Characterization of Innate Binding Specificities of Peptidoglycan Binding Protein

肽聚糖结合蛋白的固有结合特异性的表征

• 批准号: 8882250
• 负责人: Andrew E. Levin
• 金额: $ 21.13万
• 依托单位: IMMUNETICS, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8882250
• 关键词: Antibodies; Applications Grants; Autoimmune Diseases; Bacillus cereus; Bacteria; Bacterial Infections; Basic Science; Binding; Binding Proteins; Binding Sites; Biological Assay; Blood Platelets; Buffers; Cell Wall; Cells; Chemistry; Cleaved cell; Clinical Research; Color; Complementary DNA; Densitometry; Detection; Diagnosis; Disease; Drosophila pros protein; Employee Strikes; Enterococcus faecalis; Enzymes; Escherichia coli; Excision; Goals; Gram-Negative Bacteria; Gram-Positive Bacteria; HIV; Health; Hemocytes; Hemolymph; High Pressure Liquid Chromatography; Human; Immune system; Inflammation; Insecta; Klebsiella pneumonia bacterium; Larva; Length; Lepidoptera; Lysostaphin; Malignant Neoplasms; Manduca; Manduca sexta; Measures; Membrane; Membrane Proteins; Meningitis; Methods; Microbe; Monophenol Monooxygenase; Muramic Acid; Muramidase; Mutate; Natural Immunity; Nosocomial Infections; Oklahoma; Oryctolagus cuniculus; Pathogen detection; Peptidoglycan; Performance; Polyacrylamide Gel Electrophoresis; Polysaccharides; Precipitation; Protein Binding; Proteins; Quality Control; Reaction; Recombinant Proteins; Recombinants; Research; Risk; Role; Sampling; Sepsis; Septic Shock; Serine Protease; Source; Specificity; Staphylococcus aureus; Supplementation; Surface; Technology; Testing; Thick; Transfusion; Universities; Vertebral column; base; blood product; complement system; crosslink; improved; innovation; microbial; mutanolysin; pathogen; peptidoglycan recognition protein; polyclonal antibody; pro-phenoloxidase; protein expression; research study; screening; systems research

DESCRIPTION (provided by applicant): BacTx(R) technology, developed by Immunetics for detection of bacteria, employs a unique and innovative assay chemistry to detect Gram-positive and Gram-negative bacteria. The technology is based on components of the innate immune system of the tobacco hornworm (Manduca sexta), in particular the prophenoloxidase cascade, in which serine proteases cleave prophenoloxidase to phenoloxidase, which subsequently oxidizes chromogen to generate a colored reaction product. The BacTx(R) Bacterial Detection Kit received 510(k) clearance from the FDA (CBER) in June 2012 as a quality control test for bacterial contamination in platelets. For M. sexta and other Lepidoptera, the most potent elicitor of the prophenoloxidase cascade is peptidoglycan, which is a major structural component of bacterial cell walls. In this application we propose to biochemically characterize the relationship between the presence of specific bacterial cell wall features and evasion of detection by pathogen recognition proteins present in M. sexta hemolymph. The role of these features in inhibiting prophenoloxidase activation will be investigated, with the long term goal of potentially enhancing assay performance and opening up new applications of BacTx(R) technology, including potential use for diagnosis of sepsis, meningitis, and nosocomial infections. Due to the striking similarities between the insect innate immune system and human complement system, this research may also prove significant to elucidating the relationship between inflammation due to bacterial infection and debilitating disorders such as cancer, autoimmune diseases, and septic shock.

描述（由申请人提供）：由免疫学用于检测细菌的BACTX（R）技术采用独特而创新的测定化学方法来检测革兰氏阴性和革兰氏阴性细菌。该技术基于烟草虫（Manduca sexta）的先天免疫系统的组成部分，尤其是预防性氧化酶级联反应，其中丝氨酸蛋白酶将预防性氧化酶裂解至苯酚氧化酶，随后将其氧化为氧化以产生彩色反应产物。 BACTX（R）细菌检测试剂盒在2012年6月从FDA（CBER）获得了510（k）的清除，作为血小板细菌污染的质量控制测试。对于M. sexta和其他鳞翅目而言，预测酶级联反应的最有效的引起者是肽聚糖，它是细菌细胞壁的主要结构成分。在此应用中，我们建议以生化为特征 在特定细菌细胞壁特征的存在与sexta Hylymph中存在的病原体识别蛋白逃避检测之间。这些特征在抑制预防氧化酶激活中的作用将被研究，其长期目标是潜在的 提高测定性能并开放BACTX（R）技术的新应用，包括潜在用来诊断败血症，脑膜炎和医院感染。由于昆虫先天免疫系统与人类补体系统之间的惊人相似之处，这项研究也可能对阐明由于细菌感染和使人衰弱的疾病（如癌症，自身免疫性疾病和败血性休克）引起的炎症之间的关系显着。