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MOLECULAR BASIS OF POLYCLONAL LYMPHOCYTE ACTIVATION

多克隆淋巴细胞激活的分子基础

基本信息

批准号：
3297506
负责人：
ROBERT DEANS
金额：
$ 10.53万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
1988
资助国家：
美国
起止时间：
1988-04-01 至 1991-03-31
项目状态：
已结题

项目摘要

This is a proposal to study the regulation of B lymphocyte activation at the molecular level, and particularly to gain insight into control mechanisms governing cell proliferation. A major focus of this study is the identification of the gene product defective in a lipopolysaccharide (LPS) non-responsive mouse line. This inbred of mice, C3H//HeJ, is genetically deficient in LPS- induced polyclonal spleen cell activation. This recessive mutation maps to a single locus (lps) on mouse chromosome 4. It is our long range goal to identify this gene and its product, and ultimately to restore competency for LPS responsiveness in mutant lymphocytes by gene re-introduction. Resting splenic lymphocytes respond to a variety of cell surface stimulation by undergoing general metabolic activation, maturation of immunoglobulin production from a surface receptor to a secreted molecule, and entry into the cell cycle. LPS is a potent polyclonal (antigen-independent) B cell activator, stimulating a high proportion (30%) of splenic B cells. B cell responses induced by LPS have common pathways of metabolic and cellular responses exhibited by antigen-induced B cell activation, and these events are currently poorly characterized at the molecular level. We have identified immunochemically a molecule (p-lps) which is unique to LPS-responsive B cells, and absent in LPS non- responsive B cells or spleen cells from C3H/HeJ mice. We have generated monoclonal antibodies (MoAb) which recognize this molecule, and use these antibodies to successfully screen a spleen cell gtII cDNA library. We propose to rigorously test these cDNA clones for their linkage to the lps allele by several distinct strategies. These anti-p-lps MoAb are valuable reagents to use in biochemically characterizing the p-lps molecule. We will determine its pattern of expression in various tissues and lymphocyte subpopulations. As well, we will study potential modification, both post-transcriptionally and post-LPS stimulation. We will investigate the association of p-lps with other molecules, with a particular focus on determining the involvement of p-lps in signal transduction pathways operating in B cell activation. The study of the lps gene and its product will yield insight into the molecular basis by which B cells are activated and lymphocyte proliferation responses induced in the immune response. This contributes to studies on abnormal B cell growth control in autoimmunity or neoplastic disease. This is a model system for an approach of gene complementation to restore inherited immune defects.

这是一项研究B淋巴细胞调节的提案在分子水平上激活，特别是为了获得洞察力进入细胞增殖的控制机制。一个专业本研究的重点是基因产物的鉴定脂多糖（LPS）无反应小鼠系中有缺陷。这种小鼠近交系 C3H//HeJ 在遗传上缺乏 LPS- 诱导多克隆脾细胞活化。这种隐性突变映射到小鼠 4 号染色体上的单个位点 (lps)。这是我们的长范围目标是识别该基因及其产物，并最终恢复突变体的 LPS 反应能力通过基因重新导入淋巴细胞。静息脾淋巴细胞对多种细胞表面做出反应通过一般代谢激活进行刺激，表面受体免疫球蛋白生产的成熟分泌分子，并进入细胞周期。脂多糖是一种有效的多克隆（不依赖于抗原）B 细胞激活剂，刺激高比例 (30%) 的脾 B 细胞。 B细胞 LPS 诱导的反应具有共同的代谢途径和抗原诱导的 B 细胞表现出的细胞反应激活，目前这些事件的特征还很不清楚分子水平。我们通过免疫化学方法鉴定出一种分子 (p-lps)，它是 LPS 反应性 B 细胞所特有，并且在 LPS 非反应性 B 细胞中不存在来自 C3H/HeJ 小鼠的反应性 B 细胞或脾细胞。我们有产生了识别这一点的单克隆抗体（MoAb）分子，并使用这些抗体成功筛选脾脏细胞gtII cDNA文库。我们建议严格测试这些 cDNA 克隆通过几个不同的方式与 lps 等位基因连接策略。这些抗 p-lps MoAb 是有价值的试剂，可用于 p-lps 分子的生化特征。我们将确定其在各种组织中的表达模式并淋巴细胞亚群。我们还将研究潜力转录后和 LPS 后的修饰刺激。我们将研究 p-lps 与其他分子，特别关注确定 p-lps 参与信号转导途径 B细胞激活。对 lps 基因及其产物的研究将深入了解 B细胞和淋巴细胞被激活的分子基础免疫反应中诱导的增殖反应。这有助于研究异常 B 细胞生长控制自身免疫或肿瘤性疾病。这是一个模型系统基因互补恢复遗传性免疫的方法缺陷。