MOLECULAR BASIS OF BLOOD GROUP ANTIGENS

目标是1)向血库社区介绍更多 一种准确、简便的检测红细胞抗原的方法 通过研究血型的分子基础鉴定抗体 重点是等位基因变异的识别。2)至 确定某些红细胞血型的结构/功能关系。 3)将特定的血型基因型与疾病过程联系起来，并 了解这种疾病的基础。为了实现这些目标，我们有 由五个项目组成的合作计划。四个项目 研究Rh的分子和细胞生物学(项目1)，Kell (项目2)、Duffy(项目3)和XG(项目4)血型和项目 5是一个临床组件，它将应用分子生物学程序来 改进血液中抗体和抗原的检测 银行参考实验室。项目1旨在构建一张物理地图 整个Rh基因座并决定其顺序、转录方向 以及主要Rh基因的表达。这将导致理解 许多Rh相关抗原的分子基础，这在血液中是重要的 输血、自身免疫性溶血性疾病和人的溶血性疾病 新生儿(HDN)。Kell血型系统不相容的情况也存在 输血困难和HDN。Kell抗原驻留在 表面暴露的糖蛋白与锌中性内肽酶同源。 项目2的一个目标是确定红细胞表面的可能作用 多肽酶。红细胞上的Duffy糖蛋白(GpFy)是一种趋化因子 受体，也是人类疟疾寄生虫的结合部位 间日疟原虫。达菲蛋白也在其他组织中发现，如 肾、肺、胸腺和脑。项目3旨在确定功能 通过对gpFy结构的研究，设计了 防止间日疟原虫入侵的方法。项目4将确定xG是否 血型参与了细胞间的相互作用。除了……之外 造血组织XG存在于成纤维细胞中，与一种 MIC2基因的产物。项目5将使用合成和重组 多肽和等位基因特异的转染体分析抗体 病人。Rh、Kell、Duffy和XG的分子基础信息 血型(项目1-4)也将用于引入新的程序 用于鉴定血型抗原。这将对Pre-Rate产生影响 出生时确定胎儿是否有患HDN的风险以及是否应该 适用于某些免疫的抗原阴性血液的筛查 患者，例如那些患有镰状细胞病的人。这五个项目是 由三个核心单位提供支持：行政、细胞培养和细胞 分类。细胞培养单位将为调查人员提供培训， 从造血组织中分离CD34+细胞，培养祖细胞 并准备用于原代造血培养的测试试剂。单元格 分选单元将通过流式细胞仪对造血干细胞进行分选和分析 细胞、承诺的祖细胞和其他具有 分化程度不同。总的来说，这5个项目提供了 研究血液分子和细胞生物学的综合方法 利用所学知识实现血库现代化 程序，并了解一些血型的功能及其 可能与疾病有关。

The objectives are 1) To introduce, to the blood banking community, more exact and convenient methods for determining red cell antigens and identifying antibodies by studying the molecular basis of blood groups with emphasis on the identification of allelic variations. 2) To determine structure/function relationships of some red cell blood groups. 3) To link specific blood group genotypes with disease processes and to understand the basis for the disease. To accomplish these goals we have a collaborative program composed of five projects. Four projects investigate the molecular and cell biology of Rh (Project 1), Kell (Project 2), Duffy (Project 3) and Xg (Project 4) blood groups and project 5 is a clinical component which will apply molecular biology procedures to improving the detection of antibodies and antigens as practiced in a blood bank reference laboratory. Project 1 aims to construct a physical map of the entire Rh locus and determine the order, transcriptional orientation and expression of the major Rh genes. This will lead to understanding the molecular basis of many Rh-related antigens, which is important in blood transfusion, autoimmune hemolytic disease and in hemolytic disease of the newborn (HDN). Kell blood group system incompatibilities also present difficulties in blood transfusion and HDN. Kell antigens reside on a surface exposed glycoprotein with homology to zinc neutral endopeptidases. An aim of Project 2 is to determine the possible role of red cell surface peptidases. Duffy glycoprotein (gpFy), on red cells, acts as a chemokine receptor and also as a binding site for the human malarial parasite Plasmodium vivax. Duffy protein is also found in other tissues such as kidney, lung, thymus and brain. Project 3 aims to determine the function of gpFy in these tissues and, by studying the structure of gpFy, design ways to prevent P. vivax invasion. Project 4 will determine whether Xg blood group is involved in cell interactions. In addition to hematopoietic tissues Xg is found in fibroblasts and is homologous to a product of the MIC 2 gene. Project 5 will use synthetic and recombinant peptides, and allelic-specific transfectants to analyze antibodies in patients. Information on the molecular basis of Rh, Kell, Duffy and Xg blood groups (Projects 1-4) will also be used to introduce new procedures for the identification of blood group antigens. This will impact on pre- natal determination of whether a fetus is at risk for HDN and should be useful in screening for antigen-negative blood for certain immunized patients, such as those with Sickle Cell disease. The five projects are supported by three Core units; administrative, cell culture and cell sorting. The cell culture unit will provide training to investigators, isolate CD34+ cells from hematopoietic tissues, culture progenitor cells and prepare tested reagents for primary hematopoietic cultures. The cell sorting unit will sort and analyze, by flow cytometry, hematopoietic stem cells, committed progenitor cells and other hematopoietic cells with different degrees of differentiation. Overall these 5 projects provide an integrated approach to studying the molecular and cell biology of blood groups and applying the knowledge gained to modernize blood banking procedures and understand the functions of some blood groups and their possible relations to diseases.