FOCUSING IMMUNITY vs BOTULINUM TOXIN WITH CYTOKINE DNA

使用细胞因子 DNA 聚焦免疫与肉毒杆菌毒素

• 批准号: 6862685
• 负责人: DAVID S STRAYER
• 金额: $ 50.34万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6862685
• 关键词: Adenoviridae; Clostridium botulinum; active immunization; antigen antibody reaction; biotechnology; bioterrorism /chemical warfare; botulinum toxins; complementary DNA; cooperative study; cytokine; gene delivery system; immunoglobulin A; interleukin 4; interleukin 5; laboratory mouse; mucosal immunity; plasmids; transfection /expression vector; transforming growth factors; vaccine development; vector vaccine

DESCRIPTION (provided by applicant): The toxin of Clostridium botulinum, particularly administered by inhalation, is a potential weapon of bioterrorism and germ warfare, and protecting people from the potentially devastating effects of its use is a high priority. At the same time, botulinum toxin (BT) has very important medicinal uses as an antispasmotic agent. We propose a collaborative project to use gene delivery of cDNAs encoding BT, serotype A (BT/A), antigens, to be given in tandem with cDNAs encoding cytokines that preferentially stimulate secretory IgA antibody (SIgA) responses. The goal of these studies is develop immunization regimens that protect from inhalational BT without precluding the important therapeutic uses of this agent. We will use both plasmid and recombinant adenoviral gene delivery vehicles to deliver both the heavy chain of BT (HC). Antibody elicited by HC protects from challenge with the complete toxin. These vectors will also deliver DNAs encoding cytokines that preferentially weight immune responses in favor of SIgA, at the expense of serum IgG antibody: transforming growth factor-beta1 and interleukins-4 and -5 (IL-4, IL-5). Our main hypothesis is: Immunizing regimens using cytokines that favor SIgA can elicit protective mucosal immunity to BT without system antibody. To test this hypothesis we propose four aims. We will: 1. Produce plasmid and adenovirus expression constructs to deliver BT/A-HC and cvtokine cDNAs. 2. Define parameters of local and svstemic immunity to BT, elicited by plasmid and rAd vectors. 3. Add cytokine-carrying vectors, individually and then in combinations, to these immunization regimens. 4. Modify expression constructs and administration regimens to optimize selectivity, protection, and safety. Thus, we will apply current understanding of the signaling mechanisms that occur during the activation of the secretory immune system to the need to protect people from the potential weaponization of botulinum toxin without limiting its important therapeutic applications.

描述（由申请人提供）：肉毒杆菌毒素，尤其是通过吸入施用的毒素，是生物恐怖主义和细菌战的潜在武器，保护人们免受其使用造成的潜在破坏性影响是当务之急。同时，肉毒杆菌毒素（BT）作为解痉剂具有非常重要的药用用途。我们提出了一个合作项目，利用编码 BT、A 血清型 (BT/A) 抗原的 cDNA 进行基因递送，与编码优先刺激分泌型 IgA 抗体 (SIgA) 反应的细胞因子的 cDNA 一起给予。这些研究的目标是开发免疫方案，在不妨碍该药物重要治疗用途的情况下预防吸入性 BT。我们将使用质粒和重组腺病毒基因递送载体来递送 BT (HC) 的重链。 HC 引发的抗体可防止完整毒素的攻击。这些载体还将递送编码细胞因子的 DNA，这些细胞因子优先权衡有利于 SIgA 的免疫反应，但会牺牲血清 IgG 抗体：转化生长因子-β1 以及白细胞介素-4 和 -5（IL-4、IL-5）。我们的主要假设是： 使用有利于 SIgA 的细胞因子的免疫方案可以在没有系统抗体的情况下引发针对 BT 的保护性粘膜免疫。 为了检验这一假设，我们提出了四个目标。我们将： 1. 产生质粒和腺病毒表达构建体以递送 BT/A-HC 和细胞因子 cDNA。 2. 定义由质粒和 rAd 载体引发的 BT 局部和全身免疫参数。 3. 将携带细胞因子的载体单独添加，然后组合添加到这些免疫方案中。 4. 修改表达构建体和给药方案以优化选择性、保护性和安全性。 因此，我们将应用目前对分泌性免疫系统激活过程中发生的信号机制的理解来满足保护人们免受肉毒杆菌毒素潜在武器化的需要，而不限制其重要的治疗应用。