B cell regulation of diabetogenic activity

描述（由申请人提供）：为什么库的特定组分发展或未能发展是一个复杂的问题，与在不同水平上决定免疫优势的力量有关。NOD小鼠具有产生自身免疫性库并屈服于1型糖尿病（T1 D）的倾向。尽管如此，NOD的问题有很多方法可以很容易地得到纠正。我们想探索这些途径中的一些，调节疾病的过程，并涉及B细胞的影响，特别是影响T细胞库的组成。首先，在NOD中自发出现的某些流行克隆型的不同小组将通过使用CDR 3长度光谱法在整个糖尿病前期和糖尿病时期进行跟踪，并且在“特征峰”（SP）出现之后，每个特征峰反映BV/BJ基因家族内CDR 3长度的3000个可能峰之一。是否有一个明确的程序或随机或同时出现的公共克隆型的顺序的问题将被问到。然后将研究B细胞充足的NOD小鼠和B细胞缺乏的NOD小鼠之间SP的差异。将寻求CD 4和CDS克隆外观的一致性，询问其是否依赖于其特定决定簇的接近性。有了克隆型生活史的知识，我们将探讨两个令人烦恼的问题。有数据强调了B细胞作为抗原呈递细胞在NOD中募集致病T细胞的重要性：在B细胞敲除小鼠中，不发生糖尿病，也不能产生某些相关的Ag特异性应答。这表明B细胞和T细胞识别的决定簇之间的关系非常严格，而且树突细胞和巨噬细胞不能产生某些抗原决定簇。我们将用不同的B细胞群体重建B基因敲除小鼠，以尝试了解对特定B细胞的严格要求的规则。活化的B细胞是否比幼稚B细胞更有效可以用转基因B细胞来发现。使用环磷酰胺，其去除或灭活调节细胞，将有可能通过在药物处理的小鼠中观察它们的消失来鉴定特定的SP作为调节细胞。或者，可以披露某些对管制特别敏感的占主导地位的特殊产品。将进行用B细胞Ag-Ig构建体活化抗原特异性T细胞的研究。我们将彻底测试的保护机制是由作为APC的B细胞内的MHC分子捕获决定簇，并且将使用在相同的肽链或基因片段上含有捕获决定簇和待捕获决定簇的嵌合肽。

DESCRIPTION (provided by applicant): Why particular components of the repertoire develop or fail to develop is a complex problem related to the forces that determine immunodominance at various levels. The NOD mouse has a propensity for generating an autoimmune repertoire and succumbing to Type 1 diabetes (T1D). Nevertheless, there are a variety of ways that the NOD's problems can be readily corrected. We would like to explore some of these pathways that modulate the disease process and involve B cell effects that specifically affect the composition of the T cell repertoire. First, a distinct panel of certain prevalent clonotypes that arise spontaneously in the NOD will be followed throughout the prediabetic and diabetic periods, by using CDR3-length spectroscopy, and following the appearance of "signature peaks" (SPs), each reflecting one of the 3000 possible peaks of CDR3 lengths within BV/BJ gene families. The question of whether there is a clear programmed or random or a simultaneous order of appearance of public clonotypes will be asked. The difference in SPs between NOD mice in B cell sufficient and B cell deficient mice will then be studied. The coincidence of CD4 and CDS clonal appearance will be sought, asking whether it is dependent on the proximity of their specific determinants. With the knowledge of the clonotypic life histories, we will approach two vexing problems. There are data emphasizing the importance of the B cell as an antigen presenting cell in enlisting disease- causing T cells in the NOD: in a B cell knockout mouse, diabetes does not occur, nor can certain relevant Ag-specific responses be made. This suggests that there are great stringencies in the relationships between determinants recognized by the B cell and T cell, and also, that dendritic cells and macrophages cannot generate certain antigenic determinants. We will reconstitute B knockout mice with different populations of B cells to attempt to learn the rules for the stringent requirement for particular B cells. Whether activated B cells are more effective than naive B cells can be discovered with transgenic B cells. Using cyclophosphamide, which removes or inactivates regulatory cells, it will be possible to identify particular SPs as regulatory cells, by following their disappearance in drug-treated mice. Alternatively, certain dominant SPs may be disclosed which are particularly sensitive to regulation. Studies of activation of antigen-specific T cells with B cell Ag-lg constructs will be performed. The mechanism of protection that we will test thoroughly is determinant capture by MHC molecules within B cells acting as APC, and chimeric peptides will be employed that contain the capturing determinant and the determinant to be captured on the same peptide chain or gene segment.