Granulysin Derived Immunotherapeutics for Biodefense

说明(申请人提供)：防御与在恐怖主义或战争行为中使用生物制剂有关的感染是国家优先事项。为了实现这一目标，必须开发新的方法来检测、治疗和预防感染A-C类病原体。治疗可包括在面临直接威胁的情况下预防感染，保护免疫受损的个人，或暴露后治疗以抑制感染和疾病。用于这些应用的理想试剂应该是稳定、广谱、快速、对人体细胞无毒、廉价和易于制造的试剂。传统抗生素符合这些标准中的一部分，但耐药生物体的增加和新的广泛使用的抗生素的缺乏都使识别新的抗菌素成为当务之急。颗粒溶素是一种由人类自然杀伤细胞和活化的T淋巴细胞表达的α螺旋蛋白。重组颗粒溶素既能裂解哺乳动物细胞，又能裂解多种微生物。与颗粒溶素中心区域相对应的合成肽(10-30个残基)概括了其裂解活性。在这些多肽的一个子集中，半胱氨酸或精氨酸残基的替换，或者引入D-氨基酸来破坏α-螺旋，会导致对哺乳动物细胞的活性丧失，而对抗菌活性几乎没有影响。该计划的目标是开发基于这些颗粒溶素肽的新型免疫疗法。其他的衍生物将在项目1中产生，在项目2中进行体外和体内评估，并在项目3中描述作用机制。这些项目将得到三个核心的支持--合成/库存；BSL-3设施和管理。从这些研究中收集到的信息应该有助于开发新的免疫疗法，用于生物防御和治疗抗生素耐药病原体。

DESCRIPTION (provided by applicant): Defense against infections related to the use of biological agents in acts of terrorism or war is a national priority. To achieve this end, new methods to detect, treat, and prevent infection with Category A-C pathogens must be developed. Treatment may include prevention of infection in the case of an immediate threat, protection of immunocompromised individuals, or post-exposure treatment to suppress infection and disease. Ideal agents for these applications would be stable, broad spectrum, fast acting, nontoxic towards human cells, inexpensive, and easy to manufacture. Conventional antibiotics meet some of these criteria, but both the rise in antibiotic resistant organisms and a dearth of new broad-based antibiotics make identification of new antimicrobials imperative. Granulysin is an alpha-helical protein expressed by human natural killer cells and activated T lymphocytes. Recombinant granulysin lyses both mammalian cells and a broad spectrum of microbes. Synthetic peptides (10-30 residues) corresponding to the central region of granulysin recapitulate its lytic activity. In a subset of these peptides, replacement of cysteine or arginine residues, or introduction of D-amino acids to disrupt the alpha-helix, results in the loss of activity against mammalian cells with little or no effect on antimicrobial activity. The goal of this Program is to develop novel immunotherapeutics based on these granulysin peptides. Additional derivatives will be generated in Project 1, evaluated in vitro and in vivo in Project 2, and characterized for mechanism of action in Project 3. These projects will be supported by three cores--synthesis/inventory; a BSL-3 facility, and administration. The information gleaned from these studies should lead to the development of new immunotherapeutics for biodefense and for treatment of antibiotic resistant pathogens.