B cell regulation of diabetogenic activity

描述（由申请人提供）：为什么抗原库的特定成分发展或不发展是一个复杂的问题，与决定不同水平免疫优势的力量有关。NOD小鼠具有产生自身免疫库和屈服于1型糖尿病（T1D）的倾向。尽管如此，NOD的问题还是有多种容易纠正的方法。我们希望探索其中一些调节疾病过程的途径，并涉及特异性影响T细胞库组成的B细胞效应。首先，在整个糖尿病前期和糖尿病期间，通过使用CDR3长度光谱，跟踪出现的“特征峰”（SPs），每个峰反映BV/BJ基因家族中3000个可能的CDR3长度峰中的一个，跟踪NOD中自发出现的特定流行克隆型的独特面板。问题是是否有一个明确的程序或随机或同时出现的顺序出现的公共克隆型。B细胞充足和B细胞缺乏小鼠NOD小鼠之间SPs的差异将被进一步研究。将寻求CD4和CDS克隆外观的巧合，询问它是否依赖于其特定决定因素的接近性。有了克隆型生命史的知识，我们将探讨两个令人烦恼的问题。有数据强调了B细胞作为抗原提呈细胞在NOD中征召引起疾病的T细胞中的重要性：在B细胞敲除小鼠中，不会发生糖尿病，也不会产生某些相关的ag特异性反应。这表明B细胞和T细胞识别的决定因子之间存在很大的相关性，并且树突状细胞和巨噬细胞不能产生某些抗原决定因子。我们将用不同的B细胞群重组B基因敲除小鼠，试图了解对特定B细胞严格要求的规则。激活的B细胞是否比未激活的B细胞更有效，可以用转基因B细胞来发现。使用环磷酰胺，去除或失活调节细胞，将有可能识别特定的SPs作为调节细胞，通过跟踪它们在药物治疗小鼠中的消失。或者，某些对监管特别敏感的主导SPs可能会被披露。研究活化抗原特异性T细胞与B细胞Ag-lg结构将进行。我们将彻底测试的保护机制是由B细胞内的MHC分子作为APC捕获决定因素，并且将使用嵌合肽，其中包含捕获决定因素和在同一肽链或基因片段上被捕获的决定因素。

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.