Studies of Chemokine-Receptor Interactions with Chemokines and alarmins

趋化因子受体与趋化因子和警报素相互作用的研究

• 批准号: 7965166
• 负责人: JOOST J OPPENHEIM
• 金额: $ 115.53万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7965166
• 关键词: Accounting; Adjuvant; Antibody Formation; Antigens; Antiviral Agents; Autoimmunity; B-Lymphocytes; C3H/HeN Mouse; CCR6 gene; Cell Line; Cells; Chimeric Proteins; Colon Carcinoma; Coupled; Cytoplasmic Granules; Cytotoxic T-Lymphocytes; Defensins; Dendritic Cells; Development; Endotoxins; Eosinophil-Derived Neurotoxin; Epithelial Cells; Family; G-Protein-Coupled Receptors; GTP-Binding Proteins; Gastrointestinal tract structure; Gi-alpha protein; Homologous Gene; Host Defense; Human; IgG1; Immune; Immune response; Immunity; Immunization; Immunologic Adjuvants; Infection; Inflammation; Ingestion; Injury; Interleukin-4; Interleukin-5; Laboratories; Lactoferrin; Leukocytes; Ligands; Malignant Neoplasms; Mononuclear; Mus; Myelogenous; Natural Killer Cells; Organism; Pain; Parasitic Diseases; Pattern; Pertussis Toxin; Preparation; Production; Property; Receptor Cell; Recruitment Activity; Signal Transduction; Source; System; T-Lymphocyte; TLR2 gene; TLR4 gene; Therapeutic; Tissues; Tracheobronchial; Trauma; Trees; Tumor Antigens; Vaccines; alpha-Defensins; aluminum sulfate; antimicrobial peptide; base; beta-Defensins; cathelicidin; cell injury; chemokine; chemokine receptor; cytokine; eosinophil; granulocyte; granulysin; in vivo; keratinocyte; macrophage; melanoma; member; microbial; monocyte; receptor; response; tumor; tumor growth

Overall, my laboratory investigates the interactions of cognate chemokine ligands and chemokine mimics with G-protein coupled chemotactic receptors and activating receptors with resultant effects on inflammation, immunity, autoimmunity, cancer and algesia. We have shown that a variety of antimicrobial peptides (AMPs) mimic chemokines and also have the capacity to rapidly activate host immune responses. We have proposed calling these early warning signals alarmins. Alarmins are characterized by having chemotactic activity for cells expressing GiPCR, together with the capacity to activate iDC to mature into antigen- presenting, T lymphocyte activating dendritic cells (mDC) with resultant in vivo immunoadjuvant effects. These activities of alarmins, if administered together with an antigen, result in considerable augmentation of both cellular and humoral in vivo immune responses to the antigen. We previously identified both alpha and beta types of defensins as alarmins with chemotactic and activating effects on immature dendritic cells (iDC) and in vivo immunoadjuvant effects. Some of the beta defensins interact with the CCR6 chemokine receptor, while alpha defensins interact with an as yet unknown G-Protein Coupled Receptors (GiPCR). Another antimicrobial peptide known as cathelicidin (LL37) and its murine homologue CRAMP are chemotactic for FPRL-1 receptors expressed on monocytes and precursors of iDC, induce the maturation of iDC and are equally as potent adjuvants in vivo as alum. Although alarmins are structurally distinct, they are rapidly released from granules of leukocytes or damaged cells. Alarmins can also be induced in response to proinflammatory stimulants by keratinocytes or epithelial cells lining the GI tract, GU tract and tracheobronchial tree. As such, alarmins probably represent an early warning system to alert the host defense to danger signals During the previous year we also investigated the immune activating and chemotactic effects of another leukocyte granule derived alarmin known as eosinophil derived neurotoxin (EDN), which is a member of the RNAse family, and has antiviral activity including anti-HIV activity. EDN based on its interactions with a pertussis toxin susceptible GiPCR is chemotactic for iDC, and mDC. In addition, EDN based on interactions with TLR2 activates iDC to produce multiple proinflammatory cytokines and to mature into mDC. EDN also has potent in vivo immunostimulating effects. However, EDN activated DC in a manner that preferentially promotes a Th2 pattern of polarization with greater induction of IgG1 antibody production by B-lymphocytes and the production of Interleukin (IL)-5 and 13, but not IL-4 or IFNgamma. Although EDN has the properties of an alarmin this suggests that the adjuvant effect of EDN may prove more useful in vaccines aimed at immunization against parasitic diseases rather than tumors. We have also shown that lactoferrin has alarmin activities. We have used GMP preparations of human lactoferrin (TLF) for our studies. TLF is being evaluated as an anti-tumor therapeutic at other centers. TLF is chemotactic for monocytic APCs, but uses an unidentified receptor to recruit and mobilize mononuclear cells. Since C3H/HeJ, unlike C3H/HeN, mice are not activated by TLF, this suggests TLR4 is involved in the immune adjuvant effects of TLF. TLF activity appears to survive transport through the gastrointestinal tract of mice. We are therefore investigating whether TLF can exacerbate IBD and whether ingestion of lactoferrin may thus predispose subjects to develop colon cancer. During the past year we identified granulysin, which is normally stored in the granules of natural killer (NK) cell and cytotoxic T cells (CTL), as an alarmin. Both 9kDa and 15kDa forms of granulysin are chemotactic for immature myeloid derived dendritic cells (iDC) and can activate such iDC to mature into antigen presenting dendritic cells (mDC). Granulysin interacts with an unidentified pertussis toxin sensitive Gi alpha protein cell receptor (GiPCR) to induce chemotactic responses. Like TLF, granulysin also fails to induce maturation of C3H/HeJ iDC, suggesting TLR4 is involved in the immune activating effect of granulysin. The possibility that endotoxin contamination accounts for the TLR4 stimulating effect of granulysin has been ruled out. Consequently, we can add NK and CTL cells to eosinophils, granulocytes, epithelial cells and macrophages as sources of alarmins. We are at present preparing fusion proteins consisting of some of the more potent alarmins fused to several different melanoma tumor antigens in order to obtain a potent antitumor vaccine.

总的来说，我的实验室研究了同源趋化因子配体和 具有G蛋白偶联的趋化性受体和活化受体的趋化因子模拟物， 对炎症、免疫、自身免疫、癌症和痛觉产生的影响。我们已经表明 各种抗菌肽（AMP）模拟趋化因子，并且也具有 迅速激活宿主免疫反应。我们建议称这些早期预警信号为 危言耸听Alarmin的特征在于对表达以下蛋白的细胞具有趋化活性： GiPCR，连同激活iDC成熟为抗原呈递的能力，T 淋巴细胞活化树突状细胞（mDC），产生体内免疫佐剂作用。 如果与抗原一起施用，则警报素的这些活性导致相当大的免疫应答。 增强对抗原的细胞和体液体内免疫应答。我们 先前鉴定出α和β型防御素都是具有趋化性的警报素， 对未成熟树突细胞（iDC）的活化作用和体内免疫佐剂作用。一些 的β防御素与CCR 6趋化因子受体相互作用， G蛋白偶联受体（GiPCR）。另一抗微生物 称为凯萨林菌素（LL 37）的肽及其鼠同源物CRAMP对FPRL-1具有趋化性 单核细胞和iDC前体上表达的受体诱导iDC成熟， 在体内与明矾一样作为有效的佐剂。尽管警报素在结构上不同，但它们 从白细胞或受损细胞的颗粒中迅速释放出来。警报器也可以是 由角质形成细胞或上皮细胞衬里的促炎刺激物诱导 胃肠道、胃肠道和气管支气管树。因此，警报器可能代表一个 预警系统，以提醒主机防御危险信号在过去的一年里，我们 还研究了另一种白细胞颗粒的免疫激活和趋化作用 嗜酸性粒细胞衍生的神经毒素（EDN），其是RNA酶的成员。 家族，并且具有抗病毒活性，包括抗HIV活性。基于其相互作用的EDN 对百日咳毒素敏感的GiPCR对iDC和mDC具有趋化性。此外，EDN 基于与TLR 2的相互作用激活iDC产生多种促炎细胞因子 并成熟为mDC。EDN还具有有效的体内免疫刺激作用。然而，EDN 以优先促进Th 2极化模式的方式激活DC， B淋巴细胞产生的IgG 1抗体的诱导作用更强， 白细胞介素（IL）-5和13，但不是IL-4或IFN γ。虽然EDN具有 这表明EDN的佐剂作用可能在针对性的疫苗中更有用。 寄生虫病而不是肿瘤的免疫。我们还表明， 乳铁蛋白具有报警活性。我们已经使用人乳铁蛋白（TLF）的GMP制剂 为了我们的研究。TLF正在其他中心作为抗肿瘤治疗进行评估。TLF是 单核细胞APC的趋化性，但使用未鉴定的受体来招募和动员 单核细胞由于C3 H/HeJ与C3 H/HeN不同，小鼠不被TLF激活，这表明 TLR 4参与TLF的免疫佐剂作用。TLF活动似乎在 通过小鼠胃肠道的运输。因此，我们正在调查是否 TLF可加重IBD，以及摄入乳铁蛋白是否因此使受试者易患IBD。 患上结肠癌在过去的一年里，我们发现了颗粒溶素，它通常储存在 在自然杀伤（NK）细胞和细胞毒性T细胞（CTL）的颗粒中，作为警报素。两 9 kDa和15 kDa形式的颗粒溶解素对未成熟髓源性树突状细胞具有趋化性 树突状细胞（mDC）是树突状细胞（iDC）中的一种，并且可以激活这样的iDC以成熟为抗原呈递树突状细胞（mDC）。 颗粒溶素与未鉴定的百日咳毒素敏感性Gi α蛋白细胞相互作用 受体（GiPCR）来诱导趋化反应。与TLF一样，颗粒溶解素也不能诱导 C3 H/HeJ iDC的成熟，表明TLR 4参与C3 H/HeJ iDC的免疫激活作用。 颗粒溶素。内毒素污染导致TLR 4刺激的可能性 排除了颗粒溶解素的影响。因此，我们可以添加NK和CTL细胞， 嗜酸性粒细胞、粒细胞、上皮细胞和巨噬细胞作为警报素的来源。我们正处于 本发明提供了制备融合蛋白的方法，所述融合蛋白由一些更有效的alarmin融合到 几种不同的黑色素瘤肿瘤抗原，以获得有效的抗肿瘤疫苗。