GENETIC ASPECTS OF IMMUNODEFICIENCY

免疫缺陷的遗传方面

• 批准号: 3138492
• 负责人: MARY ELLEN CONLEY
• 金额: $ 14.68万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1993-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3138492
• 关键词: B lymphocyte; DNA methylation; T lymphocyte; Wiskott Aldrich syndrome; autoradiography; cell differentiation; cell sorting; cell transformation; complementary DNA; endonuclease; female; gel electrophoresis; gene mutation; genetic disorder diagnosis; genetic manipulation; genetic mapping; human tissue; hybridomas; hypogammaglobulinemia; immunodeficiency; immunogenetics; messenger RNA; molecular pathology; natural killer cells; nucleic acid probes; pancreatic ribonuclease; population genetics; severe combined immunodeficiency; sex linked trait

Carriers of the X-linked immunodeficiencies - severe combined immunodeficiency (SCID), Wiskott-Aldrich syndrome, hyper-IgM syndrome and X-linked lymphoproliferative syndrome-are normal by all immunologic parameters. We propose that the failure of these obligate heterozygotes to demonstrate any sign of these disorders is due to selective use of the X chromosome that does not carry the gene defect as the active X in all cell lines affected by the gene defect. If this hypothesis is correct, techniques that permit the distinction of the active and inactive X chromosomes will allow identification of the cell lines affected by the gene defects and provide a basis for carrier detection assays in these disorders. We have recently developed such a technique. Somatic cell hybrids are produced between purified T cells, B cells, NK cells or monocytes from a woman at risk and a chinese hamster cell line that is deficient in the X-linked enzyme HGPRT. Hybrids, which tend to lose human chromosomes, are grown in selective media so that only those hybrids that have retained the active human X chromosome will survive. DNA from these hybrids is analyzed, using an X-linked restriction fragment length polymorphism (RFLP) for which the woman is heterozygous. If the cell lineage used to make the hybrids is not affected by the gene defect, or if the woman studied is not a carrier of the disease in question, approximately half of the hybrids will use the maternally derived X chromosome as the active X and half will use the paternally derived one. However, if the woman studied is a carrier and the cell lineage studied is affected by the gene defect, then all the hybrids will use the X chromosome that does not carry the gene defect as the active X (non-random X chromosome inactivation). We intend to use this technique to determine the cell lineages affected by the gene defects listed above, and to provide carrier detection for these disorders. Our initial studies will focus on X-linked SCID for the following reasons: 1) determining the cell lines affected by this disorder will be particularly useful in clarifying the relationships between the various cell lines involved in the immune response; 2) there are no clinical or laboratory characteristics of X-linked SCID that distinguish it from other forms of SCID, making informed genetic counseling difficult; and 3) our ability to identify carriers of X- linked SCID will increase the number of informative individuals in affected pedigrees and thereby make it easier for us to map this disorder.

X连锁免疫缺陷的载体 - 严重的合并 免疫缺陷（SCID），Wiskott-Aldrich综合征，超基因 综合征和X连锁淋巴增生性综合征 - 正常 所有免疫学参数。 我们建议失败 这些强制性杂合子可以证明这些迹象 疾病是由于选择性使用X染色体的选择 在所有受影响的细胞系中，不带有基因缺陷为活性X 由于基因缺陷。 如果这个假设是正确的，那么技术是 允许主动和非活性X染色体的区别 将允许鉴定受基因影响的细胞系 缺陷并为这些载体检测分析提供了基础 疾病。 我们最近开发了这样的技术。 躯体 细胞杂种是在纯化的T细胞，B细胞，NK之间产生的 来自有危险的女人和中国仓鼠的细胞或单核细胞 缺乏X连锁酶HGPRT的细胞系。 倾向于失去人类染色体的杂种生长 选择性媒体，因此只有那些保留的杂种 活跃的人X染色体将生存。 来自这些的DNA 使用X连锁限制片段长度分析杂种 女性是杂合的多态性（RFLP）。 如果 用于制造杂种的细胞谱系不受 基因缺陷，或者如果被研究的女人不是 有关疾病，大约一半的杂种将使用 母体得出的X染色体，因为活性X和一半将 使用含父派生的一个。 但是，如果那个女人学习的是 载体和研究的细胞谱系受到基因的影响 缺陷，然后所有混合动力都将使用X染色体 不携带基因缺陷作为活性x（非随机x 染色体灭活）。 我们打算将此技术用于 确定列出的基因缺陷影响的细胞谱系 上面，并为这些疾病提供携带者检测。 我们的 初步研究将集中于以下的X连锁SCID 原因：1）确定受这种疾病影响的细胞系 在阐明 涉及免疫反应的各种细胞系； 2）那里 没有X连锁SCID的临床或实验室特征 将其与其他形式的SCID区分开，使知情的遗传 咨询困难； 3）我们识别X-载体的能力 链接的SCID将增加信息信息的信息 受影响的血统，从而使我们更容易映射 紊乱。