ISOLATION OF THE CHEDIAK HIGASHI IMMUNE DEFICIENCY GENE

CHEDIAK HiGASHI 免疫缺陷基因的分离

• 批准号: 2667769
• 负责人: Frederick s Southwick
• 金额: $ 20.13万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2667769
• 关键词: Chediak Higashi syndrome; alleles; artificial chromosomes; chromosome aberrations; clinical research; gene complementation; gene expression; gene mutation; gene rearrangement; genetic mapping; human genetic material tag; human subject; immunodeficiency; immunogenetics; laboratory mouse; molecular cloning; northern blottings; nucleic acid hybridization; nucleic acid sequence; polymerase chain reaction; pulsed field gel electrophoresis; sequence tagged sites; southern blotting

Chediak-Higashi syndrome is a primary immune deficiency disease of human (CHS) and mouse (beige, bg). Affected individuals have defective granulocyte and Natural Killer cell activity, pathognomonic giant vesicles in many cell types, and typically die in childhood of infection or malignancy. There is strong evidence that human CHS is homologous to the bg locus in the mouse. Since attempts to identify the genetic defect in CHS directly have been thwarted by absence of chromosomal assignment of CHS or pedigrees adequate for linkage analysis, we have undertaken positional cloning of bg as an antecedent to identification of the CHS gene. Positional cloning is an approach to identifying a disease gene based solely on its chromosomal location, without regard to etiology. Our preliminary experiments have: 1. Localized bg to a 0.24+/-0.17 centiMorgan interval on mouse chromosome 13; Identified a genomic rearrangement within this interval in the SB/LeJ-bg mouse; 3. Characterized 3 yeast artificial chromosomes (YACs) which may span this rearrangement. We propose to identify the defective gene in the bg mouse by examining expressed sequences derived from these YACs for mutations. The human homolog of the bg gene will be cloned and sequenced in CHS patients and normal individuals to identify CHS mutations. Isolation of the CHS gene will permit prenatal genetic diagnosis, enabling institution of treatment for CHS in infancy. Identification of the bg gene will facilitate understanding of mechanisms of regulation of protein trafficking to lysosomes, and of the contribution of vesicular sorting to immune functions such as cellular cytotoxicity. Given the selective impairment of certain immune responses evident in bg mice, identification of the molecular basis of this disease may suggest a novel approach to immune modulation, and imply therapies for CHS which are not currently being considered, and will provide a starting point for gene therapy of CHS.

Chediak-Higashi综合征是人类的主要免疫缺陷疾病 （CHS）和鼠标（米色，bg）。受影响的人有缺陷 粒细胞和天然杀伤细胞活性，病理巨囊泡 在许多细胞类型中，通常死于感染的儿童或 恶性。有充分的证据表明，人类CHS与 鼠标中的BG基因座。由于试图识别遗传缺陷 通过缺乏染色体分配，CHS直接受到挫败 CHS或谱系足以进行连锁分析，我们已经承担了 BG的位置克隆作为CHS识别的先例 基因。 位置克隆是鉴定疾病基因的一种方法 仅基于其染色体位置，而无需病因。我们的 初步实验具有：1。局部BG至0.24 +/- 0.17厘米 小鼠染色体13的间隔；确定了基因组重排 此间隔在SB/LEJ-BG鼠标中； 3。表征3个酵母人工 可能跨越此重排的染色体（YAC）。 我们建议 通过检查表达的BG小鼠中的缺陷基因 从这些YAC衍生出来突变的序列。人类的同源 BG基因将在CHS患者和正常的CHS患者中进行克隆和测序 个人识别CHS突变。 CHS基因的隔离将 允许产前遗传诊断，使治疗机构 婴儿期。 BG基因的识别将有助于 了解蛋白质运输调节机制 溶酶体，以及囊泡分选对免疫的贡献 诸如细胞细胞毒性之类的功能。考虑到选择性损害 在BG小鼠中明显的某些免疫反应，鉴定 该疾病的分子基础可能表明一种新颖的免疫方法 调节，暗示当前尚未正在的CHS的疗法 考虑到CHS基因治疗的起点。