MUTANTS FOR CELL ADHESION MOLECULES

细胞粘附分子的突变体

• 批准号: 3147222
• 负责人: ARTHUR L. BEAUDET
• 金额: $ 20.96万
• 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/3147222
• 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， 血小板在许多生物功能中起着重要作用 和疾病过程。 这些细胞粘附分子包括 免疫球蛋白基因超家族、整合素、选择素等 proteins. 该项目特别关注β-2亚基， 白细胞整合素（CD 18）、细胞间粘附分子-1（ICAM-1）和 颗粒膜蛋白140（GMP-140）。 本项目的总体目标 是在CD 18，ICAM-1和GMP-140基因中产生突变， 小鼠胚胎干细胞（ES细胞）中的同源重组。 这些 小鼠突变体将用于研究这些蛋白质在正常细胞中的作用。 生物学功能和疾病。 有大量证据表明 这些蛋白质在诸如炎症的过程中起中心作用， 自身免疫性疾病，感染易感性，移植排斥， 也可能是动脉粥样硬化。 小鼠基因将被克隆， 足够详细地表征以制备用于同源的 ES细胞中的重组。 CD 18、ICAM-1和GMP-140基因可能是 通过使用插入和/或插入在ES细胞中的同源重组中断， 替换载体，组织培养选择，筛选 聚合酶链反应（PCR），并确认同源 Southern blotting检测。 突变的胚胎干细胞将被引入 植入小鼠胚泡中以获得嵌合体，并随后繁殖到 小鼠生殖系。 杂合和纯合突变小鼠的表型 将进行临床、病理和生理检查。 的 将研究突变表型对小鼠自身免疫过程的影响 使用模型系统如实验性过敏性脑脊髓炎、狼疮 红斑和糖尿病。 突变表型的影响 对小鼠动脉粥样硬化的影响将使用具有假定的 动脉粥样硬化基因型 突变小鼠将在一个 合作的基础上，并迅速在不受限制的基础上，以其他实验室 为了便于分析任何突变体的其他生物学特征， 动物 将有一个尖锐的重点是获得无效突变的CD 18， ICAM-1和GMP-140，并将其引入小鼠生殖系， 项目的前1-3年。