Consequences of Chemokine-Receptor Interactions with Chemokines and Chemokine Mi

趋化因子受体与趋化因子和趋化因子 Mi 相互作用的后果

• 批准号: 7592610
• 负责人: JOOST J OPPENHEIM
• 金额: $ 77.99万
• 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/7592610
• 关键词: Adjuvant; Amino Acyl-tRNA Synthetases; Antibodies; Antibody Formation; Antigens; Antiviral Agents; Asparagine-tRNA ligase; Autoantigens; Autoimmune Process; Autoimmunity; B-Lymphocytes; BLR1 gene; Boxing; Brugia malayi; CC chemokine receptor 3; CCR5 gene; CCR6 gene; CTAG1 gene; CXCR3 gene; Cancer Vaccines; Cell Line; Cell membrane; Cells; Cholesterol; Coupled; Cytoplasmic Granules; DNA-Binding Proteins; Defensins; Dendritic Cells; Development; Disruption; Embryo; Eosinophil-Derived Neurotoxin; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Epithelial Cells; Exhibits; Experimental Autoimmune Encephalomyelitis; Family; G-Protein-Coupled Receptors; GRB10 gene; GTP-Binding Proteins; Gastrointestinal tract structure; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; HMGB1 gene; Histidine-tRNA Ligase; Homologous Gene; Host Defense; Human; IL8RA gene; IL8RB gene; IgG1; Immune; Immune response; Immunization; Immunologic Adjuvants; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Injury; Insulin-Dependent Diabetes Mellitus; Interferon Type II; Interleukin-12; Interleukin-4; Interleukin-5; Laboratories; Lead; Leukocytes; Ligands; Malignant Neoplasms; Mammary Neoplasms; Membrane Microdomains; Mucin-1 Staining Method; Mus; Myositis; Neoplasm Metastasis; Nuclear; Numbers; Organism; Pain; Parasites; Parasitic Diseases; Patients; Pattern; Pertussis Toxin; Production; Property; Proteins; Reporting; Scleroderma; Signal Transduction; Site; System; T-Lymphocyte; TLR2 gene; Testing; Tissues; To autoantigen; Tracheobronchial; Trauma; Trees; Tumor Antigens; Vaccines; alpha-Defensins; aluminum sulfate; antimicrobial peptide; autoimmune uveitis; base; beta-Defensins; cancer cell; cathelicidin; chemokine; chemokine receptor; cytokine; immunogenic; in vivo; keratinocyte; member; methyl-beta-cyclodextrin; microbial; monocyte; neoplastic cell; receptor; response; trafficking; tumor; tumor growth

Overall, my laboratory investigates the interactions of cognate chemokine ligands and chemokine mimics with G-protein coupled chemokine receptors and the resultant effects on inflammation, autoimmunity, cancer and algesia. We have shown that a variety of antimicrobial peptides 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 leukocytes expressing GiPCR, together with the capacity to induce 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 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. 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 secreted in response to proinflammatory stimulants by keratinocytes or epithelial cells lining the GI tract, GU tract and tracheobronchial tree. As such, alarmins probably represent the early warning system to alert the host defense to danger signals. Like GM-CSF, a cytokine which like alarmins has chemotactic effect on dendritic cells and immunoadjuvant effects, these alarmins may prove useful as adjuvants in tumor vaccines. During the current year we have 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 to exhibit 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 High Mobility Group Box I (HMGB1), a nuclear (DNA) binding protein with antiviral activity, is chemotactic for a GiPCR on iDC. Our collaborators, Dr. Kevin Tracey, et al, have shown HMGB1 to be a potent inducer of proinflammatory cytokines, to induce the maturation of iDC to mDC, and to be highly produced at inflammatory sites and to have immunoadjuvant effects. Thus, HMGB1 is a potent alarmin, and since it induces the in vitro production of cytokines such as IL-12 which in turn induces IFN gamma, HMGB1 as an inducer of TH1 immune responses may prove to be a potent adjuvant for antitumor vaccine. We will test whether some of the alarmins we have identified can convert tolerogenic to immunogenic responses, which would be desirable in the case of tolerizing tumors. GiPCR not only interact with non-cognate alarmins, but also are responsible for the chemotactic responses of iDC to many autoantigens and some tumor antigens. We previously reported that histidyl tRNA synthetase (HRS) and AsnRS, autoantigens to which some patients with myositis develop auto-antibodies, are chemotactic for cells (including iDC) expressing CCR5 and CCR3 receptors respectively. More recently we have shown that of the antigens that induce experimental autoimmune uveitis (EAU), IRBP uses CXCR3 and CXCR5, while S-antigen uses only CXCR3 to chemoattract iDC. Many other autoantigens associated with type I diabetes, EAE and scleroderma are also chemotactic for iDC. Unrelated tRNA synthetases and self antigens that are not inducers of auto-antibodies are not chemotactic. A number of tumor antigens associated with more differentiated tumor cells such MUC1, gp100, and CEA are also chemotactic for GiPCR, while PSA and NY-ESO-1, which are perhaps more embryonic self antigens are not chemotactic. Unlike the alarmins, the chemotactic autoantigens do not activate iDC to mature to mDC. Consequently these antigens appear not to be immunogenic unless associated with a concomitant inflammatory response. However, these antigens are internalized together with the receptors by iDC and do induce the expression of some CCR7. They therefore are potentially tolerogenic. We are further characterizing the receptor interactions and signal transduction initiated by autoantigens and tumor antigens to better distinguish immunogenic from tolerogenic signals. Our auto-antigen studies expanded to include related exogenous antigen during the past year. Our collaborative studies with Dr. B.L. Ramirez et al showed that the asparaginyl-tRNA synthetase homologue produced by the Brugia Malayi parasite also has chemotactic and activating effects on human leukocytes expressing the CXCR1 and CXCR2 chemokine receptors. Infection with this organism does not lead to the development of autoimmune myositis, which is characterized by the development of antibodies to asparaginyl-tRNA synthetase. The pathophysiological relevance of the capacity of this organism to produce such a protein mimic remains to be clarified. Chemokines have been reported to promote the invasiveness and metastatic spread of cancer cells by acting on various chemokine receptors on tumor cells. We therefore pursued studies of epidermal growth factor (EGF) which in addition to promoting the growth of the many tumors that express receptors for EGF also promotes tumor spread based on its chemotactic effects. Our studies established that EGF-Receptors preferentially present in lipid rafts in the cell membrane are responsible for the chemotactic response of human breast tumor cells. The chemotactic response to EGF could be inhibited by methyl-beta-cyclodextrin and restored with cholesterol. Thus disruption of lipid rafts can potentially interfere with the trafficking of tumor cell metastases in response to EGF

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蛋白偶联趋化因子受体的相互作用，以及对炎症，自身免疫性，癌症和厌食症的影响。我们已经表明，多种抗菌肽模仿趋化因子，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。警报的特征在于表达GIPCR的白细胞趋化活性，以及​​诱导IDC成熟成抗原呈递的能力，T淋巴细胞激活树突状细胞（MDC），以及由此产生的体内免疫辅助效应。如果抗原与抗原一起施用，警报蛋白的这些活性会大大增加对抗原的细胞和体液免疫反应。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。 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单核细胞和IDC前体表达的FPRL-1受体的趋化性，并且IDC的前体诱导了IDC的成熟，并且同样像Vivo一样有效的佐剂。尽管警报在结构上是不同的，但它们迅速从白细胞颗粒中释放出来或响应角质形成细胞的促炎性刺激剂或gi gu道，gu道和气管机管树的促炎刺激剂。因此，警报可能代表预警系统，以提醒主机防御危险信号。像GM-CSF一样，像警报蛋白这样的细胞因子对树突状细胞和免疫辅助作用也具有趋化作用，这些警报蛋白可能被证明是肿瘤疫苗中的辅助剂。在本年度，我们研究了另一种白细胞颗粒衍生的催眠蛋白的免疫激活和趋化作用，称为嗜酸性神经毒素（EDN），该神经毒素（EDN）是RNase家族的成员，并具有包括抗HIV活性的抗病毒活性。 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。然而，EDN激活DC以表现出极化的Th2模式，并通过B淋巴细胞更大地诱导IgG1抗体产生，并产生白介素（IL）-5和13的产生，但没有IL-4或IFNGAMMA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，对于IDC的GIPCR，高迁移率组I（HMGB1）是具有抗病毒活性的核（DNA）结合蛋白。我们的合作者凯文·特雷西（Kevin Tracey）等人博士已显示HMGB1是促炎细胞因子的有效诱导剂，以诱导IDC对MDC的成熟，并在炎症部位高度生产并具有免疫辅助作用。因此，HMGB1是一种有效的警报蛋白，由于它诱导了诸如IL-12等细胞因子的体外产生，而IL-12又诱导了IFN Gamma，因此HMGB1作为Th1免疫反应的诱导剂可能被证明是抗肿瘤疫苗的有效佐剂。我们将测试我们已经鉴定出的一些警报蛋白是否可以将耐受性转化为免疫原性反应，这在耐受肿瘤的情况下是可取的。 GIPCR不仅与非认知警报蛋白相互作用，而且还导致IDC对许多自动抗原和某些肿瘤抗原的趋化反应。我们先前报道了组酰基TRNA合成酶（HRS）和ASNR，某些肌炎患者会出现自身抗体的自身抗原，分别是表达CCR5和CCR3受体的细胞（包括IDC）的趋化性。最近，我们证明了诱导实验性自身免疫性葡萄膜炎（EAU）的抗原，IRBP使用CXCR3和CXCR5，而S抗原仅使用CXCR3用于chemoattract IDC。与I型糖尿病，EAE和硬皮病相关的许多其他自动抗原也是IDC的趋化性。不是自身抗体诱导者的无关tRNA合成酶和自抗原不是趋化的。与更多分化的肿瘤细胞相关的许多肿瘤抗原，例如MUC1，GP100和CEA也是GIPCR的趋化性抗原，而PSA和NY-ESO-1也许是胚胎自我抗原，而不是胚胎自我抗原。与警报蛋白不同，趋化自身抗原不会激活IDC以成熟到MDC。因此，除非与伴随炎症反应相关，否则这些抗原似乎不是免疫原性。但是，这些抗原与IDC一起将其内化，并确实诱导了某些CCR7的表达。因此，它们具有潜在的耐受性。我们正在进一步表征自身抗原和肿瘤抗原引发的受体相互作用和信号转导，以更好地区分免疫原性和耐受性信号。我们的自动抗原研究扩展到了过去一年中相关的外源抗原。我们与B.L.博士的合作研究Ramirez等人表明，由Brugia Malayi寄生虫产生的天冬酰基-TRNA合成酶同源物还具有趋化性和激活对表达CXCR1和CXCR2趋化因子受体的人白细胞的作用。这种生物体的感染不会导致自身免疫性肌炎的发展，其特征在于开发与天冬酰胺基-TRNA合成酶的抗体。该生物产生这种蛋白质模仿的能力的病理生理相关性尚待澄清。据报道，趋化因子通过作用于肿瘤细胞上的各种趋化因子受体来促进癌细胞的侵袭性和转移扩散。因此，我们进行了对表皮生长因子（EGF）的研究，除了促进许多肿瘤的生长外，EGF表达受体的生长还根据其趋化作用促进肿瘤扩散。我们的研究确定，EGF受体优先存在于细胞膜中的脂质筏中，负责人乳腺肿瘤细胞的趋化反应。甲基-Beta-Cyclodextrin可以抑制对EGF的趋化反应，并用胆固醇恢复。因此，脂质筏的破坏可能会干扰响应于EGF的肿瘤细胞转移的运输