An RA immune system derived from patient's stem cells

描述(申请人提供)：最近在小鼠体内模拟人类免疫系统的研究取得了进展，涉及免疫缺陷小鼠胸腺成熟的人骨髓造血干细胞，接受部分相合的人胎儿胸腺组织，以重建捐赠者个性化的T和B细胞库。这些小鼠的功能和多样性的人类T细胞库在自我和非自我识别的耐受性方面与捐赠者的库相似，但不同的是在发育的早期被重建，主要由幼稚的表型T细胞组成。有趣的是，T1D患者干细胞产生了类似多样化的T细胞谱系，但表现出更成熟的记忆效应表型。这表明干细胞中包含的T1D易感基因促进了自身免疫的发展。我们建议进行一项探索性研究，以检验应用这种方法在小鼠体内重建RA患者在自身免疫和疾病发展之前的T和B细胞库的可行性，这将有助于确定易感基因在淋巴细胞发育异常中的作用，选择和调节RA的发展。这将提供一个基于人类RA免疫系统及其决定基因的RA实验模型，随后可以使用所有的小鼠遗传学和实验免疫学技术来研究该模型，并使用在患者身上不可能进行的干预。在这个初步项目中，我们将确定在疾病发展之前的状态下重建RA患者免疫系统的可行性，并获得关于重组免疫系统的不同免疫学特征随MHC易感基因的贡献以及单个重组RA免疫系统向自身反应性和自身免疫进展的速度和程度的初步数据。在AIM2中，我们探索了佐剂或瓜氨酸蛋白免疫以及培养的成纤维细胞样滑膜衬里细胞接种加速自身免疫症状进展的可能性，并确定了主导供体CD4+CD28零亚型是否在重组免疫系统中重建。

DESCRIPTION (provided by applicant): Recently an advance in modeling the human immune system in mice was reported, involving thymic maturation of human bone marrow hematopoietic stem cells in immunodeficient mice receiving partially HLA-matched human fetal thymic tissues to recreate the donor's individualized functional T and B cell repertoires. The functional and diverse human T cell repertoire of these mice resemble the donor repertoire in tolerance of self and non-self recognition, but is distinguished by being recreated at an earlier period of development, predominantly consisting of naive phenotype T cells. Intriguingly, T1D patient stem cells engender a similarly diverse T cell repertoire, but which exhibits much more maturation towards memory-effector phenotype. This suggests T1D susceptibility genes contained in the stem cells foster the development of autoimmunity. We propose an exploratory study to examine the feasibility of applying this methodology to recreate in the mouse the T and B cell repertoires of RA patients at a stage prior to the development of autoimmunity and disease that will enable determining the role of susceptibility genes on abnormalities in lymphocyte development, selection and regulation underlying the development of RA. This will provide an experimental model of RA based on the human RA immune system and its determinative genes that can be subsequently studied with all of the technology of murine genetics and experimental immunology, and using interventions that would not be possible in a patient. In Aim1 of this preliminary project we will determine the feasibility of reconstituting th RA patient's immune system in a state antecedent to development of disease, and acquire preliminary data on the differing immunologic character of the reconstituted immune systems as a function of the contribution of MHC susceptibility genes and the rate and extent to which the individual reconstituted RA immune systems progress to self-reactivity and autoimmunity. In Aim2 we explore the potential of immunization with adjuvant or citrullinated proteins and administration of cultured fibroblastoid synovial lining cells to accelerate the progression of autoimmune manifestations, and determine whether dominant donor CD4+CD28null subset clonotypes are recreated in the reconstituted immune system.