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.

Coxsackie B病毒引起急性心肌炎和扩张的心肌病 （DCM）; DCM通常会导致心脏失败，而解决方案是 通常移植以避免最终死亡。 长期目标 这项工作是确定基因工程性心脏的 表达关键细胞因子的CVB3载体将作为有益 治疗性，甚至预防性，可减少炎症 心脏病，包括心脏拒绝的消融 移植。 我们在此提案中的目标是评估和 探索衰减的CVB3菌株的功效，设计为表达 IL-10的鼠细胞因子IL-4，症状减轻 急性心脏炎症性疾病。 具体目的是[1]研究 表达鼠IL-4（MIL-4）或MIL-10的衰减CVB3菌株 细胞培养系统。 人类和鼠类细胞培养物将用于 评估病毒表达的动力学和程度，遗传稳定性 在多个段落中，出口和生物活性的范围 表达的细胞因子。 [2]研究鼠免疫反应针对 表达白介素的CVB3菌株。 小鼠将在程度上评估 B和T细胞的心脏，范围和类型的病毒复制 反应，以及这些病毒接种后如何比较时间。 针对Th1和Th2型T细胞反应的细胞因子，在 在感染后，在MIL4或MIL10中增加了鼠血清和心脏 将受到监视和定量。 RT-PCR细胞因子基因分析 在鼠宿主中的表达也将作为与 特异性感染病毒。 [3]确定CVB3表达的MIL-4或 小鼠中的MIL-10影响CVB3诱导的进展和程度 炎症性心脏病。 炎症程度作为 时间将是微观映射和定量的照片。 范围和 抗病毒T细胞反应和循环细胞因子表达的类型 将在测试小鼠与对照组中评估，抗抗 病毒抗体。 关键调节的瞬态，低级表达 衰减的picornaviral载体的细胞和趋化因子是一种新颖， 潜在的有价值的新方法来治疗严重的炎症 疾病。