MAST CELL DIFFERENTIATION IN VITRO

肥大细胞体外分化

• 批准号: 2003441
• 负责人: THOMAS F HUFF
• 金额: $ 19.73万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2003441
• 关键词: Nippostrongylus; RNase protection assay; animal genetic material tag; antibody receptor; antireceptor antibody; biological signal transduction; bone marrow; carboxypeptidase; cell differentiation; colony stimulating factor; embryonic stem cell; enzyme linked immunosorbent assay; gene expression; hematopoiesis; interleukin 3; laboratory mouse; laboratory rat; lymph nodes; mast cell; peritoneum; polymerase chain reaction; protooncogene; tissue /cell culture; transcription factor

Mast cells are the major effector cells in diseases of immediate hypersensitivity. Understanding how mast cells develop from primitive blood cells may have implication for treatment of certain allergic diseases. Because mast cells may be uniquely related to hematopoietic stem cells, studies of early events in mast cell differentiation may have broader implications for hematopoiesis in general. The research proposed in this grant application extends the mast cell model system developed in the previous grant period. This system takes advantage of the fact that committed progenitors, but not uncommitted ones, can respond to stem cell factor alone by production of mast cell colonies in methylcellulose. There is continuing attention to the different roles that the c-kit/stem cell factor receptor-ligand complex may play in different stages of differentiation may have broader implications for hematopoiesis in general. The research proposed in this grant application extends the mast cell model system developed in the previous grant period. This system takes advantage of the fact that committed progenitors, but not uncommitted ones, can respond to stem cell factor alone by production of mast cell colonies in methylcellulose. There is continuing attention to the different roles that the c-kit/stem cell factor receptor-ligand complex may play in different stages of differentiation; however, the major focus in this grant application addresses Fc receptors on mast cells and their progenitors, because of recent provocative findings that in mast cells, both FcRI and FcRIII associated gamma chain dimers to transduce signals for activation and cytokine biosynthesis. We are particularly interested in the possibility that an induced autocrine IL-3 loop may be critical element in the committed state of these progenitors. The first specific aim is to determine whether bone marrow progenitors, particularly those which may be part of the early mast cell lineage, bear FcRI and FcRIII and if these receptors transduce a signal to induce an autocrine IL-3 loop which is necessary for commitment. In the second specific aim, mast cell-committed progenitors or granulated mast cells from MLN or peritoneal washouts will be tested to determine if FcRI/FcRIII-mediated signalling affects expression of autocrine cytokines, GATA transcription factors, or proteases. We will also determine if the continued occupancy of FcRI on mast cells in necessary for continued division of successive generations of daughter cells. The third specific aims takes advantage of an exciting new models system for the study of hematopoiesis; embryonic stem (ES) cell cultures. Once these very primitive cells are allowed to differentiate in culture, the expression of message and protein for FcRI/FcRIII will be monitored to correlate with the appearance of mast cells and autocrine IL-3 inducibility. These experiments may reveal similarities or differences between proposed in this grant application should contribute not only to our understanding of mast cell differentiation but also to Fc receptor ontogeny and stem cell development.

肥大细胞是直接疾病的主要效应细胞 超敏反应。 了解肥大细胞如何从原始细胞发育而来 血细胞可能对某些过敏症的治疗有影响 疾病。 因为肥大细胞可能与造血有独特的关系 干细胞，对肥大细胞分化早期事件的研究可能有 对一般造血作用具有更广泛的影响。 研究提出 在这项拨款申请中扩展了所开发的肥大细胞模型系统 在上一个补助期内。 该系统利用了以下事实： 承诺的祖先，而不是未承诺的祖先，可以对干做出反应 通过在甲基纤维素中产生肥大细胞集落来单独使用细胞因子。 人们持续关注 c-kit/stem 的不同角色 细胞因子受体-配体复合物可能在不同阶段发挥作用 分化可能对造血作用具有更广泛的影响 一般的。 本拨款申请中提出的研究扩展了 肥大细胞模型系统是在上一个资助期间开发的。 这 系统利用了承诺祖先的事实，但没有 未承诺的，可以通过产生单独的干细胞因子做出反应 甲基纤维素中的肥大细胞集落。 持续关注 c-kit/干细胞因子受体-配体的不同作用 复合体可能在分化的不同阶段发挥作用；然而， 本次拨款申请的主要重点是针对桅杆上的 Fc 受体 细胞及其祖细胞，因为最近的挑衅性发现 在肥大细胞中，FcRI 和 FcRIII 均与 γ 链二聚体相关 转导激活和细胞因子生物合成的信号。 我们是 对诱导自分泌 IL-3 的可能性特别感兴趣 循环可能是这些祖先的承诺状态的关键元素。 第一个具体目标是确定骨髓祖细胞是否 特别是那些可能属于早期肥大细胞谱系的细胞， FcRI 和 FcRIII，如果这些受体转导信号以诱导 自分泌 IL-3 环是承诺所必需的。 在第二个 特定目标、肥大细胞定向祖细胞或粒状肥大细胞 将测试 MLN 或腹膜冲洗以确定是否 FcRI/FcRIII 介导的信号传导影响自分泌的表达 细胞因子、GATA 转录因子或蛋白酶。 我们还将 确定是否有必要继续占据肥大细胞上的 FcRI 用于连续世代子细胞的持续分裂。 这 第三个具体目标利用了令人兴奋的新模型系统 造血作用的研究；胚胎干（ES）细胞培养。 一次 这些非常原始的细胞可以在培养物中分化， 将监测 FcRI/FcRIII 的信息和蛋白质的表达，以 与肥大细胞和自分泌 IL-3 的出现相关 诱导性。 这些实验可能揭示相似点或差异 本次赠款申请中提出的之间不仅应有助于 我们对肥大细胞分化的理解以及对 Fc 受体的理解 个体发育和干细胞发育。