INDUCTION OF AUTOIMMUNITY WITH IDIOTYPES

我们开发了一种新的系统性红斑狼疮实验模型。 其中针对70K U1 RNP核自身抗原的自身抗体， 双链DNA与免疫沉淀性肾小球肾炎 用免疫球蛋白轻链免疫BALB/c小鼠。这盏灯 链属于一组MRL-LPR/LPR单抗(Ab-2) 抗68KU1 RNP多肽MRL-LPR/LPR单抗识别 28/12(抗体-1)。单抗28/12本身具有独特型标志 MRL-LPR/LPR抗DNA抗体。抗体-2的VL区的序列为 与U1 RNP抗原区的氨基酸序列同源 多肽。我们的首要任务是定义 ID-28/12网络。我们已经开始全面审查 具有代表性的Ab2‘S的结构。我们计划建立一个 这些Ab2‘与S的氨基酸序列之间存在相关性 它们能够在BALB/c小鼠体内激发自身抗体的产生。 与70D RNP自身抗原的抗原区的任何关系都将是 特别有趣的事。我们会采用三大策略：(A) C DNA克隆测序；(B)免疫球蛋白的体内检测 序列和定量表征的免疫化学和 独特型特性；和(C)合成多肽免疫，其 氨基酸序列对应于CDR3轻链G.In的氨基酸序列 在进行这些结构研究的同时，我们正在进行一项调查 ID-28/12网络的血清学、独特型和免疫病理学 模特。该项目的这一部分包括免疫化学和独特型 血清、单抗及免疫病理学分析 对有关机关进行调查。免疫小鼠获得杂交瘤 用ID-LGG也将为测序猪的V基因提供材料 引发了自身抗体。对这些序列的确定将测试 假设独特型可以是驱动 克隆性自身免疫反应。我们还计划调查 确定对ID-LCG的自身免疫反应是否受MHC限制。进一步 对自发ID-28/12网络的研究将在 Mrl-+/+小鼠。我们将研究抗体-1随着时间的发展和 AB-2个网络组件(即，哪个组件最先出现？)，并分析 年轻和老年MRL-+/+小鼠中单抗的出现频率 VL CDR3‘S与自身抗原样本型CDR3相关。我们会 也要检验这样的假设，即对沙门氏菌的免疫反应 脂多糖可以触发相应抗体的产生 到ID-28/12网络的Ab-2。我们将获得的信息将 用于开发新的治疗策略，特别是 强调诱导(或恢复)免疫的可能性 自身抗原模拟的合成肽的耐受性 抗体-2的表位。

We have developed a new experimental model of systemic lupus erythematosus in which autoantibodies against the 70K U1 RNP nuclear autoantigen, double-strand DNA and immune-deposit glomerulonephritis can be induced in BALB/c mice by immunization with an immunoglobulin light chain. This light chain belongs to a set of MRL-lpr/lpr monoclonal antibodies (Ab-2) recognized by monoclonal anti-68KU1 RNP polypeptide MRL-lpr/lpr antibody 28/12 (Ab-1). Monoclonal antibody 28/12 itself has an idiotypic marker of MRL-lpr/lpr anti-DNA antibodies. A sequence in the VL region of Ab-2 is homologous to an amino acid sequence in an antigenic region of the U1 RNP polypeptide. Our first priority is to define the structural basis of the Id-28/12 network. We have already begun a comprehensive examination of the structure of representative Ab-2's. We plan to establish whether a correlation exists between the amino acid sequences of these Ab-2's and their ability to evoke the production of autoantibodies in BALB/c mice. Any relationship to the antigenic region of the 70D RNP autoantigen will be of particular interest. Three major strategies will be employed: (a) sequencing of cDNA clones; (b) in vivo assays of immunoglobulins with known sequences and with quantitatively characterized immunochemical and idiotypic properties; and (c) immunization with synthetic peptides whose amino acid sequences correspond to those of the CDR3 light chain G. In parallel with these structural studies we are conducting an investigation of the serology, idiotypes, and immunopathology of the Id-28/12 network model. This part of the project, entails immunochemical and idiotypic analyses of serum and monoclonal antibodies as well as immunopathological investigations of relevant organs. Hybridomas obtained from mice immunized with Id-LGG will also provide material for sequencing the V genes of the evoked autoantibodies. Determination of these sequences will test the hypothesis that an idiotype can be the selective pressure driving a clonally related autoimmune response. We also plan investigations to determine if autoimmune responses to Id-LCG are MHC-restricted. Further studies of the spontaneous Id-28/12 network will be conducted in the MRL-+/+ mouse. We will examine the development over time of the Ab-1 and Ab-2 components of the network (i.e. which one appears first?), and analyze the frequency in young and old MRL-+/+ mice of monoclonal antibodies with VL CDR 3's related to the prototypic autoantigen-mimicking CDR3. We will also test the hypothesis that an immune response to Salmonella RE lipopolysaccharide can trigger the production of antibodies corresponding to the Ab-2 of the Id-28/12 network. The information we will obtain will be used for the development of new therapeutic strategies, with particular emphasis on the possibility of inducing (or restoring) immunological tolerance with synthetic peptides corresponding to autoantigen-mimicking epitopes of Ab-2.