IG GENETICS--ONTOGENY AND DIFFERENTIATION OF CELLS OF THE RABBIT IMMUNE SYSTEM

IG遗传学--兔免疫系统细胞的个体发育和分化

• 批准号: 3790652
• 负责人: R G MAGE
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3790652
• 关键词: B lymphocyte; T cell receptor; bone marrow; cell differentiation; gene expression; gene rearrangement; genetic strain; glycoproteins; gut associated lymphoid tissue; immune complex; immune system; immunogenetics; immunoglobulin D; immunoglobulin M; immunoglobulin genes; laboratory rabbit; lectin; lymphocyte; mutant; nucleic acid repetitive sequence; oligosaccharides; recombinase; spleen

We use techniques of classical immunogenetics and of molecular biology to study the genetics of rabbit immunoglobulins (Igs ), T cell receptors (Tcr), and related genes such as the recombinase activating genes RAG-1 and RAG-2 which are necessary for gene rearrangements to occur during lymphocyte development. We investigate the development of germinal centers and the regulated expression and sequence diversification of Ig genes during lymphoid cell development. Four VH-CH recombination sites map 3' of the VH genes and 5' of the JH genes. There appear to be two or more "hot-spots of recombination" within or near stretches of repetitive DNA in the DH-containing region of the rabbit. A fifth recombinant (R7K) has a site that maps 3' of the exons encoding the membrane terminus of the IgM heavy chain (Cmu) and 5' of Cgamma. The exact site of this recombination and its possible localization in a region homologous to the region in other species containing exons for the IgD heavy chain is currently under investigation. In contrast to most mammals studied, membrane-IgD has not been definitively identified on rabbit B cells. We have identified two surface immunoglobulin complexes on rabbit peripheral blood B cells that possess heavy chains of similar apparent molecular size. These may represent complexes with rabbit IgM and IgD. Four proteins are found noncovalently associated with the mIg in each of the receptors. These include 42, 37 and 36 kD proteins that may be the rabbit homologues of murine Ig-beta (B29) and Ig-gamma and Ig-alpha (mb-1). These proteins are found associated as 75 kd and 115 kD heteromeric complexes. Two larger glycoproteins (100 and 150 kD) may be specifically associated with the non-mu receptor. The Ig-associated proteins vary in B-lymphocytes isolated from spleen, blood, bone marrow and gut associated lymphoid tissue (GALT). Lectins are allowing us to dissect rabbit GALT into subcompartments based on local accumulations of cells expressing particular oligosaccharides. Normal and VH-expression mutant ali rabbits show similar staining patterns;the defect in ali rabbits which leads to the expression of a different set of VH genes does not appear to affect the general architecture of GALT.

我们使用经典免疫遗传学和分子生物学技术 研究兔免疫球蛋白 (Igs)、T 细胞受体的遗传学 (Tcr) 和相关基因，例如重组酶激活基因 RAG-1 和 RAG-2，它们是基因重排发生所必需的 淋巴细胞发育。我们研究生发中心的发展 以及 Ig 基因的表达调控和序列多样化 在淋巴细胞发育过程中。 四个 VH-CH 重组位点图 3' VH基因的5'和JH基因的5'。似乎有两个或更多 重复 DNA 片段内或附近的“重组热点” 兔子的含有 DH 的区域。第五种重组体（R7K）具有 映射编码 IgM 膜末端的外显子 3' 的位点 重链 (Cmu) 和 Cgamma 的 5'。 这种重组的确切位点 及其可能定位于与该区域同源的区域 其他含有 IgD 重链外显子的物种目前正在研究中 调查。 与大多数研究的哺乳动物相比，膜 IgD 并没有 已在兔 B 细胞上得到明确鉴定。 我们已经确定了两个 兔外周血 B 细胞表面免疫球蛋白复合物 具有相似表观分子大小的重链。 这些可能 代表与兔 IgM 和 IgD 的复合物。 发现四种蛋白质 与每个受体中的 mIg 非共价结合。这些 包括 42、37 和 36 kD 蛋白质，可能是兔同源物 鼠 Ig-beta (B29) 以及 Ig-gamma 和 Ig-alpha (mb-1)。这些蛋白质是 发现关联为 75 kD 和 115 kD 异聚复合物。 两个较大的 糖蛋白（100 和 150 kD）可能与 非mu受体。 B 淋巴细胞中的 Ig 相关蛋白有所不同 从脾脏、血液、骨髓和肠道相关淋巴液中分离 组织（GALT）。 凝集素使我们能够将兔 GALT 解剖成 基于表达细胞的局部积累的子区室 特别是低聚糖。 正常和 VH 表达突变 ali 兔 显示相似的染色模式；阿里兔中的缺陷导致 一组不同的 VH 基因的表达似乎不会影响 GALT 的总体架构。