Role of CD30 in Alloimmune Responses and Graft Rejection

CD30 在同种免疫反应和移植物排斥中的作用

• 批准号: 6934660
• 负责人: JAY CAMPISI
• 金额: $ 4.07万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2006-05-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6934660
• 关键词: CHO cells; T lymphocyte; apoptosis; biological signal transduction; cytokine receptors; cytotoxicity; enzyme linked immunosorbent assay; flow cytometry; genetically modified animals; heart transplantation; histocompatibility; homologous transplantation; immune response; immune tolerance /unresponsiveness; immunocytochemistry; interferon gamma; interleukin 10; interleukin 13; interleukin 2; interleukin 5; laboratory mouse; leukocyte activation /transformation; lymphocyte proliferation; postdoctoral investigator; protein structure function; transplant rejection; western blottings

DESCRIPTION (provided by applicant): Co-stimulatory molecules such as CD28 and CD40L are necessary for complete T cell activation and play a crucial role in mediating alloimmune responses and allograft rejection. Recently, members of the tumor necrosis factor receptor (TNFR) family have been implicated as additional co-stimulatory molecules that modulate alloimmune responses and graft rejection. CD30 is a pleiotropic TNFR family member that can promote cellular proliferation, cytokine production, and apoptosis depending on the cell type and its state of differentiation. However, little is known about the potential role of CD30+ T cells in alloimmune responses. Therefore, the proposed research plan is designed to test the hypothesis test that CD30 acts as a co-stimulatory molecule in allogeneic responses and modulates T cell proliferation and cytokine production and, subsequently, graft survival. (1) We will determine the effect of CD30 deficiency on T cell function in vivo and in vivo alloreactivity by (a) measuring proliferation, activation, and apoptosis of T cells in CD30-/- and CD30 ++ mice following in vivo and in vitro allostimulation using flow cytometry, CFSE staining, and PE-annexin V staining, (b) measuring T cell cytokine production in CD30-/- and CD30+/+ mice following in vivo and in vitro allostimulation using ELISPOT, ELISA, and RPA and (c) examining the effect of CD30 signal transduction on cell cycle and apoptosis related proteins by RPA and Western blot. (2) We will determine the effect of CD30 deficiency on allograft rejection and on the immune response to the graft by (a) measuring graft survival following vascularized heterotopic cardiac transplants in CD30 and CD30 mice and (b) measuring the function and the phenotype of graft infiltrating cells following allograft transplantation in CD30"/"and CD30 +/+mice using ELISA, cellular proliferation assays, cytotoxicity assays, flow cytometry, H & E staining, and immunohistochemistry. Understanding the contribution of CD30 to alloreactivity will provide novel opportunities to modulate the function of allospecific T lymphocyte populations.

描述（由申请人提供）：共刺激分子如CD28和CD40L是T细胞完全激活所必需的，并且在介导同种免疫反应和同种异体移植排斥中发挥着至关重要的作用。最近，肿瘤坏死因子受体（TNFR）家族的成员被认为是调节同种免疫反应和移植物排斥的额外共刺激分子。 CD30 是一种多效性 TNFR 家族成员，可以根据细胞类型及其分化状态促进细胞增殖、细胞因子产生和凋亡。然而，人们对 CD30+ T 细胞在同种免疫反应中的潜在作用知之甚少。因此，拟议的研究计划旨在测试 CD30 在同种异体反应中作为共刺激分子并调节 T 细胞增殖和细胞因子产生以及随后的移植物存活的假设检验。 (1) 我们将通过 (a) 测量 CD30-/- 和 CD30++ 小鼠体内和体内 T 细胞的增殖、活化和凋亡来确定 CD30 缺陷对体内 T 细胞功能和体内同种异体反应性的影响。使用流式细胞术、CFSE 染色和 PE-annexin V 染色进行体外同种刺激，（b）使用体内和体外同种刺激后测量 CD30-/- 和 CD30+/+ 小鼠中 T 细胞细胞因子的产生ELISPOT、ELISA 和 RPA，(c) 通过 RPA 和 Western blot 检查 CD30 信号转导对细胞周期和凋亡相关蛋白的影响。 (2) 我们将通过以下方式确定 CD30 缺陷对同种异体移植物排斥和移植物免疫反应的影响：(a) 测量 CD30 和 CD30 小鼠血管化异位心脏移植后的移植物存活率，以及 (b) 测量移植物的功能和表型使用 ELISA、细胞增殖测定、细胞毒性测定、流式细胞术、H & E 对 CD30"/" 和 CD30 +/+ 小鼠进行同种异体移植后的移植物浸润细胞染色和免疫组织化学。了解 CD30 对同种异体反应性的贡献将为调节同种异体 T 淋巴细胞群的功能提供新的机会。