Regulation of Gender Dependent EAE Susceptibility

描述(申请人提供)：这项申请探索了在性别和年龄相关的自身免疫性疾病抵抗/易感性模型中CD25+T细胞和NK1.1+细胞对APC活性的动态平衡调节。诱发实验性自身免疫性脑脊髓炎(EAE)的脑源性Th1细胞在所有年龄段的女性和成熟男性(12周龄)中都被激活。相比之下，抗原特异的Th2细胞在年轻成年(6周大)男性中被激活，导致EAE抵抗。去除抗性男性的CD4+CD25+T细胞，而不是年龄匹配的女性，将证明CD25+T细胞抑制Th1的激活。领养转移将证明来自抗性6周男性的CD25+T细胞抑制女性和成熟男性受体的EAE。我们的假设，即APC在体内改变了CD4+CD25+T细胞，将通过诱导来自CD25+耗尽的6周男性供者的APC耐药的6周男性受者的EAE来检验。在NK1.1+细胞耗尽或NK1.1耐药供者的APC转移后，对EAE耐药的男性也能诱导Th1反应。通过阻滞剂或受体配体和对DAP 12KO小鼠的分析，验证了耐药雄鼠NK细胞频率增加通过细胞-细胞相互作用改变APC的假设。通过体内和体外方法研究CD25+T细胞、NK细胞和APC之间的三方相互作用，通过Th2激活导致EAE抵抗。在NK或CD25+T细胞耗尽后，将分析每种细胞类型对其他两种细胞类型细胞因子分泌、表面和基因表达的影响。这些实验将对受性别和年龄调节的单一固定基因内的细胞-细胞相互作用进行机械分析，最终影响对自身免疫性疾病的抵抗力或易感性。这些数据还将提供第一个证据，证明在抗原相遇之前，NK和CD25+T细胞通过改变APC功能来影响适应性免疫。

DESCRIPTION (provided by applicant): This application explores the homeostatic regulation of APC activity by CD25+ T cells and NK1.1+ cells in a gender and age dependent autoimmune disease resistance/susceptibility model using SJL mice in a Th1 mediated CNS autoimmune disease. Encephalitogenic Th1 cells inducing experimental autoimmune encephalomyelitis (EAE) are activated in females of all ages and mature males (>12 wks old). By contrast, Ag specific Th2 cells are activated in young adult (6 wk old) males, resulting in EAE resistance. Depletion of CD4 + CD25 + T cells from resistant males, but not age matched females, will demonstrate that CD25+T cells suppress Th1 activation. Adoptive transfers will demonstrate that CD25 + T cells from resistant 6 wk males inhibits EAE in female and mature male recipients. Our hypothesis, that the APC alters the CD4 + CD25 + T cells in vivo will be tested by inducing EAE in resistant 6 wk male recipients of APC derived from CD25 + depleted 6 wk male donors. Th1 responses are also induced in EAE resistant males either depleted of NK1.1 + cells or following APC transfer from NK1.1 depleted resistant donors. The hypothesis that the increased frequency of NK cells in resistant males alters the APC via cell-cell interaction is examined by blockade or receptor ligand and analysis of DAP 12 KO mice. Tripartite interactions between CD25 + T cells, NK cells and APC which lead to EAE resistance via Th2 activation are examined by in vivo and in vitro approaches. The influence of each cell type on cytokine secretion, surface and gene expression in the other two cell types will be analyzed following depletion of either the NK or CD25 + T cells. These experiments will provide a mechanistic analysis of cell-cell interactions within a single fixed genotype regulated by both gender and age which ultimately influence resistance or susceptibility to autoimmune disease. These data will also provide the first evidence that NK and CD25 + T cells influence adaptive immunity prior to antigen encounter via alterations in APC function.