Characterization of Lamprey B cells and Antibodies

项目摘要 七鳃鳗是现存的两种无颌脊椎动物之一，在研究鱼类的进化过程中占有重要地位。 适应性免疫系统建议的研究建立在以前的研究结果表明，互动的B-和 类T淋巴细胞是无颌和有颌动物适应性免疫系统的基本特征。 脊椎动物，尽管无颌脊椎动物产生可变淋巴细胞受体（VLR）用于抗原识别 通过将不完整的VLRA、VLRB和VLRC种系基因组合转化为完全组装的VLR， 使用邻近的富含亮氨酸的重复序列（LRR）作为模板的基因。先前的研究表明， 而VLRA和VLRC基因组装与胞苷脱氨酶1（CDA 1）在细胞中的表达一致， 胸腺等效鳃区，同时VLRB和CDA 2的表达主要发生在造血组织。 本文对VLRB B的发育、分布和功能进行了综合分析。 比如七鳃鳗体内的细胞及其抗体产物。第一个具体目标是阐明一代， VLRB抗体库的多样化和表达，特别强调新的 鉴定的CDA 2同种型。第二个具体目标是确定 七鳃鳗B样淋巴细胞的三种关键受体，即复合VLRB受体、IL-17 D受体和 BAFF/APRIL受体，通过使用互补和新的策略扩增可评估的VLRB 淋巴细胞群第三个具体目标是产生针对人肿瘤细胞的特异性VLRB抗体。 抗原并修饰它们用于诊断和潜在的治疗用途。这些研究的总体目标 是为了更好地了解无颌脊椎动物的B细胞生物学的基本方面，特别是 产生VLRB抗体多样性，并利用七鳃鳗抗体的优势用于生物医学 目的

PROJECT SUMMARY As one of two extant jawless vertebrates, lampreys occupy a pivotal position for study of the evolution of our adaptive immune system. The proposed studies build on previous findings indicating that interactive B- and T-like lymphocytes are fundamental features of the adaptive immune system in both jawless and jawed vertebrates, although jawless vertebrates generate variable lymphocyte receptors (VLR) for antigen recognition by combinatorial conversion of incomplete VLRA, VLRB and VLRC germ-line genes into fully assembled VLR genes using neighboring leucine-rich repeat (LRR) sequences as templates. Prior studies indicate that whereas VLRA & VLRC gene assemblies coincide with expression of cytidine deaminase 1 (CDA1) in the thymus equivalent gill region, concurrent VLRB and CDA2 expression occurs primarily in hematopoietic tissues. A comprehensive analysis is proposed to elucidate the development, distribution and function of the VLRB B- like cells and their antibody products in lampreys. The first specific aim is to elucidate the generation, diversification and expression of the VLRB antibody repertoire, with special emphasis on the role of newly identified CDA2 isoforms. The second specific aim is to define the composition and functional capabilities of three key receptors of lamprey B-like lymphocytes, namely the composite VLRB receptor, IL-17D receptor and BAFF/APRIL receptors, through the use of complementary and novel strategies to amplify the assessable VLRB lymphocyte population. The third specific aim is to generate specific VLRB antibodies against human tumor cell antigens and to modify them for diagnostic and potential therapeutic uses. The overall goals of these studies are to better understand basic aspects of B cell biology in a jawless vertebrate, in particular the mechanism of generating VLRB antibody diversity, and to exploit the advantages of lamprey antibodies for biomedical purposes.