ANTIGENIC SPECIFICITY AND V REGION USE OF AIDS B NHLS

AIDS B NHLS 的抗原特异性和 V 区使用

• 批准号: 3199266
• 负责人: MICHAEL Shannon MCGRATH
• 金额: $ 18.46万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-04-01 至 1995-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3199266
• 关键词: AIDS; B lymphocyte; Epstein Barr virus; antigen antibody reaction; biopsy; cell bank /registry; clone cells; cytomegalovirus; gene rearrangement; human subject; hybridomas; immunoglobulin genes; immunoglobulin idiotypes; lymphoma; molecular cloning; monoclonal antibody; neoplasm /cancer genetics; neoplastic cell; polymerase chain reaction; virus antigen; virus related neoplasm /cancer

The overall goal of this proposal is to test whether AIDS-associated B-cell non-Hodgkin's lymphomas (NHL) represent outgrowths of viral or self antigen activated B-cells, to define the spectrum of immunoglobulin variable (V) region genes utilized by these lymphomas and to test whether subsets of lymphoma would utilize identical V regions. If viral antigens are found to drive the pre-malignant B-cell proliferation or common V region determinants are shared between different lymphomas, non-immunosuppressive therapeutic approaches to the prevention and treatment of AIDS-lymphomas could be devised. In a preliminary analysis of two AIDS lymphoma cell lines we found one AIDS NHL IgM to be HIV-1 reactive (2F7) and the other to be self reactive (10C9). Rather than laboriously and inefficiently establish other AIDS NHL cell lines, we will create AIDS NHL hybridomas for testing of IgM antigenic specificity. In order to rapidly clone V regions and compare V region sequences between large numbers of patient specimens, a recent advance in polymerase chain reaction (PCR) technology will allow the rapid and direct cloning of rearranged V regions from tumor genomic DNA to test whether, as in other classes of lymphoid malignancy, subsets of AIDS NHL will be found to utilize common V regions. Specific aims: 1) To test IgM's from monoclonal and polyclonal AIDS-associated lymphomas for reactivity with a panel of viral (HIV-1, EBV, CMV) and self antigens, and utilizing polymerase chain reaction (PCR), to clone the IgM variable region genes from at least ten monoclonal tumors in order to define the spectrum of V region genes (and V region modifications) used by these lymphomas. 2) To characterize the molecular nature of the monoclonal idiotype shared between the 10C9 self reactive lymphoma cell line and 5 of 21 monoclonal AIDS-associated lymphomas. 3) To use specific AIDS lymphoma V region (CDR2) oligonucleotides from the ten cloned VH regions including the 10C9 V region for analysis of PCR amplified VH genes from frozen and formalin fixed archival AIDS NHL specimens (currently greater than 125 at SFGH) to determine the prevalence of specific VH region utilization in AIDS-associated lymphomas.

这项提案的总体目标是测试艾滋病相关的B细胞是否 非霍奇金淋巴瘤（NHL）代表病毒或自身抗原的副产物 激活的B细胞，以确定免疫球蛋白可变区（V）的谱 区域基因利用这些淋巴瘤，并测试是否子集 淋巴瘤会利用相同的V区 如果发现病毒抗原 驱动癌前B细胞增殖或共同V区 决定簇在不同的淋巴瘤之间共享，非免疫抑制 预防和治疗艾滋病淋巴瘤的治疗方法 可以设计。 在两个艾滋病淋巴瘤细胞系的初步分析中，我们发现一个艾滋病 NHL IgM为HIV-1反应性（2F 7），另一种为自身反应性 （10C9）。 而不是费力和低效地建立其他艾滋病NHL 细胞系，我们将建立艾滋病非霍奇金淋巴瘤杂交瘤检测IgM抗原 的特异性 为了快速克隆V区并比较V区 大量患者标本之间的序列，最近的进展， 聚合酶链反应（PCR）技术将允许快速和直接 从肿瘤基因组DNA中克隆重排的V区，以测试是否， 在其他类型的淋巴系统恶性肿瘤中，会发现AIDS NHL的亚类， 以利用公共V区。 具体目标： 1)检测单克隆和多克隆艾滋病相关淋巴瘤的IgM 对于与一组病毒（HIV-1、EBV、CMV）和自身抗原的反应性， 并利用聚合酶链反应（PCR）克隆IgM可变区 区域基因从至少10个单克隆肿瘤，以确定 它们使用的V区基因（和V区修饰）的谱 淋巴瘤 2)表征共享的单克隆独特型的分子性质 10 C9自身反应性淋巴瘤细胞系与21个单克隆抗体中的5个之间存在显著性差异（P <0.05）。 艾滋病相关淋巴瘤 3)为了使用特异性AIDS淋巴瘤V区（CDR 2）寡核苷酸， 10个克隆的VH区，包括10 C9 V区，用于PCR分析 从冷冻和福尔马林固定的档案AIDS NHL扩增VH基因 样本（目前在SFGH超过125），以确定患病率 艾滋病相关淋巴瘤中特异性VH区的利用。