Comparison of B Cell Differentiation in Mice and Humans

小鼠和人类 B 细胞分化的比较

• 批准号: 7918590
• 负责人: Max Dale Cooper
• 金额: $ 25.06万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-12 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7918590
• 关键词: B cell differentiation; B cell repertoire; B-Cell Development; B-Lymphocytes; CD19 gene; CD32 Antigens; Cell Line; Cell Lineage; Cell Surface Receptors; Cell model; Cells; Clinical; Differentiation and Growth; Dominant-Negative Mutation; Event; Gene Expression; Gene Expression Profile; Generations; Genes; Genetic Programming; Human; IL7R gene; Immunofluorescence Immunologic; Immunoglobulin D; Immunoglobulin M; Interleukin-7; Ligation; Light; Lymphoid; Lymphopoiesis; Methods; Modeling; Modification; Molecular Profiling; Mus; Participant; Pathway interactions; Process; Receptors, Antigen, B-Cell; Stromal Cell-Derived Factor 1; Stromal Cells; TNFRSF5 gene; Testing; Transgenes; cell growth; comparative; novel; progenitor; programs; receptor; research study; surrogate light chain

This proposal focuses on unresolved issues of early B lineage differentiation in mice and, especially, in humans. (Aim 1) A novel model of human B cell development has been identified, the EU12 cell line, in which CD34+/|a heavy chain" pro-B cells spontaneously undergo step-wise differentiation to become IgM+/IgD+ B cells. This clone will be used to (i) test the hypothesis that intraclonal V(D)Jrecombinatorial diversification contributes to the efficient generation of a primary B cell repertoire, (ii) define the changes in gene expression profile during B lineage differentiation, (iii) test the effects of modifying newly-identified and previously-recognized B lineage genes in pro-B cells by sense, antisense and dominant-negative transgenes, and (iv) examine the effects of ligating cell surface receptors (preBCR, BCR, IL7R, CD19, CD32, and CD40) on growth and differentiation of these B lineage cells. (Aim 2) A recently-developed, highly- amplified immunofluorescence method will be used to identify pro-B, pre-B and B cell receptor components on primary B lineage cells to define similarities and differences in the human and mouse B cell differentiation programs. After redefinition of when and where the \i heavy chains, surrogate light chains, icA,light chains, and Igo/p are expressed during B lineage differentiation in both species, pro-B, pre-B, and B cell subpopulations will be isolated for comparative analysis of their gene expression profiles and differentiation potential. The latter experiments will incorporate new information obtained in the analysis of differentiation- related changes in the gene expression profile of EU12 cells. (Aim 3) Complementary ex vivo models will be used to elaborate early B lineage differentiation events in mice and humans. An IL-7 producing stomal cell model will be employed to delineate early events in mouse B lineage differentiation. Mouse stromal cells, with and without modification by human stromal cell-derived factor 1 (SDF-1), IL-7 and Fey receptor transgenes, will be employed to evaluate the capacity of human lymphoid progenitors to undergo B lineage differentiation. This comparative analysis will define novel features of B cell differentiationthat are likely to have significant clinical implications.

该提案重点关注小鼠早期 B 谱系分化的未解决问题，特别是在 人类。 （目标 1）已确定人类 B 细胞发育的新模型，即 EU12 细胞系， 其中 CD34+/|a 重链”pro-B 细胞自发地经历逐步分化成为 IgM+/IgD+ B 细胞。该克隆将用于 (i) 检验克隆内 V(D)J 重组的假设 多样化有助于原代 B 细胞库的有效生成，(ii) 定义 B 谱系分化过程中的基因表达谱，(iii) 测试修饰新鉴定的效果 以及先前通过有义、反义和显性失活在原 B 细胞中识别的 B 谱系基因 (iv) 检查连接细胞表面受体（preBCR、BCR、IL7R、CD19、CD32、 和 CD40）对这些 B 谱系细胞的生长和分化的影响。 （目标 2）最近开发的、高度 放大免疫荧光法将用于鉴定pro-B、pre-B和B细胞受体成分 对原代 B 谱系细胞进行研究，以确定人类和小鼠 B 细胞分化的相似性和差异 程序。重新定义\i重链、替代轻链、icA、轻链的时间和地点后， 和 Igo/p 在两个物种（pro-B、pre-B 和 B 细胞）的 B 谱系分化过程中表达 将分离亚群以对其基因表达谱和分化进行比较分析 潜在的。后面的实验将结合在分化分析中获得的新信息- EU12细胞基因表达谱的相关变化。 （目标 3）互补的离体模型将是 用于阐述小鼠和人类的早期 B 谱系分化事件。产生 IL-7 的造口细胞 模型将用于描述小鼠 B 谱系分化的早期事件。小鼠基质细胞， 经或未经人基质细胞衍生因子 1 (SDF-1)、IL-7 和 Fey 受体修饰 转基因，将用于评估人类淋巴祖细胞进行 B 谱系的能力 差异化。这种比较分析将定义 B 细胞分化的新特征，这些特征可能 具有重要的临床意义。