Immunoregulation of Autoimmunity

在这项竞争性续展申请中提出的研究意在确定使 (1)在有或没有选择TLR参与的情况下系统地给予抗CD3抗体或(2) 产生或过继转移调节性T细胞(Tregs)，以阻止进展为糖尿病前期 糖尿病或新近发病的高血糖NOD小鼠。正在研究的假设是，这些建议 各种疗法都有一条共同的最终有效途径。 这项提议中的机制研究将使用cDNA微阵列和新的蛋白质表达方法 在核心B中可用，用于比较在治疗小鼠中看到的基因和蛋白质表达模式 正常的结节疾病进展与非炎症的NOD.810组织中看到的模式相比(NOD 之前U19合同中的路线图数据)，以确定观察到的治疗效果的机制(S)。 下列基因和蛋白质表达模式的鉴定、分析和表征 NOD小鼠的成功治疗应对其作用机制、识别 替代标记物的效果，并可能提供额外的或替代的治疗靶点。 根据我们在之前的U19资助下获得的初步数据，出现了两个主要现象 成功治疗新近高血糖的NOD小鼠后；(1)免疫反应的变化 胰岛浸润性T细胞的分布导致基因和蛋白质表达模式的重组 浸润性T细胞的结构，与抑制自身免疫破坏β相一致 细胞，和(2)胰腺β细胞再生(伤口愈合)，作为这一变化的结果。四 已经提出了具体的目标来检验以下假设：注射受内毒素污染的药物 抗CD3抗体或过继细胞疗法，使用Tregs或局部产生的TGFp，直接作用于 自激活淋巴细胞存在于胰岛和引流淋巴结中，或者，正如我们所相信的，通过激活 Tregs直接或继发性地阻断胰岛β细胞的破坏，促进胰岛β细胞 再生作为伤口愈合的一种形式。 这些研究对目前人类新发糖尿病的一种治疗方法有直接影响，即使用 抗CD3抗体。尽管对新近发病的高血糖NOD小鼠的治疗非常成功，但 在人体试验中，抗CD3抗体的反应并不引人注目。如果我们的初选 数据是正确的，在小鼠模型中使用的抗CD3抗体中的污染内毒素 协同，使用TLR激动剂与无内毒素的抗人CDS抗体协同作用的概念 可能在治疗新近发病或糖尿病前期人类疾病方面提供重大的治疗进展。

Studies proposed in this competing renewalapplication intend to identify the mechanisms of effect that allow (1) systemic administration of anti-CD3 antibody with or without selected TLR engagement or (2) the generation of, or adoptive transfer of, regulatory T cells (Tregs), to block progression to diabetes in pre- diabetic or recent onset hyperglycemic NOD mice. The hypothesis being studied is that these proposed therapies share a final common pathway ofeffect. Mechanistic studies in this proposal will use cDNA microarray and novel methods of protein expression available in Core B, to compare gene and protein expression patterns seen in treated mice to those seen in normal NOD disease progression when comparedto patterns seen in non-inflamed NOD.810 tissues (NOD Roadmap data from previous U19 contract),to identify the mechanism(s)of the observedtherapeutic effect. Identification, analysis and characterization of gene and protein expression patterns seen following successful therapy of NOD mice should provide important insights into mechanism of effect, identify surrogate markersof effect, and potentially provide additional or alternative targets for therapy. According to our preliminary data obtained under previous U19 funding, two major phenomenon occur following successful therapy of recently hyperglycemic NOD mice; (1) a change in the immune response profile of islet-infiltrating T cells leading to a reorganization of the gene and protein expression patterns and the architecture of the infiltrating T cells, co-incident with inhibition of the autoimmune destruction of beta cells, and (2) pancreatic beta cell regeneration (wound healing) as a consequence of this change. Four Specific Aims have been proposed to test the following hypothesis: administration of endotoxin contaminated anti-CD3 antibody or adoptive cellular therapy, using Tregs or local TGFp production, acting directly on autpreactive lymphocytes present in the islets and draining lymph nodes, or, as we believe, by activating Tregs, either directly, or secondarily blocks islet beta cell destruction and facilitates islet beta cell regeneration as a form of wound healing. These studies have direct impact on one current therapy of recent onset diabetes in man, the use of anti-CD3 antibodies. Despite extremely successful treatment of recent onset hyperglycemic NOD mice with anti-CD3 antibodies, responses seen in the human trials have been less than spectacular. If our preliminary data are correct, that contaminant endotoxin in the anti-CD3 antibodies used in the mouse model was synergistic, the concept of using TLR agonists in synergy with endotoxin free anti-human-CDS antibodies may provide a major therapeutic advance in the treatment of recent onset or pre-diabetic human disease.