GENETIC ANALYSIS OF CD18 AND ICAM-1 IN MICE AND HUMANS

小鼠和人类 CD18 和 ICAM-1 的遗传分析

基本信息

批准号：
2057143
负责人：
ERIC T SANDBERG
金额：
$ 9.18万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-07-01 至 1995-05-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2057143
关键词：
CD antigens cell adhesion molecules cellular immunity electroporation embryonic stem cell gene mutation genetic polymorphism human genetic material tag human tissue laboratory mouse leukocytes nucleic acid sequence polymerase chain reaction recombinant DNA southern blotting tissue /cell culture transplant rejection transplantation

项目摘要

Leukocyte and endothelial cell adhesion molecules are involved in critical pathways of the inflammatory response. Leukocyte adhesion deficiency is characterized by childhood presentation of indolent, recurrent infections of the skin, oropharynx, and respiratory tract. The molecular basis is defective production of CD18 leading to reduced cellular expression of leukocyte integrins. ICAM-1 is a ligand for one of the CD18 family of integrins. ICAM-1 is a member of the immunoglobulin supergene family and has been shown to play an important role in transplantation biology. In phase I of this proposal, I will join the efforts in Dr. Beaudet's laboratory aimed at obtaining murine models with mutations of CD18 and ICAM-1 to better delineate their biological functions. One germ-line mutation in CD18 has been obtained and others are being sought. I expect to be particularly involved in the introduction of mutations in the ICAM-1 gene using homologous recombination in embryonic stem (ES) cells. Mutant ES cells will be obtained using selection in tissue culture, screening with PCR, and confirmation by Southern blotting. Mutant cells will be used to establish germ-line transmission, and mutant mice will be characterized phenotypically. The effect of mutations in CD18 and ICAM-1 will be tested using transplantation with bone marrow and skin or cardiac tissue. In phase II of this project, I will focus on further biological studies of the mutant animals, and I will address the significance of potential genetic variation in these cell adhesion molecules in humans. The coding exons of CD18 and ICAM-1 will be sequenced in normal and high risk populations searching for amino acid polymorphisms of clinical significance. If found, the frequency of these amino acid polymorphisms in various clinical populations would be analyzed. Additional studies in the mice would focus on the effect of these mutations on common disease processes such as atherosclerosis, autoimmune disease, and inflammatory processes. A parallel goal of this project is preparation of the candidate for a career as an independent investigator. A series of graduate school courses focusing on cell and molecular biology will be taken for credit. An intense exposure to laboratory research is planned with 100% of time devoted to research training during the early years. The Department of Pediatrics has a long-term commitment to a faculty position in the Section of Allergy and Immunology.

白细胞和内皮细胞粘附分子参与关键炎症反应的途径。白细胞粘附不足是以儿童期表现为懒惰，反复感染的特征皮肤，口咽和呼吸道的。分子基础是 CD18的缺陷导致细胞表达降低白细胞整合素。 ICAM-1是CD18家族之一的配体整合素。 ICAM-1是免疫球蛋白超基因家族的成员，已被证明在移植生物学中起着重要作用。在该提案的第一阶段，我将加入Beaudet博士的努力实验室旨在获取具有CD18突变和的鼠模型 ICAM-1更好地描述其生物学功能。一个种菌已经获得了CD18中的突变，并且正在寻找其他突变。我希望特别参与ICAM-1中突变的引入使用胚胎干细胞中同源重组的基因。突变体 ES细胞将使用组织培养中的选择获得 PCR，并通过Southern印迹确认。突变细胞将用于建立种系传输，将表征突变小鼠表型。 CD18和ICAM-1中突变的效果将测试使用骨髓和皮肤或心脏组织的移植。在该项目的第二阶段，我将重点介绍突变动物，我将解决潜在遗传的重要性这些细胞粘附分子的变化。编码外显子 CD18和ICAM-1将在正常和高风险种群中进行测序寻找具有临床意义的氨基酸多态性。如果发现，这些氨基酸多态性在各种临床上的频率人口将进行分析。在小鼠中的其他研究将集中这些突变对常见疾病过程的影响，例如动脉粥样硬化，自身免疫性疾病和炎症过程。该项目的一个平行目标是为候选人做准备作为独立调查员的职业。一系列研究生院课程专注于细胞和分子生物学将获得信用。一个计划以100％的时间进行强烈接触实验室研究在早期致力于研究培训。部门儿科对本节的教师职位有长期的承诺过敏和免疫学。