FOCUSING IMMUNITY vs BOTULINUM TOXIN WITH CYTOKINE DNA

描述(申请人提供)：肉毒梭菌毒素，特别是通过吸入给予，是潜在的生物恐怖主义和细菌战的武器，保护人们免受其使用的潜在破坏性影响是高度优先的。同时，肉毒杆菌毒素(Bt)作为一种重要的解痉剂具有重要的药用价值。我们提出了一个合作项目，利用编码Bt血清A型(Bt/A)抗原的cDNA与编码优先刺激分泌性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，在不排除这种制剂的重要治疗用途的情况下防止吸入Bt。我们将使用质粒和重组腺病毒基因载体来传递Bt(HC)的重链。由HC诱导的抗体可保护机体免受完全毒素的攻击。这些载体还将传递编码细胞因子的DNA，这些细胞因子优先加重免疫应答，有利于SIgA，以牺牲血清免疫球蛋白抗体：转化生长因子-β1和白介素4和-5(IL-4，IL-5)为代价。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诱导对Bt的保护性黏膜免疫。 为了检验这一假设，我们提出了四个目标。我们会： 1.构建携带Bt/A-HC和cvtokine基因的表达载体和腺病毒表达载体。 2.确定Bt的局部和血吸性免疫参数，分别由质粒和Rad载体诱导。 3.将细胞因子载体单独和组合添加到这些免疫方案中。 4.修改表达结构和给药方案，以优化选择性、保护和安全性。 因此，我们将把目前对分泌免疫系统激活过程中发生的信号机制的理解应用于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。

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.