COXSACKIEVIRUS AS VECTOR FOR ANTIINFLAMMATORY CYTOKINES

柯萨奇病毒作为抗炎细胞因子的载体

• 批准号: 2802433
• 负责人: Nora M. Chapman
• 金额: $ 14.03万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 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 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2802433
• 关键词: Coxsackievirus; antiviral antibody; attenuated microorganism; cytokine; heart disorder chemotherapy; helper T lymphocyte; idiopathic dilated cardiomyopathy; interleukin 10; interleukin 4; laboratory mouse; myocarditis; nonhuman therapy evaluation; polymerase chain reaction; tissue /cell culture; transfection /expression vector; virus replication

The coxsackie B virus cause acute myocarditis and dilated cardiomyopathy (DCM); DCM often results in a failing heart, the solution to which is generally transplantation to avoid eventual death. The long-term goal of this work is to determine whether genetically engineered cardiotropic CVB3 vectors expressing key cytokines will be useful as beneficial therapeutic, even prophylactic, approaches to diminishing inflammatory heart disease, including the ablation of rejection in heart transplantation. Our objective in this proposal is to evaluate and explore the efficacy of attenuated CVB3 strains, engineered to express the murine cytokines IL-4 of IL-10, in the diminution of symptoms of acute cardiac inflammatory disease. The specific aims are [1] Study attenuated CVB3 strains that express murine IL-4 (mIL-4) or mIL-10 in cell culture systems. Human and murine cell cultures will be used to evaluate the kinetics and extent of viral expression, genetic stability upon multiple passages, extent of export and biological activity of the expressed cytokines. [2] Study the murine immune response against the interleukin-expressing CVB3 strain. Mice will be evaluated for extent of viral replication in heart, extents and types of B and T cell responses, and how these compare temporally following virus inoculation. Cytokines specific for the Th1 and Th2 type T cell responses, in addition to mIL4 or mIL10, in murine sera and hearts following infection will be monitored and quantitated. RT-PCR analysis of cytokine gene expression in the murine host will also be analyzed as a function of the specific infecting virus. [3] Determine how CVB3-expressed mIL-4 or mIL-10 in mice affects progression and extent of CVB3-induced inflammatory heart disease. Extent of inflammation as a function of time will be photo microscopically mapped and quantitated. Extent and types of anti-viral T cell responses and circulating cytokine expression will be evaluated in test mice versus controls, as will types of anti- viral antibodies. Transient, low-level expression of key modulatory cyto- and chemokines by attenuated picornaviral vectors is a novel and potentially valuable new approach to treating serious inflammatory diseases.

科萨基B病毒引起急性心肌炎和扩张型心肌病 (DCM); DCM经常导致心脏衰竭，其解决方案是 一般是移植以避免最终死亡 远景目标 这项工作的目的是确定基因工程的心脏 表达关键细胞因子的CVB 3载体将是有用的， 治疗性甚至预防性的方法来减少炎症 心脏病，包括心脏排斥反应的消融 移植 我们在本提案中的目标是评估和 探索减毒CVB 3菌株的功效， 小鼠细胞因子IL-4或IL-10，在减轻 急性心脏炎性疾病。 具体目标是[1]研究 表达鼠IL-4（mIL-4）或mIL-10的减毒CVB 3株， 细胞培养系统 人和鼠细胞培养物将用于 评价病毒表达的动力学和程度、遗传稳定性 在多次传代后，输出的程度和生物活性 表达的细胞因子。 [2]研究小鼠针对 表达白细胞介素的CVB 3菌株。 将评价小鼠的程度 心脏中病毒复制的程度、B细胞和T细胞的类型 反应，以及这些如何比较病毒接种后的时间。 Th 1和Th 2型T细胞应答特异性细胞因子， 除了mIL 4或mIL 10，在感染后的鼠血清和心脏中 将被监测和量化。细胞因子基因的RT-PCR分析 还将分析鼠宿主中的表达作为免疫调节因子的函数。 特异性感染病毒 [3]确定CVB 3表达的mIL-4或 小鼠中的mIL-10影响CVB 3诱导的肝细胞凋亡的进展和程度 炎症性心脏病 炎症程度作为以下因素的函数 将对时间进行照片显微镜映射和定量。 程度和 抗病毒T细胞应答类型和循环细胞因子表达 将在试验小鼠与对照小鼠中进行评估，抗- 病毒抗体 关键调节因子的瞬时低水平表达 细胞因子和趋化因子的减毒小核糖核酸病毒载体是一种新的， 治疗严重炎症的潜在有价值的新方法 疾病