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DIFFERENTIATION OF POPULATION OF PERIPHERAL-BLOOD LYMPHOCYTES INTO CELLS BEARING SHEEP ERYTHROCYTE RECEPTORS INVITRO BY HUMAN THYMIC EXTRACT

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 1975年第3期72卷 1175-1178页

作者： VOGEL, JE INCEFY, GS GOOD, RA MEM SLOAN KETTERING CANC CTR LAB CLIN IMMUNOBIOL 1275 YORK AVE NEW YORK NY 10021 USA

A small population of human marrow cells has been shown to be differentiated in vitro by thymic extract into cells bearing T-lymphocyte (thymus-derived lymphocyte) characteristics. By a similar method, the differentiation of human peripheral blood lymphocytes has been studied. A discontinuous gradient of bovine serum albumin was used to isolate lymphocytes into four layers and cells from layers I and III demonstrated the greatest potential for differentiation by human thymic extract. Appearance of T-lymphocyte characteristics was recognized by the spontaneous E-rosette technique with sheep erythrocytes. Ability of human marrow cells to be differentiated under the influence of human thymic extract was abolished by specific inhibitors of nucleic acid synthesis, however, had no inhibitory effect on the maturation of peripheral blood lymphocytes during a 2 hr incubation with human thymic extract but puromycin, an inhibitor of protein synthesis, abolished this differentiative step in cells of layer I. It is suggested from these studies that many of the cells in peripheral blood that are differentiable by thymic extract are at a stage of maturation more advanced than those in human marrow that are also differentiable by thymic extract.

关键词：鹅膏菌素类/药理学细胞分化/药物作用细胞膜/免疫学细胞分离离心法梯密度更生霉素/药理学红细胞/免疫学造血干细胞/细胞学造血干细胞/药物作用免疫黏连反应 RNA/生物合成绵羊/免疫学 T淋巴细胞/细胞学 T淋巴细胞/药物作用 T淋巴细胞/代谢胸腺提取物/药理学动物人类

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

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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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Mouse mammary tumor virus superantigen expression is reduced by glucocorticoid treatment

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VIROLOGY 2000年第1期275卷 98-106页

作者： Sepúlveda, VAT Berdel, B Coffin, JM Reuss, FU Deutsch Krebsforschungszentrum Forschungsschwerpunkt Angew Tumorvirol F0400 D-69120 Heidelberg Germany Tufts Univ Sch Med Dept Mol Biol & Microbiol Boston MA 02111 USA

Expression of mouse mammary tumor virus (MMTV)-encoded superantigens in B lymphocytes are required for viral transmission and pathogenesis. Due to problems with detection and quantification of the superantigen protein, most reports about the mechanism of superantigen expression from the viral sag gene rely on the quantitative analysis of putative sag mRNAs. The description of multiple promoters as a source of putative sag mRNA has complicated the situation even further. All conclusions about the level of superantigen protein expression based on these data remain circumstantial. To test the effect of the glucocorticoid hormone dexamethasone on the total superantigen expression from an infectious MMTV provirus we used a quantitative assay that is based on a superantigen-luciferasa fusion protein. MMTV gene expression from the major promoter in the 5' long terminal repeat (LTR) is strongly induced in the presence of glucocorticoid hormones. We now demonstrate that, in the presence of dexamethasone, sag gene expression is reduced despite increased transcription from the MMTV 5' LTR and increased amounts of putative sag mRNA initiated at the LTR promoter. These data show that the expression of the MMTV sag gene does not correlate with the activity of the major LTR promoter and thus differs from all Other MMTV genes. (C) 2000 Academic Press.

关键词：细胞系地塞米松/药理学地塞米松/治疗应用基因表达调控病毒/药物作用基因报告/遗传学基因病毒/遗传学糖皮质激素类/药理学糖皮质激素类/治疗应用造血干细胞/药物作用造血干细胞/代谢造血干细胞/病毒学乳房瘤病毒鼠/药物作用乳房瘤病毒鼠/遗传学乳房瘤病毒鼠/免疫学启动区(遗传学)/遗传学原病毒/药物作用原病毒/遗传学原病毒/免疫学 RNA 信使/分析 RNA 信使/遗传学 RNA 病毒/分析 RNA 病毒/遗传学重组融合蛋白质类/遗传学超抗原/遗传学末端重复序列/遗传学转录遗传/药物作用转录遗传/遗传学转染动物小鼠

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Recombinant human interleukin-11 synergizes with steel factor and interleukin-3 to promote directly the early stages of murine megakaryocyte development in vitro

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BLOOD 2000年第2期95卷 503-509页

作者： Weich, NS Fitzgerald, M Wang, AL Calvetti, J Yetz-Aldape, J Neben, S Turner, KJ Genet Inst Dept Tissue Growth & Repair Cambridge MA 02140 USA Genet Inst Dept Immunol Cambridge MA 02140 USA

The authors studied the role that interleukin (IL)-11 plays during the early stages of megakaryocyte (MK) development by investigating its in vitro effects on cell subpopulations enriched for bone marrow primitive progenitor cells and early and late committed progenitor cells. Progenitor subpopulations were isolated from bone marrow of normal or 5-fluorouracil (5FU)-treated mice and separated by sorting based on the surface antigens Sca-1, c-kit, and CD34. Functional analysis of the cell subpopulations, 5FU Lin(-)Sca-1(+)c-kit(+) or normal bone marrow (NBM) Lin(-)Sca-1(+)c-kit(+)CD34(-)cells, indicated that exposure of these cells to recombinant human (rh)IL-11 in combination with steel factor (SF) stimulates the formation of colonies in methylcellulose and their proliferation in single cell-containing liquid cultures. Kinetic studies of MK progenitor generation, in response to SF and rhIL-11, demonstrated that a significant number of the progenitors produced are committed to the MK lineage. RhlL-11 also synergized with both SF and IL-3 to stimulate MK colony growth from NBM Lin(-)Sca-1(+)c-kit(+) cells (early progenitors) and NBM Lin(-)Sca-1(-)c-kit(+) cells (committed late progenitors). In the presence of IL-3, NBM, Lin(-)Sca-1(-)c-kit(+) cells responded more strongly to rhIL-11 than SF. Consistent with these results is the observation that IL-11 receptor alpha chain mRNA is present in all the progenitor cells from which the MKs are derived. This cell culture and RNA analysis suggest that murine bone marrow primitive progenitor cells and early and late progenitor cells are direct targets of rhIL-11 and that rhIL-11 has the potential to promote megakaryocyte development at several very early stages. (Blood, 2000;95:503-509) (C) 2000 by The American Society of Hematology.

关键词：抗原 CD34/分析抗原 Ly/分析骨髓细胞/细胞学细胞分化/药物作用细胞分离/方法细胞培养的药物协同作用基因表达调控/免疫学造血干细胞/细胞学造血干细胞/药物作用白细胞介素11/药理学白细胞介素11受体α亚单位白细胞介素3/药理学巨核细胞/细胞学巨核细胞/药物作用膜蛋白质类/分析小鼠近交C57BL 原癌基因蛋白质c-kit/分析受体白细胞介素/遗传学受体白细胞介素11 重组蛋白质类/药理学干细胞因子/药理学动物女(雌)性人类小鼠

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Cell death and cell-cycle arrest induced by incorporation of [³H]thymidine into human haemopoietic cell lines

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INTERNATIONAL JOURNAL OF RADIATION BIOLOGY 2000年第3期76卷 295-303页

作者： Yanokura, M Takase, K Yamamoto, K Teraoka, H Tokyo Med & Dent Univ Med Res Inst Chiyoda Ku Tokyo 1010062 Japan Tokyo Med & Dent Univ Dept Med Informat Bunkyo Ku Tokyo 1138510 Japan

Purpose: To investigate the influence of [methyl-H-3]thymidine ([H-3]Tdr) incorporated into human haemopoietic cell lines. Materials and methods: HL-60, Molt-4, Jurkat, Raji and SRW6-GCL4 cells were incubated in the presence of [H-3]Tdr. Cell proliferation, cell viability, DNA fragmentation and expression of caspase-3 and Bcl-2 families were examined. The cell-cycle of HL-60 was analysed using flow cytometry. Results: In HL-60, Molt-4 and Jurkat, cell death was accompanied by DNA nucleosomal fragmentation and activation of caspase-3. In Raji and SKW6-CL4, it was accompanied by neither. Protein levels of Bcl-2 and Bad in HL-60 and Molt-4 did not significantly change, and that of Bax was decreased after a 3-day incubation. HL-60 incubated in the presence of 74 or 185 kBq/ml [H-3]Tdr arrested at G(2)/M phase, and then underwent apoptosis. In 7.4kBq/ml, the cell-cycle progressed after the delay in S-phase. Conclusions: Two different modes of cell death were observed when [H-3]Tdr was incorporated into the human haemopoietic cell lines. Incorporation into HL-60 cells resulted in delay of S-phase progression, arrest at G(2)/M and apoptosis.

关键词：印迹法蛋白质钙结合蛋白质类/代谢载体蛋白质类/生物合成半胱氨酸天冬氨酸蛋白酶1/拮抗剂和抑制剂半胱氨酸天冬氨酸蛋白酶1/代谢半胱氨酸天冬氨酸蛋白酶3 半胱氨酸天冬氨酸蛋白酶/拮抗剂和抑制剂半胱氨酸天冬氨酸蛋白酶/代谢细胞周期/药物作用细胞周期/辐射效应细胞死亡/药物作用细胞死亡/辐射效应细胞分裂/药物作用细胞分裂/辐射效应细胞系细胞存活/药物作用细胞存活/辐射效应半胱氨酸蛋白酶抑制剂/药理学 DNA碎片裂/药物作用剂量效应关系辐射流式细胞术造血干细胞/药物作用造血干细胞/代谢造血干细胞/辐射效应寡肽类/药理学聚ADP核糖聚合酶类/代谢原癌基因蛋白质类/生物合成原癌基因蛋白质c-bcl-2/生物合成胸苷/药代动力学胸苷/药理学氚 bcl-2相关X蛋白质 bcl相关蛋白质人类

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BLOOD 2000年第12期95卷 3716-3724页

作者： Zamai, L Secchiero, P Pierpaoli, S Bassini, A Papa, S Alnemri, ES Guidotii, L Vitale, M Zauli, G Univ Urbino Inst Morphol Sci I-61029 Urbino Italy Univ Ferrara Dept Morphol & Embryol Human Anat Sect I-44100 Ferrara Italy Univ Bologna Inst Histol & Gen Embryol Bologna Italy Thomas Jefferson Univ Jefferson Med Coll Kimmel Canc Inst Philadelphia PA 19107 USA Univ Brescia Dept Biomed Sci & Biotechnol Human Anat Sect Brescia Italy CNR NP Inst Cytomorphol Codivilla Putti Res Inst I-40126 Bologna Italy Univ Chieti Inst Human Morphol G DAnnunzio Chieti Italy

The impact of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) on normal hematopoietic development was investigated using adult peripheral blood CD34(+) hematopoietic progenitor cells, induced to differentiate along the erythroid, megakaryocytic, granulocytic, and monocytic lineages by the addition of specific cytokine cocktails. TRAIL selectively reduced the number of erythroblasts, showing intermediate levels of glycophorin A (glycophorin A(interm)) surface expression, which appeared in liquid cultures supplemented with stem cell factor + interleukin 3 + erythropoietin at days 7-10. However, neither immature (day 4) glycophorin A(dim) erythroid cells nor mature (day 14) glycophorin A(bright) erythroblasts were sensitive to TRAIL-mediated apoptosis, Moreover, preexposure to TRAIL significantly decreased the number and size of erythroid colonies in semisolid assays, These adverse effects of TRAIL were selective for erythropoiesis, as TRAIL did not significantly influence the survival of cells differentiating along the megakaryocytic, granulocytic, or monocytic lineages, Furthermore, TRAIL was detected by Western blot analysis in lysates obtained from normal bone marrow mononuclear cells, These findings indicate that TRAIL acts in a lineage- and stage of differentiation-specific manner, as a negative regulator of normal erythropoiesis. (C) 2000 by The American Society of Hematology.

关键词：抗原 CD/分析抗原 CD15/分析抗原 CD34/分析细胞凋亡/药物作用凋亡调节蛋白质类骨髓细胞/细胞学细胞存活/药物作用细胞培养的集落形成单位测定红细胞生成/药物作用红细胞生成/生理学流式细胞术造血干细胞/细胞学造血干细胞/药物作用造血干细胞/生理学配体膜糖蛋白类/药理学重组蛋白质类/药理学 TNF相关凋亡诱导配体肿瘤坏死因子α/药理学成年人人类

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Hypersensitivity of circulating progenitor cells to megakaryocyte growth and development factor (PEG-rHu MGDF) in essential thrombocythemia

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BLOOD 2000年第10期96卷 3310-3321页

作者： Axelrad, AA Eskinazi, D Correa, PN Amato, D Univ Toronto Dept Anat & Cell Biol Toronto ON M5S 1A8 Canada Univ Toronto Dept Med Biophys Toronto ON M5S 1A8 Canada Mt Sinai Hosp Dept Med Hematol Oncol Toronto ON Canada

Hematopoietic progenitor cells in 2 myeloproliferative disorders, juvenile chronic myelomonocytic leukemia and polycythemia vera, are known to be hypersensitive to cytokines that control normal progenitor cell proliferation, differentiation, and survival in their respective granulocyte/macrophage and erythroid lineages. Because thrombopoietin controls these functions in the normal megakaryocytic lineage, we asked the question: Are megakaryocytic progenitor cells in the myeloproliferative disorder essential thrombocythemia (ET) hypersensitive to thrombopoietin? Peripheral blood mono-nuclear cells from patients with ET, or secondary (reactive) thrombocytosis (2 degreesT), or healthy volunteers were grown in strictly serum-free agarose culture containing interleukin 3 (IL-3) and all-transretinoic acid, with various concentrations of PEG-rHu megakaryocyte growth and development factor (MGDF), The concentration of cytokine at half-maximum colony number served as a measure of progenitor cell sensitivity. Hypersensitivity to PEG-rHu MGDF was found in circulating progenitors from 18 of 20 (90%) informative patients with presumptive diagnosis ET, 1 of 8 (12.5%)2 degreesT patients, and none of the 22 healthy volunteers. Median MGDF sensitivity ratio in ET patients was approximately 53 times greater than in the controls. This hypersensitivity, which was also directed to rHu thrombopoietin, was highly specific with respect to cytokine, disease, and cell lineage. We propose that, despite their single pluripotential cell origin, the different clinicopathologic phenotypes in different chronic myeloproliferative disorders are determined by lineage-restricted hypersensitivities of hematopoietic progenitor cells to endogenous cytokines, This work emphasizes the importance of stringent serum-free conditions for revealing true sensitivities to cytokines, The findings also offer a basis for evolving a positive test for ET, a diagnosis now made essentially by exclusion. (C) 2000 by The A

关键词：血液循环细胞培养技术/方法队列研究集落形成单位测定细胞因子类/药理学剂量效应关系药物药物过敏/病因学造血干细胞/药物作用巨核细胞/药物作用巨核细胞/病理学巨核细胞/超微结构聚乙烯二醇类/药理学重组蛋白质类/药理学血小板增多出血性/血液血小板增多出血性/病因学血小板生成素/药理学成年人老年人老年人 80以上女(雌)性人类男(雄)性中年人

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Identification of podocalyxin-like protein 1 as a novel cell surface marker for hemangioblasts in the murine aorta-gonad-mesonephros region

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IMMUNITY 1999年第5期11卷 567-578页

作者： Hara, T Nakano, Y Tanaka, M Tamura, K Sekiguchi, T Minehata, K Copeland, NG Jenkins, NA Okabe, M Kogo, H Mukouyama, Y Miyajima, A Univ Tokyo Inst Mol & Cellular Biosci Bunkyo Ku Tokyo 1130032 Japan Univ Tokyo Core Res Evolut Sci & Technol Bunkyo Ku Tokyo 1130032 Japan Tokyo Univ Pharm & Life Sci Hachioji Tokyo 1920355 Japan NCI Mammalian Genet Lab ABL Basic Res Program Frederick Canc Res & Dev Ctr Frederick MD 21702 USA Osaka Univ Genome Informat Res Ctr Suita Osaka 5650871 Japan

Recent studies with avian embryos and murine embryonic stem cells have suggested that hematopoietic cells are derived from hemangioblasts, the common precursors of hematopoietic and endothelial cells. We molecularly cloned podocalyxin-like protein 1 (PCLP1) as a novel surface marker for endothelial-like cells in the aorta-gonad-mesonephros (AGM) region of mouse embryos, where long-term repopulating hematopoietic stem cells (LTR-HSCs) are known to arise. PCLP1(+) CD45(-) cells in the AGM region incorporated acetylated low-density lipoprotein and produced both hematopoietic and endothelial cells when cocultured with OP9 stromal cells. Moreover, multiple lineages of hematopoietic cells were generated in vivo when PCLP1(+)CD45(-) cells were injected into neonatal liver of busulfan-treated mice. Thus, PCLP1 can be used to separate hemangioblasts that give rise to LTR-HSCs.

关键词：氨基酸序列抗原 CD45/分析主动脉/化学主动脉/胚胎学生物学标记细胞分化/药物作用细胞谱系细胞培养的染色体图共同培养技术内皮血管/细胞学内皮血管/胚胎学胎儿蛋白质类/分析孕龄性腺/化学性腺/胚胎学生长物质/药理学造血干细胞移植造血干细胞/化学造血干细胞/细胞学造血干细胞/药物作用脂蛋白类 LDL/代谢膜糖蛋白类/分析膜糖蛋白类/遗传学中肾/化学小鼠近交C57BL 小鼠转基因分子序列数据抑瘤素M 肽类/药理学肽类/生理学干细胞/化学干细胞/分类干细胞/药物作用动物小鼠

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Effects of in vivo administration of adamantylamide dipeptide on bone marrow granulocyte-macrophage hemopoietic progenitor cells (GM-CFC) and on ability of serum of the treated mice to stimulate GM-CFC colony formation in vitro:: Comparison with muramyl dipeptide and glucan

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IMMUNOPHARMACOLOGY AND IMMUNOTOXICOLOGY 1999年第1期21卷 1-14页

作者： Vacek, A Hofer, M Masek, K Acad Sci Czech Republic Inst Biophys CS-61265 Brno Czech Republic Acad Sci Czech Republic Inst Pharmacol Prague 14220 4 Czech Republic

Adamantylamide dipeptide (AdDP), muramyl dipeptide (MDP), and glucan were shown to increase significantly the numbers of granulocyte-macrophage hemopoietic progenitor cells (GM-CFC) in the bone marrow of mice. However, whereas the sera of mice given MDP or glucan were found to stimulate the growth of colonies from GM-CFC in vitro, i.e. to produce a colony-stimulating activity (CSA), administration of AdDP did not lead to this effect. Nevertheless, when serum of mice given AdDP was added to the cultures concomitantly with a suboptimal concentration of mouse interleukin-3 (mIL-3), a broad spectrum hemopoietic stimulator, counts of colonies from GM-CFC were significantly increased, and accelerated growth of the colonies was found as well. This property of AdDP, i.e. its ability to exhibit co-stimulating activity (CoSA) without being able to exhibit CSA, suggests that AdDP acts in hemopoietic tissues differently as compared with the two other immunomodulators studied. It can be hypothesized that the action of AdDP is more specific when compared with its natural related compound, MDP, as well as with glucan. Our findings prove the possibility td stimulate by AdDP the granulopoietic compartment of hemopoiesis and are in agreement with previous observations concerning the absence of systemic side effects of AdDP. Both these qualities of AdDP may be advantageous when pondering over contingent clinical utilization of AdDP as hemopoietic stimulator.

关键词：乙酰胞壁酰-丙氨酰-异谷酰胺/免疫学乙酰胞壁酰-丙氨酰-异谷酰胺/药理学佐剂免疫/药理学金刚烷胺/类似物和衍生物金刚烷胺/免疫学金刚烷胺/药理学骨髓细胞/药物作用骨髓细胞/免疫学细胞培养的交叉遗传二肽类/免疫学二肽类/药理学葡聚糖类/免疫学葡聚糖类/药理学粒细胞/药物作用粒细胞/免疫学造血干细胞/药物作用造血干细胞/免疫学免疫血清/免疫学巨噬细胞/药物作用巨噬细胞/免疫学小鼠近交C57BL 小鼠近交CBA 动物小鼠

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HEMATOLOGIC EFFECTS OF ALCOHOL

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ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 1975年第APR25期252卷 307-315页

作者： HERBERT, V TISMAN, G COLUMBIA UNIV COLL PHYS & SURG DEPT PATHOL NEW YORK NY 10032 USA VET ADM HOSP BRONX NY 10468 USA

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