LOCAL CYTOKINE ENVIRONMENT IN ACUTE REJECTION

这项研究是基于同种异体移植排斥反应的假设， 是一种抗原依赖性的破坏性炎症过程， 叠加在修复导向的炎症过程中， 同种异体移植物和同种异体移植物。 两个移植物都获得了炎性 细胞浸润，但引发同种异体移植物炎症的信号 不知道。 急性排斥反应，即使逆转，也与 移植物存活率降低， 移植物排斥反应的早期事件对治疗有重要意义 值 我们将通过T淋巴细胞亚群研究早期细胞因子的表达 以及抗原依赖性和抗原非依赖性 内皮炎性粘附分子在小鼠心脏排斥反应中作用 同种异体移植物，并将它们与同种异体I类或I类 II MHC。 具体来说，我们将1）分析T细胞中的细胞因子表达 内皮细胞和内皮细胞中的细胞表面表型 心脏移植排斥反应 我们将使用PCR，免疫组织化学 通过细胞分离的T细胞和内皮细胞的ELISPOT分析 分选以建立细胞因子合成和表达的基线， 内皮细胞表面分子在早期同种异体移植物炎症中的作用。 2)确定CD4+和CD8+ T淋巴细胞各自的贡献 细胞因子在移植物中的表达。 我们将T细胞分离成CD4+ 和CD8+亚群，以确定它们是否贡献不同的细胞因子 以及它们是否被募集到应答中 不同的动力学。 3)确定单个MHC差异的影响 对移植物发育过程中CD4+、CD8+细胞活性的影响 炎症 我们将通过阻止免疫系统的 通过I类或II类MHC激活，并检查对细胞的影响。 募集CD4+和CD8+细胞进入炎症反应，早期 细胞因子的合成和炎症粘附分子的表达， 移植物内皮 4)确定体内干扰的影响 CD4+和CD8+细胞功能对I类移植物炎症的影响， II类异种移植物。 我们将使用针对CD4、CD8和 IFN γ去除或抑制CD4+和CD8+细胞的功能， I类或II类不同移植物。 实际上，我们将剖析 体内同种异体移植物反应，以确定CD4+和CD8+细胞在 抗原依赖性破坏性炎症的发展。 在 总的来说，这些研究将证明T淋巴细胞亚群的作用， 在体内建立同种异体反应的细胞因子环境中。 在这样做的时候，他们将提供有关早期事件的新信息， 同种异体移植物炎症

The proposed studies are based on the hypothesis that allograft rejection is an antigen-dependent, destructive inflammatory process that is superimposed on the repair-oriented inflammatory process that occurs in both the isograft and allograft. Both grafts acquire an inflammatory cellular infiltrate, but the signals that initiate allograft inflammation are not known. Acute rejection, even when reversed, is associated with decreased graft survival, and the identification and inhibition of very early events in graft rejection should be of significant therapeutic value. We will study early cytokine expression by T lymphocyte subsets and the expression of antigen-dependent and antigen-independent inflammatory adhesion molecules by endothelia in rejecting murine cardiac allografts and relate them to the presence of allogeneic class I or class II MHC. Specifically, we will 1) analyze cytokine expression in T cells and endothelial cells and cell surface phenotype in endothelial cells from rejecting cardiac allografts. We will use PCR, immunohistochemistry and ELISPOT analysis of T cells and endothelial cells isolated by cell sorting to establish a baseline of cytokine synthesis and expression of endothelial cell surface molecules during early allograft inflammation. 2) determine the respective contributions of CD4+ and CD8+ T lymphocytes to cytokine expression in the graft. We will separate T cells into CD4+ and CD8+ subsets to determine whether they contribute different cytokines to the alloresponse, and whether they are recruited into the response with different kinetics. 3) determine the effect of single MHC disparity on the activity of CD4+ and CD8+ cells in the development of allograft inflammation. We will perturb the allograft system by disallowing immune activation via class I or class II MHC, and examine the effects on the recruitment of CD4+ and CD8+ cells into the inflammatory response, early cytokine synthesis, and expression of inflammatory adhesion molecules by graft endothelia. 4) determine the effects of interference with in vivo CD4+ and CD8+ cell function on allograft inflammation in class I and class II disparate grafts. We will use antibodies to CD4, CD8 and IFNgamma to remove or inhibit the function of CD4+ and CD8+ cells in class I or class II disparate grafts. In effect, we will dissect the in vivo allograft response to determine the role of CD4+ and CD8+ cells in the development of antigen-dependent destructive inflammation. In general, these studies will demonstrate the role of T lymphocyte subsets in establishing the cytokine environment of the allo-response in vivo. In so doing, they will provide new information regarding early events in allograft inflammation.