CCL20/CCR6 promotes the invasion and migration of thyroid cancer cells via NF-kappa B signaling-induced MMP-3 production

CCL20, an important member of the CC-chemokine family, is the only ligand that activates CCR6. The levels of CCL20 and CCR6 are elevated in many human cancers, and CCL20/CCR6 interaction participates in the development and progression of cancer. In this present study, we found that CCR6 was overexpressed in thyroid cancer cells. Activation of CCR6 by CCL20 promoted the invasion and migration of human thyroid cancer SW1736 cells, while knockdown of CCR6 repressed the effect of CCL20. Furthermore, CCL20/CCR6 interaction induced the activation of NF-κB, and stimulated the expression and secretion of MMP-3. In addition, BAY117082, a special inhibitor of NF-κB, suppressed the expression and secretion of MMP-3 stimulated by CCL20/CCR6. Together, these results suggest that CCL20/CCR6 enhances thyroid cancer cell invasion and migration. The possible molecular mechanisms involved NF-κB activation and NF-κB-dependent MMP-3 upregulation. Thus, molecular therapies that aim at CCL20 and CCR6 may offer promising intervention strategies for thyroid cancer.     

Elevated CCR6+ CD4+ T lymphocytes in tissue compared with blood and induction of CCL20 during the asthmatic late response

CCR6 is expressed by multiple leucocyte subsets, including peripheral blood memory T cells, and mouse models implicate a role for this receptor in diverse inflammatory responses that include allergic airway disorders, inflammatory bowel disease and autoimmune encephalitis. In order to study the role of CCR6 in humans, we have investigated the patterns of CCR6 expression and function on T cells from the peripheral blood, skin, nose and lung, in health and in allergic disease. Results show that CCR6 was expressed consistently on a higher proportion of tissue versus peripheral blood-derived CD4+ T cells (P < 0.01). CCR6 was expressed predominantly on CD4+ compared with CD8+ cells in both blood- and tissue-derived T cells (P < 0.001). The number of cells showing CCR6 expression was not proportionally greater in peripheral blood or nasal mucosal T cells of subjects with symptomatic allergic rhinitis. CCR6+ cells demonstrated enhanced functional responses to CCL20 and CCL20 was increased in bronchoalveolar lavage fluid of asthmatics following endobronchial allergen provocation (P < 0.05). Thus, CCR6 may be important in the regulation of T cell recruitment to tissue and up-regulation of CCL20 expression may contribute to the recruitment and/or retention of effector T cells in allergic asthma.     

The CCR6-CCL20 axis in humoral immunity and T-B cell immunobiology

Traditionally, chemokine immunobiology has focused on chemotaxis and the positioning of cells at sites of inflammation and within lymphoid organs. More recently, however, regulation of intricate immune responses has emerged as a function attributed to chemokines and their receptors. One such pair, CCR6 and its chemokine ligand CCL20, has been receiving interest for its potential role in the coordination and regulation of humoral immune responses and in particular, memory responses, at the cellular level. B cells up-regulate CCR6 after activation in secondary lymphoid organs; however, its function is still unclear. In an important insight, the CCR6-CCL20 chemokine axis has been implicated in the regulation of effective humoral responses – disruption of this pair led to an increased number of ineffective T-B cell conjugates and poorer quality antibodies. Interestingly, follicular helper T cells and their precursors also up-regulate CCR6; though, again, the precise purpose of this is yet to be discovered. The chemokine axis in relation to secondary lymphoid organ (SLO) structures will be briefly reviewed as well. With the implication of CCR6 and CCL20 in the pathogenesis of autoantibody-driven autoimmune diseases such as systemic lupus erythematosus, understanding the intricacies of this chemokine pair would be conducive to the development of appropriate, targeted therapeutic strategies.     

In situ expression of the CCL20-CCR6 axis in lymphocyte-rich gastric cancer and its potential role in the formation of lymphoid stroma

Lymphocyte-rich gastric cancer (Ly-rich GC) is characterized by lymphoid stroma. To understand its formation, we studied the expression of a chemokine ligand (CCL)20 and its receptor CCR6 in 36 and 37 cases of Ly-rich- and conventional GC, respectively. Lymphoid tissues in the alimentary tract were studied in parallel. By quantitative polymerase chain reaction, Ly-rich GC contained CCL20 and CCR6 mRNAs at higher levels than conventional GC. By immunohistochemistry, CCL20 was expressed by cancer cells more frequently in Ly-rich GC than in conventional GC. This was comparable with its expression in epithelial cells of the alimentary tract lymphoid tissues. CCR6 was mostly expressed by dendritic cells (DC) and B cells in Ly-rich GC, which was also comparable with its expression in the alimentary tract lymphoid tissues. Cancer cells also expressed CCR6. However, its expression did not differ between Ly-rich- and conventional GC, nor was it related to the stage of cancer. Given that the CCL20-CCR6 axis is involved in the formation of alimentary tract lymphoid tissue, the similarity between the lymphoid stroma of Ly-rich GC and the alimentary tract lymphoid tissues supports the notion that it plays a significant role in the formation of lymphoid stroma in Ly-rich GC.     

IL-23/IL-17轴在银屑病免疫发病机制中的作用

银屑病是一种以T淋巴细胞异常活化和浸润为主要特征的慢性炎性反应皮肤病。Th17细胞及IL-23/IL-17轴在银屑病的发病机制中可能处于关键地位,并成为新的治疗靶标。IL-23诱导Th17细胞分化增殖,分化成熟的Th17可以分泌IL-17、IL-21、IL-22等多种细胞因子,Th17类细胞因子在银屑病等多种自身免疫疾病和炎性疾病中起重要作用。     

The airway smooth muscle CCR3/CCL11 axis is inhibited by mast cells

Background:  Airway smooth muscle hyperplasia is a feature of asthma, and increases with disease severity. CCR3-mediated recruitment of airway smooth muscle progenitors towards the airway smooth muscle bundle has been proposed as one possible mechanism involved in airway smooth muscle hyperplasia. Mast cells are microlocalized to the airway smooth muscle bundle and whether mast cells influence CCR3-mediated migration is uncertain.
Methods:  We examined the expression of CCR3 by primary cultures of airway smooth muscle cells from asthmatics and nonasthmatics. CCR3 function was examined using intracellular calcium measurements, chemotaxis, wound healing, cell proliferation and survival assays. We investigated the recovery and function of both recombinant and airway smooth muscle-derived CCL11 (eotaxin) after co-culture with β-tryptase and human lung mast cells.
Results:  Airway smooth muscle expressed CCR3. Airway smooth muscle CCR3 activation by CCL11 mediated intracellular calcium elevation, concentration-dependent migration and wound healing, but had no effect on proliferation or survival. Co-culture with β-tryptase or mast cells degraded recombinant and airway smooth muscle-derived CCL11, and β-tryptase inhibited CCL11-mediated airway smooth muscle migration.
Conclusions:  CCL11 mediates airway smooth muscle migration. However co-culture with β-tryptase or mast cells degraded recombinant and airway smooth muscle-derived CCL11 and inhibited CCL11-mediated airway smooth muscle migration. Therefore these findings cast doubt on the importance of the CCL11/CCR3 axis in the development of airway smooth muscle hyperplasia in asthma.     

GM-CSF-producing CCR2+CCR6+ Th17 cells are pathogenic in dextran sodium sulfate-induced colitis model in mice

Although excessive immune responses by Th17 cells, a helper T cell subset, are implicated in the pathogenesis of inflammatory bowel disease (IBD), the mechanism by which its localization in an inflamed colon is regulated remains unclear. Chemokines and their receptors are involved in the pathogenesis of IBD, however, the relative significance of each receptor on Th17 cells remains unknown. We generated C–C motif chemokine receptor 2 (CCR2) knockout (KO) and CCR6 KO mice in the syngeneic background using the CRISPR/Cas9 system and found that the phenotypes of experimental colitis worsened in both mutant mice. Surprisingly, the phenotype of colitis in CCR2/CCR6-double knockout (CCR2/6 DKO) mice was opposite to that of the single-deficient mice, with significantly milder experimental colitis (p < .05). The same was true for the symptoms in CCR6 KO mice, but not in wild type mice treated with a CCR2 inhibitor, propagermanium. Colonic CCR2⁺CCR6⁺ Th17 cells produced a potentially pathogenic cytokine GM-CSF whose levels in the gut were significantly reduced in CCR2/6 DKO mice (p < .05). These results suggest that GM-CSF-producing CCR2⁺CCR6⁺ Th17 cells are pathogenic and are attracted to the inflamed colon by either CCR2 or CCR6 gradient, which subsequently exacerbates experimental colitis in mice.     

CCR2、 CCL5、 CCR5和CCL1基因多态性与中国汉族儿童结核病易感性的关联研究

目的 探讨CCR2、CCL5、CCR5和CCL1的基因多态性与中国汉族儿童结核病易感性的关系.方法 收集353例汉族儿童结核病患者,以同期查体的400名儿童作为对照,采用病例对照研究,应用高通量MassARRAY技术对于CCR2、CCL5、CCR5和CCL1基因的SNP位点进行基因分型研究.结果 CCR2、CCL5、CCR5和CCL1基因SNP位点的等位基因、基因型以及单体型在结核病组和对照组的分布差异均无统计学意义（P＞0.05）.结论 CCR2、CCL5、CCR5和CCL1的基因多态性与中国汉族儿童结核病易感不存在相关性.     

CCL17 and CCL22 attenuate CCL5-induced mast cell migration

BACKGROUND: Mast cells (MCs) accumulate at sites of allergic mucosal inflammation where they act as central effectors and regulatory cells. Chemokines are believed to be crucial for the recruitment of MCs to sites of inflammation. We recently reported that human umbilical cord blood MCs (CBMCs) expresses the CC chemokine receptors, CCR1 and CCR4. We found a unique response profile to ligands of the respective receptors in which, of all tested ligands, only CCL5/RANTES-induced migration. OBJECTIVE: To further investigate the function of CCR4 in MCs. METHODS: CBMCs were used for competition binding experiments, migration, and intracellular calcium mobilization and release response studies. RESULTS: The natural ligands for CCR4, CCL17/TARC and CCL22/MDC could both compete for binding with radiolabelled CCL5. Further, both CCL17 and CCL22 act as CCR4 antagonists by inhibiting CCL5-induced migration. Although both CCL17 and CCL22 caused mobilization of intracellular calcium, none of them induced migration or histamine release. CONCLUSIONS: These results suggest that CCL5-induced migration of MCs via CCR4 can be regulated by the natural agonists CCL17 and CCL22, which are up-regulated at sites of allergic inflammation.     

凉血解毒汤对银屑病小鼠皮肤组织CCL20/CCR6表达的影响

目的:观察凉血解毒汤对银屑病小鼠皮肤组织中CC趋化因子配体20(CCL20)/CC趋化因子受体6(CCR6)表达的干预作用。方法:24只BALB/c雄性小鼠随机分为空白对照组、银屑病模型组、凉血解毒汤中药治疗组和趋化因子CCL20单克隆抗体(阳性药)治疗组。用咪喹莫特诱导小鼠银屑病模型,采用银屑病皮损面积和疾病严重程度(psoriasis area and severity index,PASI)评分标准观察银屑病样小鼠皮损的变化情况。光镜下观察皮损组织形态学变化,测量表皮层厚度。采用real-time PCR检小鼠皮肤组织样本中CCL20和CCR6表达的变化。结果:银屑病模型组小鼠皮肤出现大量鳞屑和红斑,表皮增厚;与银屑病模型组小鼠比较,凉血解毒汤组小鼠银屑病样皮损程度较轻,红斑、鳞屑以及表皮增厚程度轻于模型组,PASI分数降低,皮肤组织CCL20以及CCR6的表达明显低于模型组。趋化因子CCL20单克隆抗体治疗组小鼠银屑病皮损程度较轻,红斑、鳞屑以及表皮增厚程度轻于模型组,PASI分数降低,皮肤组织CCL20以及CCR6的表达明显低于模型组。结论:凉血解毒汤可能通过下调CCL20/CCR6的表达来减轻银屑病小鼠的皮肤病变。     

CCR10 and CCL27 are overexpressed in cutaneous squamous cell carcinoma

C-C chemokine receptor (CCR)10 is a specific receptor for chemokine ligand (CCL)27, a selective chemoattractant for skin-associated memory T cells to cutaneous sites. In melanoma, CCR10 increases the ability of neoplastic cells to grow, invade tissues, disseminate to lymph nodes, and escape the host immune responses. In this study, we investigated the expression of CCR10 and its ligand CCL27 in squamous cell carcinoma (SCC). CCR10 and CCL27 were expressed in SCC, actinic keratosis (AK), Bowen's disease, and seborrheic keratosis (predominantly prickle cell type), but not in seborrheic keratosis (predominantly basal cell type) and basal cell carcinoma. Furthermore, CCR10 and CCL27 were overexpressed in SCC relative to Bowen's disease, an early stage of SCC. Consistently, a human SCC cell line, A253 cells, and HaCaT cells exhibited CCL27 production that was strongly induced by tumor necrosis factor-α and interleukin-1β. Finally, A253 cells expressed stronger intracellular CCR10 compared to HaCaT cells by flow cytometry. These results suggest that CCR10 and CCL27 overexpression in SCC is related to the progression of SCC and is useful for the diagnosis of SCC.     

Up-regulation of CCL17, CCL22 and CCR4 in drug-induced maculopapular exanthema

BACKGROUND: Maculopapular exanthema has been reported to be the most frequently drug-induced cutaneous reaction. Although T lymphocytes are involved in the pathomechanism of this disease, little is know about the recruitment of these cells to the skin. OBJECTIVE: The aim of this work is to study the role of the chemokines TARC/CCL17 and MDC/CCL22 in the lymphocyte trafficking to affected skin in drug-induced exanthemas. METHODS: Real-time PCR was performed to quantify gene expression levels of CCL17, CCL22 and their receptor CCR4 in lesional skin biopsies and in peripheral blood mononuclear cells from patients. CCL27 and CCL22 proteins were detected in the skin by immunochemistry. Protein expression of CCR4 was determined by flow cytometry in peripheral blood lymphocytes. Functional migration assays to CCL17 and CCL22 were assessed to compare the migratory responses of peripheral blood lymphocytes from patients and healthy subjects. RESULTS: CCL17 and CCL22 were up-regulated in maculopapular exanthema-affected skin. CCR4 mRNA levels and protein expression were increased in peripheral blood mononuclear cells during the acute phase of the disease. The increased expression of the receptor was consistent with a higher response of peripheral blood lymphocytes to CCL17 and CCL22 compared with the migratory response in healthy donors. CONCLUSION: TARC/CCL17 and MDC/CCL22 might cooperate in attracting T lymphocytes to skin in drug-induced maculopapular exanthemas.     

Chemokines and common variable immunodeficiency; possible contribution of CCL19, CCL21 and CCR7 to immune dysregulation

Common variable immunodeficiency (CVID) is a heterogeneous syndrome characterized by defective immunoglobulin production and high frequency of bacterial infections, autoimmunity and manifestations of chronic inflammation. The homeostatic chemokines CCL19 and CCL21 and their receptor CCR7 are associated with modulation of inflammatory responses. CVID patients have decreased proportions of CCR7⁺ T cells in peripheral blood and we hypothesized a further dysregulation of CCL19/CCL21/CCR7 in CVID. Serum levels of CCL19 and CCL21 were compared in CVID patients and controls. T cell expression of CCR7 was related to clinical characteristics in CVID patients. Spleens extirpated from CVID patients were analysed for expression of CCL19, CCL21 and CCR7. Peripheral blood mononuclear cells (PBMC) from CVID patients and controls were analysed for cytokine response on stimulation with CCL19 and CCL21. The main findings were: (i) CVID patients have raised serum levels of CCL19 and CCL21 independently of features of chronic inflammation; (ii) CCL19 and CCR7 have similar expression in spleens from CVID patients and controls, while CCL21 is variably down‐regulated in spleens from patients; (iii) T cell expression of CCR7 is particularly low in patients characterized by chronic inflammation in vivo; and (iv) PBMC from CVID patients had attenuated cytokine response to stimulation with CCL19 and CCL21. CVID patients have raised circulatory levels of CCL19 and CCL21, and an attenuated cytokine response to stimulation with these chemokines. Because CCR7, CCL19 and CCL21 are key mediators balancing immunity and tolerance in the immune system, the abnormalities of these mediators might contribute to the profound immune dysregulation seen in CVID.     

CCL25和CCL28及其相应受体CCR9和CCR10

CCL25和CCL28是CC趋化因子家族成员,主要表达于胸腺树突状细胞和黏膜上皮细胞,CCL25和CCL28相应的受体分别是CCR9和CCR10,表达于T淋巴细胞和B淋巴细胞。CCL25和CCL28及其相应受体对循环IgA浆母细胞和T淋巴细胞的归巢起重要作用,而且CCL28还具有广谱的抗微生物活性。     

Angiogenesis drives psoriasis pathogenesis

Psoriasis pathogenesis is closely associated with disease-inducing Th1 and Th17 cells. Yet, several studies suggest that aberrant keratinocyte or endothelial cell signalling significantly contributes to disease manifestation. Histological hallmarks of psoriatic skin include the infiltration of multiple immune cells, keratinocyte proliferation and increased dermal vascularity. Formation of new blood vessels starts with early psoriatic changes and disappears with disease clearance. Several angiogenic mediators like vascular endothelial growth factor, hypoxia-inducible factors, angiopoietins and pro-angiogenic cytokines, such as tumour necrosis factor (TNF), interleukin (IL)-8 and IL-17, are up-regulated in psoriasis development. Contact- and mediator-dependent factors derived from keratinocytes, mast cells and immune cells may contribute to the strong blood vessel formation of psoriasis. New technologies and experimental models provide new insights into the role of angiogenesis in psoriasis pathogenesis. Interestingly, many therapies target not only immune cells, but also protein structures of endothelial cells. Here we summarize the role of pro-angiogenic factors in psoriasis development and discuss angiogenesis as a potential target of novel therapies.     

CCR9/CCL25介导小鼠经生殖道hCGβ避孕疫苗粘膜免疫后B细胞归巢

目的：探讨hCGβ粘膜避孕疫苗体液免疫的分子机制。方法：分别用1010个重组乳酸杆菌Lb.pIlac、Lb.pIlac-hCGβ经小鼠生殖道粘膜途径免疫6～8周龄BALB/c小鼠,3周后相同剂量加强免疫1次。间接ELISA检测加强免疫后2周阴道灌洗液中抗hCGβIgG和IgA抗体滴度;ELISPOT检测分泌抗hCGβIgG和IgA抗体分泌细胞数;RT-PCR筛查小鼠子宫B细胞上18种趋化因子受体和相关趋化因子CCL25;FCM和ELISA分析CCR9/CCL25的表达。结果：经小鼠生殖道粘膜接种1010Lb.pIlac-hCGβ可在生殖道局部诱导产生抗hCGβIgG和IgA抗体,且以IgG抗体为主;局部存在抗hCGβIgG和IgA抗体分泌细胞,而以IgG抗体分泌细胞为主;粘膜免疫后,小鼠生殖道局部B细胞上CCR9表达增高,粘膜局部相应配体CCL25表达亦增高。结论：重组乳酸杆菌活疫苗Lb.pIlac-hCGβ经小鼠生殖道粘膜免疫可诱导有效的免疫应答;粘膜局部CCR9/CCL25可介导B细胞归巢至生殖道局部,这可能增强hCGβ粘膜避孕疫苗的避孕效果。     

趋化性细胞因子CCL28的功能与表达

CCL28是新近发现的趋化性细胞因子家族CC类中的成员。多种粘膜组织的上皮细胞可以产生CCL28,其受体为CCR10和CCR3。通过与相应受体结合,CCL28可介导血循环中的CD4^＋T细胞、CD8^＋T细胞、IgA抗体分泌细胞（ASC）等免疫活性细胞归巢迁徙至相关组织;另外,CCL28还具有明显的广谱抗微生物活性。因而,在粘膜免疫及某些肿瘤的免疫中,CCL28发挥着重要作用。CCL28的表达受到多种内外因素的影响。对其表达调节机制的阐明,必将为相关疾病的防治研究提供新的思路。     

Expression and role of CCR6/CCL20 chemokine axis in pulmonary sarcoidosis

We have shown previously that the chemokine receptors CXCR3 and CXCR6 are coexpressed by Th1 cells infiltrating the lung and the granuloma of patients with sarcoidosis. In this study, we evaluated the role of CCL20/CCR6 interaction in the pathogenesis of acute and chronic pulmonary sarcoidosis. By flow cytometry and molecular analyses, we have demonstrated that Th1 cells isolated from the bronchoalveolar lavage (BAL) of patients with sarcoidosis and T cell alveolitis are equipped with CCR6. Furthermore, CCR6(+) T cells coexpressed the chemokine receptors CXCR3 and CXCR6. Immunohistochemical analysis of lung specimens has shown that CCR6(+) T cells infiltrate lung interstitium and surround the central core of the granuloma. It is interesting that CCR6 was never detected on the alveolar macrophage (AM) surface, and it is observed in the cytoplasm of AMs from patients with sarcoidosis and alveolitis. The CCR6 ligand CCL20 was expressed by macrophages, multinucleated giant cells, and epithelioid cells infiltrating the granuloma. Furthermore, detectable levels of CCL20 protein are seen in the BAL fluid components of patients with active sarcoidosis, and sarcoid AMs release the CCR6 ligand in vitro. From a functional point of view, sarcoid Th1 cells were able to respond to CXCL10, CXCL16, and CCL20 in migratory assays. In vitro kinetic studies demonstrated that CCR6 is induced rapidly by IL-2, IL-18, and IFN-gamma. In conclusion, T cells expressing CCR6, CXCR3, and CXCR6 act coordinately with respective ligands and Th1 inflammatory cytokines in the alveolitic/granuloma phases of the disease.