A Humanized Mouse Model of B-islet Cell Autoimmunity

描述(申请人提供)：人源化b-胰岛细胞自身免疫小鼠模型。这项建议的目标是在糖尿病允许的小鼠背景上建立一个带有人源化的CD4T细胞室的小鼠模型，在其中研究控制b-胰岛细胞特异性自身免疫的不同方法。由于小鼠和人类不变(Li)物种在伴侣功能上的显著差异，人类白细胞抗原II类转基因小鼠也被提供了人类不变(Li)转基因(Hu-li)和小鼠不变(Li)基因敲除。由于两种小鼠胰岛素原(P-LNS)变种(P-Ins1或P-LNS 2)都不包含以前在人类白细胞抗原DR*0401转基因小鼠和人类白细胞抗原DR*0401阳性T1D患者中发现的主要(C-肽/A链)CD4T细胞表位，因此也增加了正确表达的人胰岛素原转基因基因(Hu-PPI)。现有的具有免疫活性的HLA-DR*0401、DQ8、Hu-CD4、Ab-/-、NOD小鼠正在与新的人源化LI和P-LNS转基因NOD小鼠杂交。为了考察人胰岛素原和胰岛素原物种人源化的效果，以人胰岛素原为自身蛋白的人源化小鼠，将用于研究人P-LNS免疫后的T细胞受体谱和细胞因子反应。我们相信，这些小鼠可能是T1D新的胰岛特异性载体免疫疗法预测试的关键角色。该方案概述了如何在同样携带Rag-/-，GC-/-双重敲除的NOD背景下获得类似的人源化免疫缺陷小鼠模型，在该模型中，研究了组织相容绿色荧光蛋白(GFP)标记的小鼠HSC转移后功能免疫系统的发育、成熟和分布。这些小鼠也将是通过过继转移GFP标记的潜在自体反应性CD4和CDS T细胞群的理想受体，在人源化的T1D小鼠模型中触发疾病。最后，该建议建议使用人源化小鼠模型的免疫缺陷版本来研究“疫苗研究”中诱导主动免疫和耐受的不同条件，其中产生人胰岛素原的树突状细胞(DC)将与不同的CD4和/或CDS T细胞群共同转移到组织相容的小鼠中，小鼠携带糖尿病易感基因HLA-DR*0401，DOS等位基因，人化黎族物种，以及正确表达的人类P-LNS转基因基因。

DESCRIPTION (provided by applicant): A humanized mouse model of b-islet cell autoimmunity. The goal of this proposal is to produce a mouse model on a diabetes permissive murine background with a humanized CD4 T cell compartment, in which to study different approaches towards controlling b-islet cell specific autoimmunity. Due to significant differences in the chaperone function of the murine and human invariant (li) species, the HLA class II transgenic mice, are also being supplied with a human li transgene (hu-li) and a gene knock out for the murine li. Since neither of the two murine Proinsulin (P-lns) varieties (P- Ins1 or P-lns2) contain the major (C-peptide/A-chain) CD4 T cell epitope previously identified in both HLA- DR*0401 transgenic mice and HLA-DR*0401 positive T1D patients, a correctly expressed human preproinsulin transgene (hu-PPI) is also being added. Existing immuno-competent HLA-DR*0401, DQ8, hu- CD4, Ab-/-, NOD mice are being crossed with new humanized li and P-lns transgenic NOD mice. In order to examine the effects of humanizing the li and the Preproinsulin species, the humanized mice, in which human Proinsulin is now a self-protein, will be used to study T cell receptor repertoires and cytokine responses after immunization with human P-lns. We believe that these mice could be key players in pre-testing new islet specific vector based immuno-therapies in T1D. The proposal outlines how to obtain a similarly humanized immuno-deficient mouse model on a NOD background also carrying the Rag-/-, gc-/- double knock out, in which to study the development, maturation, and distribution of a functional immune system after transfer of histo-compatible green fluorescent protein (GFP) labeled murine HSC. These mice will also be ideal recipients for adoptive transfer of GFP labeled potentially autoreactive CD4 & CDS T cell populations triggering disease in the humanized mouse model of T1D. Finally, the proposal suggests to use the immuno- deficient version of the humanized mouse model to examine the different conditions for induction of active immunity versus tolerance in "vaccination studies", where human Proinsulin producing dendritic cells (DC) will be co-transferred with different CD4 and/or CDS T cell populations into histocompatible mice carrying the diabetes susceptible HLA-DR*0401, DOS alleles, a humanized li species, as well as, a correctly expressed human P-lns transgene.