MOLECULAR BASIS OF POLYCLONAL LYMPHOCYTE ACTIVATION

多克隆淋巴细胞激活的分子基础

• 批准号: 3297504
• 负责人: ROBERT DEANS
• 金额: $ 11.16万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 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
• 来源: https://reporter.nih.gov/project-details/3297504
• 关键词: B lymphocyte; alleles; chromosome translocation; fluorescent dye /probe; gene expression; genetic library; hybrid cells; immune tolerance /unresponsiveness; immunochemistry; laboratory mouse; laboratory rabbit; leukocyte activation /transformation; linkage mapping; lipopolysaccharides; membrane proteins; molecular cloning; monoclonal antibody; mutant; nucleic acid sequence; protein structure; tissue /cell culture

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.

这是研究B淋巴细胞调节的一个建议 在分子水平上激活，特别是为了了解 转化为控制细胞增殖的机制。 一个主要 本研究的重点是基因产物的鉴定 在脂多糖（LPS）无应答小鼠系中有缺陷。 这种近交系小鼠，C3 H//HeJ，在LPS- 诱导多克隆脾细胞活化。 这种隐性突变 定位于小鼠4号染色体上的单个基因座（LPS）。 这是我们的长期 我们的目标是识别这个基因及其产物，并最终 恢复突变体的LPS应答能力 通过基因再引入的淋巴细胞。 静息脾淋巴细胞对多种细胞表面 通过经历一般代谢活化的刺激， 从表面受体产生免疫球蛋白的成熟 转化为分泌的分子，并进入细胞周期。 LPS是一种 有效的多克隆（抗原非依赖性）B细胞活化剂， 刺激高比例（30%）的脾B细胞。 B细胞 LPS诱导的反应有共同的代谢途径， 和抗原诱导的B细胞表现出的细胞应答 激活，这些事件目前的特征很差， 分子水平。 我们已经通过免疫化学方法鉴定了一种分子（p-LPS）， 对LPS敏感的B细胞是唯一的，而在LPS非敏感的B细胞中不存在。 来自C3 H/HeJ小鼠的应答性B细胞或脾细胞。 我们有 产生的单克隆抗体（MoAb）， 分子，并使用这些抗体成功地筛选脾脏 细胞gtII cDNA文库。 我们建议严格测试这些cDNA 通过几种不同的方法将它们与LPS等位基因连锁 战略布局 这些抗β-脂蛋白单克隆抗体是有价值的试剂， 生物化学表征p-LPS分子。 我们将 确定其在各种组织中的表达模式， 淋巴细胞亚群 同时，我们将研究潜在的 转录后和LPS后的修饰 刺激. 我们将研究p-lps与 其他分子，特别侧重于确定 p-LPS参与细胞内信号转导通路， B细胞活化。 对lps基因及其产物的研究将使我们对lps的发病机制有更深入的了解。 B细胞被激活的分子基础和淋巴细胞 免疫应答中诱导的增殖应答。 这 有助于研究异常B细胞生长控制， 自身免疫或肿瘤疾病。 这是一个用于 基因互补恢复遗传免疫的途径 缺陷