Human B cell homeostasis in a new transgenic xenochimera

新型转基因异种嵌合体中的人类 B 细胞稳态

• 批准号: 6705698
• 负责人: Robert Thomas Woodland
• 金额: $ 25.17万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6705698
• 关键词: B lymphocyte; SDS polyacrylamide gel electrophoresis; antibody; biological models; cell differentiation; cell proliferation; enzyme linked immunosorbent assay; gene expression; genetically modified animals; hematopoietic stem cells; homeostasis; immunoglobulins; laboratory mouse; model design /development; polymerase chain reaction; western blottings; xenotransplantation

DESCRIPTION (provided by applicant): Xenochimeric animals produced by infusing human hematopoietic cells into immunodeficient mice have provided a valuable surrogate small animal model to study human lymphocyte development, maintenance, and function in vivo. Attempts to effect complete human B cell reconstitution, however, in non-obese diabetic (NOD) mice with scid or rag mutations have met with varying success. Functional long-lived B cells fail to develop when immature cord blood cells are used to reconstitute NOD/scids. Likewise, B cells engraft poorly if at all when mature human peripheral blood lymphocytes are transferred. Peripheral B cell maturation and survival are completely dependent on the newly described B cell stimulator (BLyS), also known as BAFF, TALL and THANK. Because peripheral B cell maturation ceased in the chimeras at a BlyS dependent checkpoint, we hypothesized murine BLyS in the NOD recipients is limiting, because of quantity, location or species restrictions for utilization. We present preliminary experiments to show robust B cell engraftment in NOD-rag-/-Pfp-/- recipients of human PBL supplemented daily with recombinant human BLyS. No human B cells develop in mice without BLyS supplement. From these data, we formulated the hypothesis that the failure of human B cells to develop and survive in xenochimeras is due to BlyS deficiency. We propose the following experimental aims to develop a stable transfer environment for human B cells, and to examine in vivo the function of BLyS on human peripheral B cell homeostasis. Aim 1. To develop a NOD-rag-/-Pfp-/- transgenic mouse that can produce recombinant human BlyS in a regulated fashion. Aim 2. To determine if human BLyS from a transgene will support the peripheral maturation of long lived naive B cells from human cord blood cells; and to determine if human BLyS will support the survival of mature peripheral B cells. Aim 3. To determine if the B cells in the NOD-rag-/-Pfp-/- BLyS transgenics can be induced to mount recall or de novo antibody responses to challenge with vaccine. The new transgenic mice we propose to produce will provide both new insights and a new small animal model system for determining human B cell development and function in vivo. Furthermore, These transgenic mice will form the basis of a new test system to study the efficacy of adjuvants and vaccines currently being developed against agents of bioterrorism.

描述（由申请人提供）：通过将人造血细胞输注到免疫缺陷小鼠中产生的异种嵌合动物提供了一种有价值的替代小动物模型，用于研究体内人淋巴细胞发育、维持和功能。然而，在具有scid或rag突变的非肥胖糖尿病（NOD）小鼠中实现完整的人B细胞重建的尝试已经取得了不同的成功。当未成熟的脐带血细胞用于重建NOD/SCIDS时，功能性长寿的B细胞不能发育。同样地，当转移成熟的人外周血淋巴细胞时，B细胞即使植入也很差。外周B细胞的成熟和存活完全依赖于新描述的B细胞刺激因子（BLyS），也称为BAFF、TALL和THANK。由于嵌合体中外周B细胞成熟在BlyS依赖性检查点停止，我们假设NOD受体中的鼠BLyS是有限的，因为数量、位置或物种限制。我们提出了初步实验，以显示每日补充重组人BLyS的人PBL的NOD-rag-/-Pfp-/-受体中稳健的B细胞植入。在没有BLyS补充的小鼠中没有人B细胞发育。根据这些数据，我们提出了一个假设，即人类B细胞在异种嵌合体中发育和存活的失败是由于BlyS缺陷。我们提出了以下实验目的，以开发一个稳定的转移环境，为人类B细胞，并在体内检查的功能BLyS对人类外周B细胞的稳态。 目标1.目的：建立一种能稳定表达重组人BlyS的NOD-rag-/-Pfp-/-转基因小鼠。 目标2.确定来自转基因的人BLyS是否将支持来自人脐带血细胞的长寿幼稚B细胞的外周成熟;并确定人BLyS是否将支持成熟外周B细胞的存活。 目标3.确定NOD-rag-/-Pfp-/- BLyS转基因中的B细胞是否可被诱导产生对疫苗攻击的回忆或从头抗体应答。 我们提出的新的转基因小鼠生产将提供新的见解和一个新的小动物模型系统，以确定人类B细胞的发展和功能在体内。此外，这些转基因小鼠将构成一个新的测试系统的基础，以研究目前正在开发的针对生物恐怖主义制剂的佐剂和疫苗的功效。