DEVELOPMENT OF HUMAN IMMUNOGLOBULIN V GENE REPERTOIRES

人类免疫球蛋白 V 基因库的开发

• 批准号: 2100048
• 负责人: DAVID F FRIEDMAN
• 金额: $ 8.2万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-01-20 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2100048
• 关键词: B lymphocyte; antibody formation; child (0-11); flow cytometry; gene induction /repression; gene rearrangement; genetic library; human genetic material tag; human subject; immunoglobulin genes; leukocyte activation /transformation; nucleic acid sequence; oligonucleotides

My long term goal is to pursue a career in Academic Pediatrics which combines laboratory research and clinical activities in Hematology and Transfusion medicine. My initial exposure to laboratory research was a yearlong project during Medical School. After training in Pediatrics, I entered the Hematology-Oncology Fellowship at the Children's Hospital of Philadelphia, which I combined with a fellowship in Blood Banking and Transfusion medicine at the Hospital of th University of pennsylvania. My laboratory research was focused on a human model system of cold agglutinin autoimmune hemolytic anemia associated with a B cell lymphoma. The significance of this work lay in the demonstration, based on immunoglobulin gene analysis, that antigen driven clonal selection might play a role in th pathogenesis of the lymphoma. i have since pursued laboratory projects addressing aspects of clonal selection and V gene use in human autoantibodies, such as cold agglutinins, and in B cell neoplasms such as B-CLL and immunodeficiency related lymphomas. i started as Assistant Professor of Pediatrics at Children's Hospital in July 1992. Through this award, I hope to gain the independence and scientific recognition needed to establish a laboratory research program in molecular immunology which will complement and enhance my clinical activities. The proposed research addresses the role of antigen or ligand mediated selection in the formation and expression of the immunoglobulin VH gene repertoire during normal development. The goal is to determine if there is restriction of the VH gene repertoire in normal peripheral B cell and bone marrow pre-B cell compartments, and how such patterns of restriction change with development. The approach will be to prepare unbiased libraries of VH genes from defined B cell populations in umbilical cord blood an later in development, and to evaluate these libraries by a combination of screening with oligonucleotide probes and selective sequencing to identify individual VH gene segments. The results of these studies will have their greatest significance in the interpretation of the apparent VH gene restriction observed in disease states, such as B cell neoplasms and autoimmunity, and perhaps in immunodeficiencies. The sponsor, Dr. leslie Siberstein, is an established investigator in molecular genetic approaches to human autoimmune disease, specifically cold agglutinin disease, and to human B cell neoplasia. His laboratory is an ideal environment in which to pursue the proposed research, receive informed criticism, and ultimately develop an independent research program. The co-sponsor, Dr. Richard Hardy, is a leading figure in B cell immunology whose expertise in B cell development and in flow cytometry will be of enormous benefit to my project. my position at the Children's Hospital of Philadelphia will provide access to necessary clinical specimens and to the resources of the Division of Hematology and the Department of Pathology which have made commitments to support the development of my career as a physician-scientist.

我的长期目标是从事学术儿科职业 结合了血液学研究和临床活动的结合 输血医学。 我最初对实验室研究的接触是 医学院期间长一年的项目。 在儿科训练后，我 进入儿童医院的血液肿瘤学奖学金 费城，我将其与血液银行的研究金相结合， 宾夕法尼亚大学医院的输血医学。 我的 实验室研究的重点是冷凝集素的人类模型系统 与B细胞淋巴瘤相关的自身免疫性溶血性贫血。 这 这项工作的意义在于基于免疫球蛋白的演示 基因分析，抗原驱动的克隆选择可能在 淋巴瘤的发病机理。 从那以后，我从事实验室项目 解决人类克隆选择和V基因使用的各个方面 自身抗体，例如冷凝集素，在B细胞肿瘤中，例如 B-CLL和免疫缺陷相关的淋巴瘤。 我最初是助理 1992年7月，儿童医院儿科教授。 通过这个奖项，我希望获得独立性和科学 建立分子实验室研究计划所需的认可 免疫学将补充和增强我的临床活动。 这 拟议的研究解决了抗原或配体介导的作用 在免疫球蛋白VH基因的形成和表达中的选择 正常发展期间的曲目。 目标是确定是否存在 正常外周B细胞和骨骼中VH基因库的限制 骨髓前B细胞室，以及这种限制模式如何改变 随着发展。 该方法将是准备无偏见的VH库 后来 开发，并通过筛选组合评估这些图书馆 使用寡核苷酸探针和选择性测序以识别个体 VH基因段。 这些研究的结果将拥有最大的 在明显VH基因限制的解释中的重要性 在疾病状态中观察到，例如B细胞肿瘤和自身免疫性，以及 也许是在免疫缺陷中。 赞助商莱斯利·西伯斯坦（Leslie Siberstein）是一位成熟的研究员 人类自身免疫性疾病的分子遗传方法，特别是冷 凝集素疾病和人类B细胞肿瘤。 他的实验室是 追求拟议研究的理想环境，接受 知情的批评，并最终制定了一个独立的研究计划。 共同发起人理查德·哈迪（Richard Hardy）博士是B细胞免疫学领域的主要人物 其在B细胞开发和流式细胞术中的专业知识将为 对我的项目的巨大好处。 我在儿童医院的职位 费城将提供必要的临床标本和 血液学和病理学系的资源 已经做出了支持我职业生涯发展的承诺 医师科学家。