by The American Society of Hematology.

关键词：粒细胞集落刺激因子/代谢血细胞生成/遗传学白细胞介素3/代谢原癌基因蛋白质c-kit/遗传学原癌基因蛋白质c-kit/代谢受体交叉感知受体红细胞生成素/遗传学受体红细胞生成素/代谢受体粒细胞集落刺激因子/遗传学受体粒细胞集落刺激因子/代谢受体白细胞介素3/遗传学受体白细胞介素3/代谢序列缺失信号传导/遗传学干细胞因子/代谢动物小鼠

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Purification, crystallization and preliminary X-ray analysis of a complex between granulocyte colony-stimulating factor and its soluble receptor

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ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY 2000年第6期56卷 751-753页

作者： Aritomi, M Kunishima, N Okitsu, N Shimizu, M Ota, Y Morikawa, K Biomol Engn Res Inst Suita Osaka 5650842 Japan

Crystals of the complex between granulocyte colony-stimulating factor and its soluble receptor were obtained by a vapour-diffusion method using ammonium sulfate as a precipitant. Addition of 1,4-dioxane was critical in order to grow the crystals to sufficient sizes. Cryoprotection was essential in order to collect diffraction data at atomic resolution. Two kinds of crystal forms were obtained depending on the cryoprotectants. In a cryosolvent with the same salt concentration as in the crystallization conditions, the crystal belonged to the space group I4(1)22. At higher salt concentrations, the crystal was converted to a different space group P4(1)2(1)2 (P4(3)2(1)2) with the same unit-cell parameters.

关键词：碳水化合物构象结晶结晶学 X线/方法数据说明统计冷冻粒细胞集落刺激因子/化学粒细胞集落刺激因子/分离和提纯粒细胞集落刺激因子/代谢大分子物质肽碎片/化学肽碎片/分离和提纯蛋白质结构三级受体粒细胞集落刺激因子/化学受体粒细胞集落刺激因子/分离和提纯重组蛋白质类溶解度超速离心法动物人类小鼠

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Concentrations of granulocyte colony-stimulating factor in human milk after in vitro simulations of digestion

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PEDIATRIC RESEARCH 1999年第6期46卷 767-771页

作者： Calhoun, DA Lunoe, M Du, Y Staba, SL Christensen, RD Univ Florida Div Neonatol Dept Pediat Coll Med Gainesville FL 32610 USA

Human milk contains proteins that survive digestion in the neonatal gastrointestinal tract. Our group and others have reported that granulocyte colony-stimulating factor (G-CSF), a hematopoietic cytokine that influences neutrophil proliferation and differentiation, is present in human milk. We also reported that specific receptors for G-CSF are expressed on the villous enterocytes of neonates. However, the physiologic role of milk-borne G-CSF is not known. Thus, we sought to evaluate the capacity of human milk to protect G-CSF against proteolytic degradation after exposure to gastric secretions obtained from preterm (PT) and term (T) neonates at pH concentrations of 3.2, 5.8, and 7.4. Specifically, we examined degradation of 1) endogenous G-CSF in PT (n = 15) and T (n = 15) human milk;2) recombinant human G-CSF (rhG-CSF) added to a protein-free buffer (n = 10, 5 PT and 5 T);and 3) rhG-CSF added to human milk (n = 12, 6 PT and 6 T), various commercially prepared infant formulas tit = 15), and cow's milli (n = 5). Endogenous G-CSF and rhG-CSF added to human milk resisted degradation at 1 and 2 h. However, when rhG-CSF was added to commercial formulas, >95% was degraded at 1 and 2 h at each pH level. Similarly, approximately 60% of rhG-CSF added to cow's milk was degraded at 1 and 2 h. We conclude that I) endogenous G-CSF and rhG-CSF added to human milk are protected from degradation after exposure to gastric secretions at physiologic pH levels, 2) rhG-CSF added to infant formulas is not protected from degradation, and 3) it is likely that the G-CSF present in human milk is biologically available to the neonate.

关键词：消化粒细胞集落刺激因子重组/药理学粒细胞集落刺激因子/代谢乳人/代谢人类

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