Mechanisms of T-cell induced-APC cytotoxicity in lupus

描述（由申请人提供）：申请要求资金与主要研究者开发一个独立的研究计划的具体意图。申请人在过去四年中一直在从事T细胞免疫学和狼疮（SLE）发病机制领域的基础科学研究。该提案是申请人目前对单核细胞/巨噬细胞（M θ）凋亡的研究的延伸。假设：凋亡诱导分子介导由CD 4+狼疮T细胞引起的自体单核细胞/M θ杀伤。通过这种机制杀死靶细胞可导致自身抗体的产生。具体目的：确定lupusCD 4 + T细胞诱导单核细胞/M θ细胞凋亡的途径。申请人将测试是否可能通过阻断自身反应性CD 4 + T细胞杀伤单核细胞/M θ的凋亡途径来抑制SLE动物模型中自身免疫的发展。巨噬细胞凋亡在触发或增强自身免疫中的作用也将被研究。方法：a）通过流式细胞术测量SLE和对照T细胞上死亡受体配体的细胞表面表达。B）通过细胞毒性测定，确定这些凋亡途径在SLE单核细胞/M θ中是否有功能，以及阻断这些分子是否可以抑制SLE T细胞对自体单核细胞/M θ的杀伤。c）考虑到参与M θ细胞毒性的途径的冗余，申请人将在体外测试抑制死亡受体（FADD、半胱天冬酶、FLIP）下游的死亡信号是否足以抑制由这些配体诱导的单核细胞/M θ细胞凋亡。d）体内研究将试图表征单克隆抗体或融合蛋白对单核细胞/巨噬细胞死亡受体配体的阻断是否可以抑制鼠SLE的发展，以及组织巨噬细胞的消除本身（在体内用氯膦酸盐脂质体）是否足以在动物模型中诱导自身免疫。提出的研究结果可能会确定参与SLE自身抗原产生的潜在机制。这些可能导致开发旨在逆转这些异常并消除或阻断这种疾病的发作和严重程度的治疗干预措施。赞助商和机构致力于为申请人提供受保护的时间，职业发展和资源。

DESCRIPTION (provided by applicant): The application requests funding with the specific intent of developing an independent research program by the principal investigator. The applicant has been pursuing basic science research in the areas of T cell immunology and pathogenesis of lupus (SLE) for the past four years. The proposal is an extension of the applicant's current research on monocyte/macrophage (M theta) apoptosis. Hypothesis: Apoptosis-inducing molecules mediate the autologous monocyte/M theta killing caused by CD4+ lupus T cells. Target cell killing by this mechanism can lead to the generation of autoantibodies. Specific aims: To determine the pathways involved in monocyte/M theta apoptosis induced by lupusCD4+ T cells. The applicant will test whether it's possible to inhibit the development of autoimmunity in an SLE animal model, by blocking the apoptotic pathways involved in monocyte/M theta killing by autoreactive CD4+ T cells. The role of macrophage apoptosis in triggering or augmenting autoimmunity will also be investigated. Methods: a) Measurement of cell surface expression of death-receptor ligands on SLE and control T cells by flow cytometry. b) With cytotoxicity assays, determine whether these apoptotic pathways are functional in SLE monocytes/M theta and whether blocking these molecules can inhibit the autologous monocyte/M theta killing by SLE T cells. c) Given the redundancy of the pathways involved in M theta cytotoxicity, the applicant will test, in vitro, if inhibiting the death signals downstream of the death receptors (FADD, caspases, FLIP) is sufficient to inhibit monocyte/M theta apoptosis induced by these ligands. d) In vivo studies will try to characterize whether the blockade on monocyte/macrophage death-receptor ligands by monoclonal antibodies or fusion proteins, can inhibit the development of murine SLE, and whether the elimination of tissue macrophages; per se (with clodronate liposomes in vivo) is sufficient to induce autoimmunity in an animal model. The results of the studies proposed might identify potential mechanisms involved in the generation of autoantigens in SLE. These could lead into the development of therapeutic interventions designed to reverse these abnormalities and abrogate or block the onset and severity of this disease. The sponsor and the institution are committed to contributing protected time, career development and resources to the applicant.