FOCUSING IMMUNITY vs BOTULINUM TOXIN WITH CYTOKINE DNA

描述（由申请人提供）：肉毒梭菌毒素，特别是通过吸入给药，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的潜在破坏性影响是一个高度优先事项。同时，肉毒杆菌毒素（BT）作为一种解痉剂具有非常重要的医学用途。我们提出了一个合作项目，使用基因传递的cDNA编码BT，血清型A（BT/A），抗原，给予串联的cDNA编码细胞因子，优先刺激分泌型伊加抗体（SIgA）的反应。这些研究的目的是开发免疫方案，保护免受吸入性BT，而不排除这种药物的重要治疗用途。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。由HC引发的抗体保护免受完整毒素的攻击。这些载体还将递送编码细胞因子的DNA，所述细胞因子以牺牲血清IgG抗体为代价优先加重有利于SIgA的免疫应答：转化生长因子-β 1和白细胞介素-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起对BT的保护性粘膜免疫。 为了验证这一假设，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cvtokine cDNA。 2.定义由质粒和rAd载体引起的对BT的局部和全身免疫的参数。 3.在这些免疫方案中，单独地然后组合地添加携带麻黄碱的载体。 4.修改表达构建体和给药方案以优化选择性、保护和安全性。 因此，我们将应用当前对分泌免疫系统激活过程中发生的信号传导机制的理解，以保护人们免受肉毒杆菌毒素潜在武器化的影响，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.