CONTROL OF ERYTHROPOIESIS BY GROWTH PROMOTING PEPTIDES

多种多肽生长因子(GFS)表达红系 在培养中的活性，也可能在体内的活性。最近的 用于哺乳动物细胞的化学定义培养液的可用性 培养允许评估激素的影响和调节 在细胞和亚细胞水平上的机制。我们将聘请一名 用于培养人红系细胞的血清“游离”系统 我们的发展是为了： 1.确定已定义的GFS的细胞效应。我们会 确定选定多肽的剂量-反应关系 在去血清的单层和双层培养中。 我们将扩大我们的发现，一位GF，L-三碘甲腺原氨酸， 不仅刺激蛋白质释放，还刺激旁分泌(猝发 促进活性，双酚A)从培养的 淋巴细胞。蛋白质和双酚A的累积释放率 放射性标记生长因子Will的释放及亚细胞定位 被定义。特定的细胞内处理路径将 被研究。新合成的双酚A类化合物的相互关系 未诱导的分子和有丝分裂原驱动的双酚A 在生物化学和免疫学上有定义。 2.分离提纯未定义的GFS。可溶性双酚A随药物释放 在没有诱导剂的情况下可以从液体中提纯 5~6成熟。柱状计时，凝集素亲和力 色谱和反相高效液体- 将使用层析法。此外，抗双酚A免疫球蛋白将 用于免疫亲和层析纯化。 3.鉴定成纤维细胞生长因子靶细胞并检测成纤维细胞生长因子结合。我们 将利用杂交瘤抗体选择性地为 红系祖细胞。与抗T-T-单抗结合 和B细胞群，它们将被用来识别骨髓 对生长因子效应敏感或抗性的亚群。 胰岛素和胰岛素样生长因子与PANED结合 骨髓细胞也将被确定。 4.体内反应和组织培养反应的相关性。我们会 对低增殖性贫血患者进行检测 体外雄激素敏感性试验的预测价值。 慢性粒细胞白血病患者血浆和骨髓细胞的研究 增生性骨髓营养不良将发掘潜力 涉及血小板衍生性生长的体液异常 因子(PDGF)。这些调查着眼于我们的长期 评估组织的生理相关性的目标 健康和疾病方面的培养结果。

Many polypeptide growth factors (GFs) express erythropoietic activity in culture and possibly in vivo as well. The recent availability of chemically defined media for mammalian cell culture permits evaluation of hormone effects and regulatory mechanisms at cellular and subcellular levels. We will employ a serum "free" system for the culture of human erythroid cells that we have developed in order to: 1. Determine cellular effects of defined GFs. We will determine dose-response relationships for selected peptides in single and double layer cultures under serum deprivation. We will extend our finding that one GF, L-triiodothyronine, stimulates not only protein release but also paracrine (burst promoting activity, BPA) release from cultured lymphocytes. Cumulative protein and BPA release, rates of release and subcellular localization of radiolabeled GF will be defined. Specific intracellular processing pathways will be studied. The relationship of newly synthesized BPA-like molecules to uninduced and mitogen-driven BPA will be defined biochemically and immunologically. 2. Isolate and purify undefined GFs. Soluble BPA released with and without inducting agents will be purified from liquid medium. Column chronmatograpy, lectin affinity chromatograpy and reverse-phase high-performance liquid- chromatography will be used. In addition, anti-BPA IgG will be used for purification by immunoaffinity chromatography. 3. Characterize GF target cells and examine GF binding. We will utilize hybridoma antibodies to selectively "pan" for erythroid progenitors. Together with monoclonal IgG to T- and B-cell populations, they will be used to identify marrow subpopulations that are sensitive or resistant to GF effects. Insulin and insulin-like growth factor binding to panned marrow cells will also be determined. 4. Correlate in vivo and tissue culture responses. We will accure patients with hypoproliferative anemia to determine the predictive value of in vitro androgen sensitivity testing. Studies of plasma and marrow cells from patients with hyperproliferative marrow dyscrasias will explore potential humoral abnormalities involving platelet-derived growth factor (PDGF). These investigations focus on our long term goal of evaluating the physiological relevance of tissue culture findings in health and disease.