MOLECULAR PHENOTYPING BY CYTOMETRY--CYTOKINE EXPRESSION

描述：拟议研究的目的是开发方法 同时进行免疫表型和分子表型的细胞 评估细胞因子受体和配体的表达。造血细胞 来源于一小群祖细胞，并分化为 多潜能干细胞转化为致力于扩增的祖细胞 成为有功能的末端细胞。已经产生了对某些病毒的抗体 造血细胞膜上的蛋白质，但不是全部。 因为每个子集都显示了唯一的蛋白质谱库 分化途径，研究人员试图能够分离出 并用流式细胞仪鉴定特定的细胞群。这个 研究人员假设造血细胞亚群表达 与配体结合的独特细胞因子受体谱系 调节这些细胞的增殖和分化。至 了解此差异化过程的功能方面 建议开发免疫表型和免疫表型所需的程序 分子表型。分子表型是一个用来描述 确定细胞内特定核酸序列的过程 手机。这两种方法都是必要的，因为特定的抗体 以确定可用的某些细胞类型，但可用的细胞很少 用于测量细胞因子配体和仅一些配体受体。分子 原位聚合酶链式反应与免疫表型相结合的表型鉴定尚不成熟 完全归结于实践，因此，一些开发工作也 这是必要的。核酸序列的检测将通过以下方式进行 悬浮液和原位荧光原位杂交 聚合酶链式反应。该提案强调使用流式细胞术进行检测， 然而，显微成像也将被使用。这些实验是 旨在提供有助于理解正常情况的信息 造血术。一种可靠的分子检测方法的发展 用流式细胞术进行表型鉴定将对基础、 翻译和临床研究。 这项研究有三个目标。一是制定可靠的 一种可组合的流式细胞仪分子表型分析方法 进行免疫表型鉴定。第二是应用这项技术来定义 造血细胞亚群表达的特异性细胞因子受体 祖细胞。第三个目标是定义由 T细胞和单核细胞的亚群。方法和结果将开启一个新的 细胞功能的询问方法。

DESCRIPTION: The purpose of the proposed research is to develop methods to simultaneously immunophenotype and molecular phenotype cells to evaluate cytokine receptor and ligand expression. Hematopoietic cells are derived from a small pool of progenitor cells and differentiate from multipotential stem cells into committed progenitor cells that expand to become functional end cells. Antibodies have been produced to some of the proteins on the membrane of hematopoietic cells but not all of them. Since a unique repertoire of proteins is displayed for each subset along the differentiation pathway, investigators seek to be able to separate and identify specific cell populations using flow cytometry. The investigators hypothesize that hematopoietic cell subsets express the unique repertoire of cytokine receptors whose engagement by ligands regulates both proliferation and differentiation of these cells. To understand the functional aspects of this differentiation process it is proposed to develop the procedures necessary to both immunophenotype and molecular phenotype. Molecular phenotyping is a term applied to describe the process of determining specific nucleic acids sequences inside a cell. Both of these methods are necessary because the specific antibodies to identify certain cell types that are available, yet few are available for measuring cytokine ligands and only some ligand receptors. Molecular phenotyping by in situ PCR combined with immunophenotyping is not yet completely reduced to practice, therefore, some development work is also necessary. The detection of nucleic acid sequences will be performed by fluorescence in situ hybridization in suspension as well as by in situ PCR. The proposal emphasizes the use of flow cytometry for detection, however, microscopic imaging will be used as well. The experiments are designed to provide information that will help understand normal hematopoiesis. The development of a reliable method for molecular phenotyping by flow cytometry will have an enormous impact on basic, translational and clinical research. There are three goals to this study. The first is to develop a reliable method for molecular phenotyping by flow cytometry that can be combined with immunophenotyping. The second is to apply this technology to define the specific cytokine receptor expressed by subsets of hematopoietic progenitor cells. The third goal is to define the ligands produced by subsets of T-cells and monocytes. The methods and results will open a new approach to the interrogation of cellular function.