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Development of immune supression against gene product in gene therapy

基因治疗中针对基因产物的免疫抑制的发展

基本信息

批准号：
11557066
负责人：
AMAGAI Masayuki
金额：
$ 8.51万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557066/
关键词：
Gene therapy Inherited disease Immune response Autoimmune Model mouse Knockout mouse Skin graft CD40 ligand 遺伝性皮膚疾患免疫寛容経口免疫経鼻免疫

项目摘要

Gene therapy to treat recessive genodermatoses may provoke unwanted immune response against the introduced gene product because tolerance against this novel protein is absent in patients. In this study, using desmoglein 3 knockout (Dsg3-/-) mice as a disease model for genetic defect of DSG3, we investigated whether immune response against Dsg3 is provoked by nonviral gene therapy and whether such reaction could be prevented. When mouse Dsg3 CDNA was introduced in the skin of Dsg3-/- mice by naked DNA injection, some Dsg3-/- mice developed anti-Dsg3 IgG which bound to Dsg3 expressed by the therapy in vivo. To overcome this problem, we used anti-CD40L monoclonal antibody (MR1) to block co-stimulatory interaction between CD40-CD40L, which is important in the triggering and maintenance of immune responses. For this assay, we employed the Dsg3+/+ skin graft on Dsg3-/- mice to represent stable gene transfer of Dsg3. After skin grafting, all recipient Dsg3-/- mice were treated either with MR1 or with hamster IgG regularly. All of hamster IgG treated mice developed circulating anti-Dsg3 IgG 2-3 weeks after the skin graft. This IgG production lasted: more than 4 weeks and IgG deposition was observed on the surfaces of the keratinocytes in grafts.; However, this anti-Dsg3 IgG production was efficiently suppressed when the recipient mice were treated with MR1. These findings indicate that effective prevention of such undesirable immune response is required for a successful gene therapy for recessive genodermatoses and that the blockade of CD40-CD40L interaction may be a valuable way to achieve this prevention.

治疗隐性遗传性皮肤病的基因疗法可能会引起对引入的基因产物的不必要的免疫反应，因为患者对这种新蛋白缺乏耐受性。本研究以桥粒芯糖蛋白3基因敲除小鼠(Dsg3-/-)作为DSG3基因缺陷的疾病模型，探讨非病毒基因治疗是否能引起对Dsg3的免疫应答，以及这种反应是否可以被预防。将裸鼠Dsg3 CDNA导入Dsg3-/-小鼠皮肤后，部分Dsg3-/-小鼠体内产生抗Dsg3抗体，并与治疗后表达的Dsg3结合。为了克服这一问题，我们使用抗CD40L单抗(MR1)来阻断CD40-CD40L之间的共刺激相互作用，这在免疫应答的触发和维持中是重要的。在这项实验中，我们使用Dsg3+/+皮肤移植物在Dsg3-/-小鼠身上代表Dsg3稳定的基因转移。皮肤移植后，所有受体Dsg3-/-小鼠均定期接受MR1或仓鼠免疫球蛋白治疗。所有经金黄地鼠免疫球蛋白处理的小鼠在皮肤移植后2-3周均出现循环抗Dsg3抗体。这种免疫球蛋白的产生持续了4周以上，在移植物的角质形成细胞表面观察到了免疫球蛋白的沉积。然而，当接受MR1治疗时，这种抗Dsg3的免疫球蛋白的产生被有效地抑制。这些发现表明，有效地预防这种不良的免疫反应是成功的隐性遗传性皮肤病基因治疗所必需的，而阻断CD40-CD40L相互作用可能是实现这一预防的有价值的方法。