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.

总体而言，我的实验室研究了同源趋化因子配体和趋化因子模拟物与G蛋白耦合趋化受体的相互作用，并激活受体，从而对炎症，免疫，自身免疫，自身免疫，癌症和Algesia产生影响。我们已经表明，多种抗菌肽（AMPS）模拟趋化因子，并且具有快速激活宿主免疫反应的能力。我们提议称这些预警信号警报器。警报的特征是表达GIPCR的细胞具有趋化活性，以及​​激活IDC以成熟为抗原呈递的能力，T淋巴细胞激活树突状细胞（MDC），从而产生了体内免疫辅助辅助效应。 警报蛋白的这些活性，如果与抗原一起给药，会导致细胞和体内对抗原的免疫反应的大量增强。我们先前以前将α和β类型的防御素鉴定为具有趋化性和激活对未成熟树突状细胞（IDC）（IDC）和体内免疫辅助效应的影响的警报蛋白。一些β防御素与CCR6趋化因子受体相互作用，而α防御素与尚未知道的G蛋白偶联受体（GIPCR）相互作用。另一种称为cathelicidin（LL37）及其鼠同源物绞痛的另一种抗菌肽是在IDC的单核细胞和前体表达的FPRL-1受体的趋化性，诱导IDC的成熟并诱导同样的辅助佐剂，在体内和铝中一样有效。尽管警报在结构上是不同的，但它们迅速从白细胞或受损细胞的颗粒中释放出来。响应于角质形成细胞的促炎刺激物或胃肠道，gu道和气管骨会树的上皮细胞也可以诱发警报蛋白。因此，警报蛋白可能代表了一种预警系统，该系统在上一年中提醒宿主防御机构危险信号，我们还调查了另一种白细胞颗粒衍生的警报器的免疫激活和趋化性效应，称为嗜酸性神经毒素（EDN），这是RNase家族的成员，是RNase家族的成员，并且具有抗病毒活性。 EDN基于其与百日咳毒素易感GIPCR的相互作用是IDC和MDC的趋化性。此外，基于与TLR2相互作用的EDN激活IDC，以产生多种促炎细胞因子并成熟成MDC。 EDN还具有有效的体内免疫刺激作用。但是，EDN以优先促进极化模式的方式激活了DC，而B-淋巴细胞更大地诱导了IgG1抗体产生，而生产白介素（IL）-5和13，但不是IL-4或IL-4或IFNGAMMA。尽管EDN具有警报的特性，但这表明EDN的佐剂作用可能证明在旨在针对寄生疾病而不是肿瘤的旨在免疫的疫苗中有用。我们还表明，乳铁蛋白具有警报活动。我们已经使用了人类乳铁蛋白（TLF）的GMP制剂进行研究。在其他中心，TLF正在评估为抗肿瘤治疗。 TLF是单核细胞APC的趋化性，但使用未鉴定的受体来募集和动员单核细胞。由于C3H/HEJ与C3H/HEN不同，因此TLF不会激活小鼠，因此这表明TLR4参与TLF的免疫辅助作用。 TLF活性似乎可以通过小鼠的胃肠道运输生存。因此，我们正在研究TLF是否可以加剧IBD，以及摄入乳铁蛋白是否会因此是否会诱发患有结肠癌的受试者。在过去的一年中，我们确定颗粒素通常存储在天然杀伤（NK）细胞和细胞毒性T细胞（CTL）的颗粒中，为警报。对于未成熟的髓样树突状细胞（IDC），颗粒素的9KDA和15KDA形式都是趋化的，并且可以将这种IDC激活至成熟到呈现抗原的树突状细胞（MDC）。 颗粒素与未鉴定的百日咳毒素敏感的GIα蛋白细胞受体（GIPCR）相互作用，以诱导趋化反应。像TLF一样，颗粒素也无法诱导C3H/HEJ IDC的成熟，这表明TLR4参与了颗粒素的免疫激活作用。内毒素污染可能解释了颗粒素的TLR4刺激作用的可能性已被排除在外。因此，我们可以将NK和CTL细胞添加到嗜酸性粒细胞，粒细胞，上皮细胞和巨噬细胞中，作为警报蛋白的来源。目前，我们正在制备融合蛋白，这些融合蛋白包括一些更有效的警报蛋白与几种不同的黑色素瘤肿瘤抗原融合，以获得有效的抗肿瘤疫苗。