TRANSPLANT IMMUNOLOGY OF IL2 IFNY DOUBLE KNOCKOUT MICE

IL2 IFNY 双敲除小鼠的移植免疫学

• 批准号: 6372640
• 负责人: Martin S Zand
• 金额: $ 12.04万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6372640
• 关键词: T lymphocyte; blocking antibody; cellular immunity; gene targeting; genetically modified animals; humoral immunity; immune tolerance /unresponsiveness; interferon gamma; interleukin 12; interleukin 15; interleukin 2; interleukin 4; laboratory mouse; pancreas transplantation; secretion; transplant rejection; transplantation immunology

Approximately, 10,000 renal and 1,500 pancreas transplants (allografts) are performed annually in the United States. Despite aggressive immune suppression, hundreds of these organs are lost each year to immune mediated rejection. Because the number of individuals waiting for a transplant greatly exceeds the supply of donated organs, transplant rejection directly contributes to the shortage of these organs. Intragraft expression of the cytokines interleukin 2 and interferon gamma (IFN-gamma) has been found to be markers for clinical and experimental allograft rejection. Although most clinical protocols for immune suppression in transplant patients can reduce the levels of IL-2 and IFN-gamma, immune mediated rejection still occurs. Currently, no experimental models exist in which allograft rejection can be studied when both IL-2 and IFN-gamma are completely absent. The goal of this proposal is to create a transgenic mouse model with targeted gene disruption of both the IL-2 and IFN-gamma genes, and to study the immune responses to transplanted issue in these mice. We have successfully bred IL-2 IFN-gamma double knockout mice (DKO). The mice are generally healthy, have normal litters, and survive as long as 6 months in sterile housing. Our preliminary data indicate that the DKO mice develop a much milder form of the autoimmune colitis seen in the IL-2 single knockout mice. DKO mice can mount a strong delayed-type hypersensitivity. response to a foreign protein antigen. Finally, the DKO mice have a profound defect in B cell maturation. Our specific goals in this proposal are to (1) characterize the ability of the DKO mice to mount cellular and humoral immune responses and (2) determine the outcome of the immune response to pancreatic allografts in the absence of IL-2 and IFN-gamma (i.e. rejection or tolerance). First, we will study the ability of DKO mice to generate delayed -type hypersensitivity responses to histocompatibility antigens (MHC), generate antibodies after immunization, and generate cytotoxic T-lymphocyte. Second, we will render DKO mice diabetic using streptozotocin, and then perform pancreatic islet cell transplantation from MHC disparate donors. Allograft survival will be monitored we hypothesize that the DKO mice will reject islet allografts, and that the mechanisms of rejection will involved IL-4 or other T cell growth factors and cytotoxic T cells. The cytokine milieu and cell phenotypes involved will be identified by quantitative polymerase chain reaction and fluorescence activated cell sorting. In contrast, if the DKO mice are tolerant to islet allografts, we will determine the mechanism by adoptive transfer of tolerance inducing T lymphocyte subsets.

大约10,000例肾移植和1,500例胰腺移植(同种异体移植) 在美国每年都有演出。尽管有攻击性的免疫 抑制，每年有数百个这样的器官失去免疫 中介性排斥。因为有多少人在等待 移植大大超过了捐赠器官的供应，移植 排斥反应直接导致这些器官的短缺。移植物内 细胞因子白介素2和干扰素-γ的表达 已被发现是临床和实验同种异体移植的标志物 拒绝。尽管大多数临床上用于免疫抑制的方案 移植患者可降低IL-2和干扰素-γ水平，免疫 介导性排斥仍然会发生。目前，尚无实验模型。 当IL-2和干扰素-γ同时存在时，可以研究同种异体移植排斥反应 完全不存在。这项提议的目标是创造一种转基因 白介素2和干扰素-γ同时靶向基因干扰的小鼠模型 基因，并研究对移植问题的免疫反应 老鼠。 我们已成功培育出IL-2、干扰素-γ双基因敲除小鼠(DKO)。这个 小鼠一般都是健康的，产仔正常，存活时间长达6 在无菌住房里呆了几个月。我们的初步数据显示，DKO小鼠 发展成一种更温和的自身免疫性结肠炎，可见于IL-2 单基因敲除小鼠。DKO小鼠可挂载强延迟型 过敏症。对外来蛋白质抗原的反应。最后，DKO 小鼠在B细胞成熟方面存在严重缺陷。我们的具体目标是 这一建议是为了(1)描述DKO小鼠的坐骑能力 细胞和体液免疫反应和(2)决定结果 无IL-2和IL-2的同种异体胰腺移植免疫反应 干扰素-γ(即排斥或耐受)。首先，我们将学习能力 对DKO小鼠产生迟发性超敏反应 组织相容性抗原(MHC)，在免疫后产生抗体， 并产生细胞毒性T淋巴细胞。第二，我们将呈现DKO小鼠 糖尿病患者使用链脲佐菌素，然后进行胰岛细胞 来自完全不同的MHC捐赠者的移植。同种异体移植的存活率将是 我们假设DKO小鼠会排斥同种异体胰岛移植， 排斥反应的机制与IL-4或其他T细胞有关 生长因子和细胞毒性T细胞。细胞因子环境与细胞 所涉及的表型将通过定量聚合酶链法进行鉴定 反应和荧光激活的细胞分选。相比之下，如果DKO 小鼠对同种异体胰岛移植有耐受性，我们将通过 耐受性T细胞亚群的过继转移。