Mechanism of Action of Inhibitory CpG Oligonucleotides

抑制性 CpG 寡核苷酸的作用机制

• 批准号: 7407449
• 负责人: PETAR S LENERT
• 金额: $ 35.13万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7407449
• 关键词: Address; Affinity; Apoptosis; Appendix; Autoimmune Diseases; B-Lymphocytes; Bacteria; Bacterial DNA; Binding; Biology; Cell Cycle; Cells; Colitis; DNA; Dendritic Cells; Development; Dimerization; Disease; Family; Grant; Human; Hypersensitivity; Immune; Immune response; Immune system; Immunity; In Vitro; Inflammatory; Libraries; Lupus; Malignant Neoplasms; Microbe; Modeling; Modification; Molecular; Mus; Oligodeoxyribonucleotides; Oligonucleotides; Pathology; Pattern; Play; Positioning Attribute; Range; Recombinants; Rheumatoid Arthritis; Role; Septic Shock; Series; Shock; Signal Transduction; Specificity; Structure-Activity Relationship; TLR9 gene; Therapeutic; Toll-like receptors; Transcription Factor AP-1; Translating; Vaccine Adjuvant; Viral Tumor Antigens; base; cell type; cytokine; design; human disease; in vivo; inhibitor/antagonist; macrophage; microbial; prevent; research study; response

The CpG motifs of bacterial DMA are among the microbial molecular patterns recognized as "danger signals" by our innate immune system, requiring Toll-Like Receptor (TLR) 9. Short synthetic oligodeoxyribonucleotides (ODN) have facilitated studies of structure-function relationships. The ability of stimulatory (ST-) ODN to activate B cells, macrophages, and dendritic cells directly, thus triggering humoral and TH1 immunity together, gives them valuable potential roles as vaccine adjuvants and as therapy in cancer and allergy. Changing as few as 2 bases converts a strong ST-ODN into an inhibitor (IN-ODN) that blocks TLR9 but not the other TLRs. With support from the current grant the sequence requirements for IN- ODN action in mouse cells have been defined. Only 3 pairs of positions out of 15 determine IN-ODN activity. The same panel of IN-ODN can block the ST-ODN preferred by B cells (Type B) and non-B cells (Type A). We request continued support in order to address these questions: 1) Do IN-ODN compete with ST-ODN for TLR9 binding? 2) Does the strength of binding of ST-ODN and IN-ODN for TLR9 determine their biologic activity? 3) Do ST-ODN or IN-ODN tie TLR9 molecules together? Does the ability of ODN to aggregate with themselves or each other allow them to do this? 4) Where in the cell does the competition between ST-ODN and IN-ODN take place? 5) Can IN-ODN block ST-ODN activity in vivo? Do the same subtle structural requirements apply? Examples to be used are a model of ODN-induced "septic shock" and an inflammatory colitis in mice. 6) What are the IN-ODN structural requirements for human cells? Do they differ between ODN types? The answers to these questions are pertinent both to understanding the mechanism of ODN responses and to developing further applications to human disease, especially the prospect of treating lupus and rheumatoid arthritis with IN-ODN.

细菌DNA的CpG基序是被认为是“危险信号”的微生物分子模式之一 通过我们的先天免疫系统，需要Toll样受体（TLR）9。短合成 寡脱氧核糖核苷酸（ODN）促进了结构-功能关系的研究。的能力 刺激性（ST-）ODN直接激活B细胞、巨噬细胞和树突状细胞，从而触发体液免疫。 和TH1免疫结合在一起，使它们作为疫苗佐剂和治疗 癌症和过敏改变少至2个碱基将强ST-ODN转化为抑制剂（IN-ODN）， 阻断TLR9而不阻断其它TLR。在目前赠款的支持下，IN- ODN在小鼠细胞中的作用已被定义。15个位点中只有3对决定IN-ODN活性。 同一组IN-ODN可以阻断B细胞（B型）和非B细胞（A型）优选的ST-ODN。 我们要求继续提供支持，以解决这些问题： 1)IN-ODN与ST-ODN竞争TLR9结合吗？ 2)ST-ODN和IN-ODN与TLR9的结合强度是否决定了它们的生物活性？ 3)ST-ODN或IN-ODN是否将TLR9分子连接在一起？ODN聚合与 他们自己或彼此允许他们这样做吗？ 4)ST-ODN和IN-ODN之间的竞争发生在小区的什么地方？ 5)IN-ODN能在体内阻断ST-ODN活性吗？同样微妙的结构要求是否适用？ 使用的实例是ODN诱导的"败血性休克"模型和小鼠中的炎性结肠炎。 6)人类细胞的IN-ODN结构要求是什么？ODN类型之间有区别吗？ 这些问题的答案对于理解ODN反应的机制和 并涉及开发对人类疾病的进一步应用，特别是治疗狼疮的前景， 类风湿性关节炎与IN-ODN。