A Humanized Mouse Model of B-islet Cell Autoimmunity

B 胰岛细胞自身免疫的人源化小鼠模型

• 批准号: 7623861
• 负责人: GRETE SONDERSTRUP
• 金额: $ 30.94万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7623861
• 关键词: Active immunity; Adoptive Transfer; Alleles; Architecture; Autoimmunity; C-Peptide; CD4 Positive T Lymphocytes; Canada; Cells; Collaborations; Commit; DNA; Dendritic Cells; Development; Diabetes Mellitus; Disease; Funding; Genes; Genomics; Goals; Green Fluorescent Proteins; Homing; Human; Human Development; IL2RA gene; Immune system; Immunization; In Vitro; Inbred NOD Mice; Insulin; Insulin-Dependent Diabetes Mellitus; Islet Cell; Knock-out; Knockout Mice; Label; Laboratories; Lymphoid; Lymphoid Cell; MHC Class I Genes; Molecular Chaperones; Mus; Nucleic Acid Regulatory Sequences; Organ; Pathway interactions; Patients; Population; Procedures; Proinsulin; Property; Proteins; Relative (related person); Research Personnel; System; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Epitopes; TNFRSF10A gene; Testing; Transgenes; Transgenic Mice; Transgenic Organisms; Universities; Vaccination; base; cytokine; hybridoma production; improved; invariant chain; islet; mouse model; peptide A; preproinsulin; prevent; programs; research study; response; selective expression; vector

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.

描述（由申请人提供）：b-胰岛细胞自身免疫的人源化小鼠模型。本提案的目标是在糖尿病允许的鼠背景上产生具有人源化CD 4 T细胞区室的小鼠模型，其中研究控制b-胰岛细胞特异性自身免疫的不同方法。由于鼠和人不变（li）物种的伴侣蛋白功能的显著差异，还向HLA II类转基因小鼠提供人li转基因（hu-li）和鼠li的基因敲除。由于两种鼠胰岛素原（P-Ins）品种（P-Ins 1或P-Ins 2）均不含有先前在HLA-DR *0401转基因小鼠和HLA-DR*0401阳性T1 D患者中鉴定的主要（C肽/A链）CD 4 T细胞表位，因此还添加了正确表达的人前胰岛素原转基因（hu-PPI）。现有的免疫活性HLA-DR*0401、DQ 8、hu-⑶ 4、Ab-/-、NOD小鼠与新的人源化li和P-Ins转基因NOD小鼠杂交。为了检查人源化Ii和前胰岛素原种类的效果，将使用人源化小鼠（其中人胰岛素原现在是自身蛋白）来研究用人P-Ins免疫后的T细胞受体库和细胞因子应答。我们相信，这些小鼠可能是T1 D中基于胰岛特异性载体的免疫疗法的关键参与者。该提案概述了如何获得一个类似的人源化免疫缺陷小鼠模型的NOD背景上也进行Rag-/-，gc-/-双敲除，在其中研究的发展，成熟，和功能性免疫系统的分布后，转移组织相容的绿色荧光蛋白（GFP）标记的小鼠HSC。这些小鼠也将是用于在TlD的人源化小鼠模型中过继转移GFP标记的潜在自身反应性CD 4 & CD 8 T细胞群体的理想受体，所述细胞群体触发疾病。最后，该提案建议使用免疫缺陷型的人源化小鼠模型来检查“疫苗接种研究”中诱导主动免疫与耐受的不同条件，其中将人胰岛素原产生树突细胞（DC）与不同的CD 4和/或CDS T细胞群共转移到携带糖尿病易感性HLA-DR*0401，DOS等位基因，人源化Ii物种，以及正确表达的人P-Ins转基因。