Molecular Basis of Regulatory Idiotopes

以往的研究已经定义了一类独特型的调节独特型，这些独特型在免疫应答的独特型调节中发挥重要作用。具有不同抗原特异性的B细胞克隆之间的高连通性已经被证明，并且这些克隆可以被特定于调节独特型的抗体激活。这种现象被称为“交叉监管”。此外，大多数表达A48调控独特型的单克隆抗体利用来自VH441-4和VK10家族的V基因。进一步的研究将旨在利用这些观察结果来进一步了解调控独特体表达的细胞和分子机制。用VH441种系基因探针从未免疫小鼠和杂交瘤中筛选，制备了A48抗体重链CDR1合成肽的兔抗体。合成肽的抗体将用于确定共享调节独特型的抗体是否与衍生A48蛋白的种系基因表达相同的CDR1序列。从未免疫小鼠（“稳定状态”）中选择的杂交瘤产生的抗体将根据其VH和VK家族，其独特型和精细特异性进行表征。这将使在稳定状态下确定调控独特型的频率成为可能。VH和VK基因编码表达调控独特型的抗体的序列将进行，并将加强我们对促进调控独特型表达的分子因素的理解。这些研究的总体目标是阐明调控异位的免疫化学和分子基础。这一知识应该增加我们对免疫反应调节机制的理解。

Previous studies have led to the definition of a category of idiotope designated regulatory idiotopes which have been proposed to play an important role in idiotype-determined regulation of the immune response. High connectivity between B cell clones with various antigen specificities has been demonstrated, and such clones can be activated by antibodies specific for regulatory idiotopes. This phenomenon has been termed "cross-regulation". Furthermore, the majority of monoclonal antibodies expressing A48 regulatory idiotopes utilized V genes derived from the VH441-4 and VK10 families. Further studies will be aimed at exploiting these observations to gain further insight into the cellular and molecular mechanisms governing the expression of regulatory idiotopes. A rabbit antibody specific for a synthetic peptide corresponding to CDR1 of the heavy chain of A48 antibody and hybridomas from non- immunized mice selected with a VH441 germline gene probe have been prepared. The antibody to the synthetic peptide will be used to determine whether or not the antibodies sharing regulatory idiotopes express the same CDR1 sequence as the germline gene from which the A48 protein is derived. The antibodies produced by the hybridomas selected from non-immunized mice ("steady state") will be characterized with respect to their VH and VK families, their idiotypy, and their fine specificities. This should make it possible to determine the frequency of regulatory idiotopes in the steady state. The sequence of VH and VK genes encoding antibodies expressing regulatory idiotopes will be carried out and should enhance our understanding of the molecular factors contributing to the expression of regulatory idiotopes. The overall goal of these studies is to elucidate the immunochemical and molecular basis of regulatory iditopes. This knowledge should add to our understanding of the mechanisms by which immune responses are regulated.