MUTANTS FOR CELL ADHESION MOLECULES

细胞粘附分子的突变体

• 批准号: 2067082
• 负责人: ARTHUR L. BEAUDET
• 金额: $ 22.87万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 1996-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2067082
• 关键词: atherosclerosis; cell adhesion; cell adhesion molecules; embryo /fetus tissue /cell culture; experimental allergic encephalomyelitis; gene mutation; genetic manipulation; genetic mapping; germ cells; heterozygote; histocompatibility; homozygote; inflammation; integrins; laboratory mouse; monoclonal antibody; mutant; nucleic acid sequence; polymerase chain reaction; protein structure function; restriction mapping; southern blotting; surface antigens; systemic lupus erythematosus; tissue /cell culture; transfection; transplantation immunology; transposon /insertion element

Numerous cell adhesion molecules found on leukocytes, endothelial cells, and platelets are involved in a central way in many biological functions and disease processes. These cell adhesion molecules include members of the immunoglobulin gene superfamily, integrins, selectins, and other proteins. This project focuses particularly on the beta-2 subunit of leukocyte integrins (CD18), intercellular adhesion molecule-1 (ICAM-1), and granule membrane protein-140 (GMP-140). The overall goals of this project are to generate mutations in the CD18, ICAM-1, and GMP-140 genes in the mouse using homologous recombination in embryonic stem (ES) cells. These mouse mutants would be used to study the role of these proteins in normal biological function and in disease. There is substantial evidence that these proteins play central roles in processes such as inflammation, autoimmune disease, infectious susceptibility, transplantation rejection, and perhaps atherosclerosis. The murine genes would be cloned and characterized in sufficient detail to prepare constructs for homologous recombination in ES cells. The CD18, ICAM-1, and GMP-140 genes would be interrupted by homologous recombination in ES cells using insertion and/or replacement vectors, tissue culture selection, screening with the polymerase chain reaction (PCR), and confirmation of homologous recombination with Southern blotting. Mutant ES cells would be introduced into mouse blastocysts to obtain chimeras and subsequently bred into the mouse germ-line. The phenotype of heterozygous and homozygous mutant mice will be examined clinically, pathologically, and physiologically. The effect of mutant phenotypes on murine autoimmune processes will be studied using model systems such as experimental allergic encephalomyelitis, lupus erythematosus, and diabetes mellitus. The effects of the mutant phenotypes on murine atherosclerosis will be examined using mice with putative atherosclerosis genotypes. Mutant mice will be made available on a collaborative basis and rapidly on an unlimited basis to other laboratories in order to facilitate analysis of other biological features of any mutant animals. There will be a sharp focus on obtaining null mutations in CD18, ICAM-1, and GMP-140 and on introducing these into the mouse germ-line in the first 1-3 years of the project.

在白细胞、内皮细胞、 而血小板在许多生物功能中起着中心作用 和疾病过程。这些细胞黏附分子包括 免疫球蛋白基因超家族、整合素、选择素等 蛋白质。这个项目特别关注的是β-2亚基。 白细胞整合素(CD18)、细胞间黏附分子-1(ICAM-1)和 颗粒膜蛋白140(GMP-140)。这个项目的总体目标是 是在CD18、ICAM-1和GMP-140基因中产生突变 在胚胎干细胞中使用同源重组的小鼠。这些 小鼠突变体将被用来研究这些蛋白在正常 生物学功能和在疾病中。有确凿的证据表明 这些蛋白质在炎症等过程中发挥核心作用， 自身免疫性疾病，感染易感性，移植排斥反应， 或许还有动脉粥样硬化。小鼠的基因将被克隆并 具有足够的细节特征，以准备用于同源的 在ES细胞中重组。CD18、ICAM-1和GMP-140基因将是 通过插入和/或在ES细胞中的同源重组而中断 替换载体，组织培养选择，用 聚合酶链式反应(PCR)和同源基因的确认 与Southern blotting重组。突变的ES细胞将被引入 转化为小鼠囊胚以获得嵌合体，然后培育成 小鼠的生殖系。杂合子和纯合子突变小鼠的表型 将进行临床、病理和生理检查。这个 突变表型对小鼠自身免疫过程的影响将被研究 使用模型系统，如实验性变态反应性脑脊髓炎、狼疮 红斑狼疮和糖尿病。突变表型的影响 关于小鼠动脉粥样硬化的研究将使用假定的 动脉粥样硬化基因分型。突变小鼠将在 在协作的基础上，在不受限制的基础上迅速与其他实验室 为了便于分析任何突变体的其他生物学特征 动物。将把重点放在获得CD18零突变上， ICAM-1和GMP-140，并将它们导入小鼠胚系 项目的头1-3年。