LOW DOSE RATE IRRADIATION EFFECT ON HEMOPOIESIS IN VITRO

低剂量率辐照对体外造血作用的影响

• 批准号: 3181103
• 负责人: JOEL S GREENBERGER
• 金额: $ 15.03万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-08-01 至 1988-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3181103
• 关键词: X ray; cell population study; colony stimulating factor; gamma radiation; hematopoiesis; hematopoietic stem cells; ionizing radiation; megakaryocytes; radiation dosage; relative biological effectiveness; tissue /cell culture

Both high and low dose rate x-irradiation of purified marrow stromal cells in vitro have been demonstrated by us to alter the growth of added purified hematopoietic stem cells. Dose rates of 5 or 20 rad/min (low) stabilize some parameters of hematopoiesis significantly better than an equivalent leukemia-cell lethal standardized dose delivered at 200 rad min (high), including: production and release from stromal cells of growth factors including granulocyte-macrophage colony stimulating factor-1 (FM-CSF-1) required for proliferation in agar overlay of committed granulocyte-macrophage progenitor cells (GM-CFUc). However, other parameters of hematopoiesis including cumulative production in liquid overlay of total hemopoietic cells and pluripotential hematopoietic stem cells (CFUs) are equally depressed at both low and high dose rates. These physiologic alterations of stromal cells are independent of proliferative integrity as measured by clonagenic survival of replated cells. The protective effect of low dose rate irradiation is lost by prior in vitro exposure of stromal cells to the chemotherapeutic alkylating agent, L-PAM. We will now determine the mechanism of these non-lethal physiologic effects of irradiation on marrow stromal cells, and the critical dose rate for each detectable biologic effect. Methods will involve continuous bone marrow cultures; preparation of purified stromal and hematopoietic stem cell cultures; identification of the phenotype of stromal cells surviving each irradiation dose; measurement of self-renewal capacity of CFUs; quantitation of levels of synthesis of multipotential cell CSF also known as Interleukin-3 (IL-3) and CSF-1; and study of the kinetics of repair of x-ray damage in these compared to other stromal cell cultures that have been treated prior to irradiation with the aklylating agent, L-Phenylalanine mustard; nitrosyl urea; adriamycin or busulfan in vitro. These studies should: improve understanding of irradiation effects that alter cell to cell interaction; and aid in modification of the sequence of chemotherapy and total body irradiation in drug/x-ray protocols to optimize clinical marrow engraftment and improve survival in bone marrow transplantion.

高、低剂量率X射线照射对纯化骨髓基质细胞的影响 我们在体外已经证明了改变添加的纯化的生长 造血干细胞。5或20rad/min(低)的剂量率稳定 某些造血参数明显优于等效值 白血病细胞致死标准剂量在200拉德分(高)， 包括：基质细胞产生和释放生长因子 包括粒细胞巨噬细胞集落刺激因子-1(FM-CSF-1) 在已承诺的琼脂覆盖层中增殖所需的 粒细胞-巨噬细胞前体细胞(GM-CFUc)。然而，其他 包括液体累积产生量在内的造血参数 总造血细胞和多潜能造血干细胞的重叠 细胞(CFU)在低剂量率和高剂量率下都同样受到抑制。这些 基质细胞的生理性改变与增殖无关 通过复制细胞的克隆形成存活来衡量完整性。这个 低剂量率照射对小剂量辐射的保护作用 将间质细胞暴露在化疗烷化剂L-PAM中。 我们现在将确定这些非致命性生理效应的机制 对骨髓基质细胞的照射剂量，以及每种细胞的临界剂量率 可检测的生物效应。方法将涉及持续骨髓 培养；纯化基质干细胞和造血干细胞的制备 培养物.存活的基质细胞表型的鉴定 辐射剂量；CFU自我更新能力的测量； 多潜能细胞脑脊液合成水平的定量也已知 作为白细胞介素3(IL-3)和脑脊液-1(CSF-1)；修复动力学的研究 与其他基质细胞培养相比，这些细胞的X射线损伤 在辐照前用烷化剂处理， L-苯丙氨酸芥末；亚硝酰脲；阿霉素或丁硫丹。 这些研究应该：提高对辐射影响的理解， 改变细胞与细胞之间的相互作用；以及辅助修改 化疗和全身照射在药物/X射线方案中的优化 临床骨髓移植与提高骨髓存活率 移植手术。

项目成果

Enhancer sequences of a retroviral vector determine expression of a gene in multipotent hematopoietic progenitors and committed erythroid cells.

逆转录病毒载体的增强子序列决定多能造血祖细胞和定向红系细胞中基因的表达。

• DOI: 10.1073/pnas.84.23.8662
• 发表时间: 1987
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Holland,CA;Anklesaria,P;Sakakeeny,MA;Greenberger,JS
• 通讯作者: Greenberger,JS

Expression of transfected recombinant oncogenes increases radiation resistance of clonal hematopoietic and fibroblast cell lines selectively at clinical low dose rate.

转染的重组癌基因的表达在临床低剂量率下选择性地增加克隆造血细胞和成纤维细胞系的辐射耐受性。

• 发表时间: 1990
• 期刊: Radiation research
• 影响因子: 3.4
• 作者: FitzGerald,TJ;Henault,S;Sakakeeny,M;Santucci,MA;Pierce,JH;Anklesaria,P;Kase,K;Das,I;Greenberger,JS
• 通讯作者: Greenberger,JS

Infection of hematopoietic and stromal cells in human continuous bone marrow cultures by a retroviral vector containing the neomycin resistance gene.

通过含有新霉素抗性基因的逆转录病毒载体感染人连续骨髓培养物中的造血细胞和基质细胞。

• DOI: 10.1159/000205362
• 发表时间: 1989
• 期刊: Acta haematologica
• 影响因子: 2.4
• 作者: Holland,CA;Rothstein,L;Sakakeeny,MA;Anklesaria,P;Griffin,JD;Harigaya,K;Newburger,PE;Greenberger,JS
• 通讯作者: Greenberger,JS

Biological characterization of cloned permanent stromal cell lines from anemic Sl/Sld mice and +/+ littermates.

贫血 Sl/Sld 小鼠和/同窝小鼠克隆永久基质细胞系的生物学特征。

• 发表时间: 1987
• 期刊: Experimental hematology
• 影响因子: 2.6
• 作者: Anklesaria,P;Klassen,V;Sakakeeny,MA;FitzGerald,TJ;Harrison,D;Rybak,ME;Greenberger,JS
• 通讯作者: Greenberger,JS

Homing of a cloned multipotential stem cell line in spleen and intraperitoneal membrane.

克隆的多能干细胞系在脾脏和腹膜内膜中的归巢。

• 发表时间: 1986
• 期刊: Experimental hematology
• 影响因子: 2.6
• 作者: Hardy,CL;Kishimoto,T;Harjes,K;Tavassoli,M;Greenberger,JS
• 通讯作者: Greenberger,JS