ANTIGENIC SPECIFICITY AND V REGION USE OF AIDS B NHLS

这项提案的总体目标是测试与艾滋病相关的B细胞 非霍奇金淋巴瘤(NHL)代表病毒或自身抗原的突起 激活的B细胞，以确定免疫球蛋白可变(V)的谱 这些淋巴瘤所利用的区域基因，并测试其亚群 淋巴瘤会利用相同的V区。如果病毒抗原被发现 驱动癌前B细胞增殖或常见的V区 决定因素在不同的淋巴瘤之间共享，非免疫抑制 防治艾滋病淋巴瘤的治疗方法 是可以设计出来的。 在对两个艾滋病淋巴瘤细胞株的初步分析中，我们发现了一个艾滋病 非霍奇金淋巴瘤IgM为HIV-1反应性(2F7)，另一例为自身反应性 (10C9)。而不是费力和低效地建立其他艾滋病非霍奇金淋巴瘤 细胞系，我们将创建艾滋病NHL杂交瘤用于检测IgM抗原 专一性。为了快速克隆V区并比较V区 大量患者样本之间的序列，最新进展 聚合酶链式反应(PCR)技术将使快速和直接 从肿瘤基因组DNA中克隆重排的V区以测试AS 在其他类型的淋巴系统恶性肿瘤中，会发现艾滋病非霍奇金淋巴瘤的亚型。 利用共同的V区。具体目标： 1)检测与艾滋病相关的单克隆性和多克隆性淋巴瘤的IgM 对于与一组病毒(HIV-1、EBV、CMV)和自身抗原的反应， 利用聚合酶链式反应(聚合酶链式反应)，克隆IgM变量 从至少10个单抗肿瘤中提取区域基因，以确定 它们使用的V区基因(和V区修饰)的谱 淋巴瘤。 2)鉴定共有的单克隆独特型的分子性质 10C9自身反应性淋巴瘤细胞株与21株单抗中5株的比较 与艾滋病相关的淋巴瘤。 3)使用艾滋病淋巴瘤V区(CDR2)的特异性寡核苷酸 包括10C9 V区在内的10个克隆的VH区用于PCR分析 从冰冻和福尔马林固定的艾滋病非霍奇金淋巴瘤中扩增VH基因 样本(目前在SFGH大于125)以确定流行率 艾滋病相关淋巴瘤中特定VH区利用的研究。

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.