Casuarinin suppresses TARC/CCL17 and MDC/CCL22 production via blockade of NF-κB and STAT1 activation in HaCaT cells

Casuarinin is a naturally occurring tannin that is isolated from the leaves of Hippophae rhamnoides. It has been shown to have anti-oxidant, anti-cancer, anti-viral, and anti-inflammatory activities. The aim of this study was to investigate the possible mechanism by which casuarinin inhibits TNF-α/IFN-γ-induced Th2 chemokines expression in the human keratinocytes cell line HaCaT. We found that casuarinin suppressed TNF-α/IFN-γ-induced expression of TARC and MDC mRNA and protein in HaCaT cells. Casuarinin significantly inhibited TNF-α/IFN-γ-induced activation of NF-κB, STAT1, and p38 MAPK. Furthermore, we observed that p38 MAPK contributes to inhibition of TNF-α/IFN-γ-induced TARC and MDC production by blocking NF-κB and STAT1 activation in HaCaT cells. Taken together, these results suggest that casuarinin may exert anti-inflammatory responses by suppressing TNF-α/IFN-γ-induced expression of TARC and MDC via blockage of p38 MAPK activation and subsequent activation of NF-κB and STAT1. We propose that it could therefore be used as a therapeutic agent against inflammatory skin diseases.     

靶向CCL21/CCR7轴治疗肿瘤转移的研究进展

转移是肿瘤患者死亡最常见的原因,而淋巴转移是大多数肿瘤转移的主要途径之一。近年来,CC趋化因子配体21 (CC chemokine ligand 21,CCL21) 及其受体CC趋化因子受体7型 (CC chemokine receptor type 7,CCR7) 在淋巴转移中的作用逐渐受到关注。CCL21主要由淋巴内皮细胞产生,其与树突状细胞 (Dendritic cells,DCs) 和T细胞等表面CCR7的相互作用是免疫细胞淋巴迁移及淋巴结归巢的主要决定因素。然而,表达CCR7的肿瘤细胞也可以利用类似的机制进入淋巴管进行淋巴转移。如何靶向CCL21/CCR7轴,既能抑制淋巴转移,又不影响抗肿瘤免疫反应已成为肿瘤免疫治疗研究的重要议题。文中将对CCL21/CCR7轴在淋巴转移中的作用及其作为靶点治疗肿瘤转移的临床前和临床试验研究进行综述,为靶向CCL21/CCR7信号轴治疗肿瘤转移的相关研究提供参考。     

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。     

Profile of circulating levels of IL-1Ra, CXCL10/IP-10, CCL4/MIP-1β and CCL2/MCP-1 in dengue fever and parvovirosis

Dengue virus (DENV) and parvovirus B19 (B19V) infections are acute exanthematic febrile illnesses that are not easily differentiated on clinical grounds and affect the paediatric population. Patients with these acute exanthematic diseases were studied. Fever was more frequent in DENV than in B19V-infected patients. Arthritis/arthralgias with DENV infection were shown to be significantly more frequent in adults than in children. The circulating levels of interleukin (IL)-1 receptor antagonist (Ra), CXCL10/inducible protein-10 (IP-10), CCL4/macrophage inflammatory protein-1 beta and CCL2/monocyte chemotactic protein-1 (MCP-1) were determined by multiplex immunoassay in serum samples obtained from B19V (37) and DENV-infected (36) patients and from healthy individuals (7). Forward stepwise logistic regression analysis revealed that circulating CXCL10/IP-10 tends to be associated with DENV infection and that IL-1Ra was significantly associated with DENV infection. Similar analysis showed that circulating CCL2/MCP-1 tends to be associated with B19V infection. In dengue fever, increased circulating IL-1Ra may exert antipyretic actions in an effort to counteract the already increased concentrations of IL-1β, while CXCL10/IP-10 was confirmed as a strong pro-inflammatory marker. Recruitment of monocytes/macrophages and upregulation of the humoral immune response by CCL2/MCP-1 by B19V may be involved in the persistence of the infection. Children with B19V or DENV infections had levels of these cytokines similar to those of adult patients.     

CCL21/CCR7轴与乳腺癌转移及治疗的相关研究进展

乳腺癌是发生在乳腺腺上皮组织的恶性肿瘤,是全世界女性死亡的最常见原因之一。趋化因子CCL21是一种具有趋化细胞迁移功能的低分子量蛋白质,CCL21可以与淋巴细胞表面表达的CCR7受体结合,促使其迁移至乳腺肿瘤相关部位;同时还能与乳腺肿瘤细胞上表达的CCR7配体结合,使其不断向正常部位转移和侵袭,从而导致癌细胞恶性发展。CCL21/CCR7轴与乳腺癌的治疗、转移、侵袭与预后有着密切联系。我们简要综述近年来CCL21/CCR7轴与乳腺癌的转移及治疗相关研究进展,以期为乳腺癌的临床治疗提供参考。     

槐杞黄颗粒联合吸入糖皮质激素对支气管哮喘患儿肺功能、细胞免疫因子及CCL3、CCL5水平的影响

摘要 目的：探讨槐杞黄颗粒联合吸入糖皮质激素对支气管哮喘患儿肺功能、细胞免疫因子及趋化因子3（CCL3）、趋化因子5（CCL5）水平的影响。方法：按照随机数字表法,将我院收治的轻度持续性支气管哮喘患儿（n=90）,分为对照组（吸入糖皮质激素,45例）和研究组（槐杞黄颗粒联合吸入糖皮质激素,45例）,两组均治疗3个月。对比两组患儿治疗前、治疗后1、3、6个月的常规肺功能、细胞免疫因子及CCL3、CCL5水平。观察两组临床疗效、指标和不良反应。结果：研究组的临床总有效率71.11%（32/45）高于对照组91%（41/45）,差异有统计学意义（P<0.05）。治疗后1个月（T2）、治疗后3个月（T3）、治疗后6个月（T4）时间点,两组第1秒用力呼气量占预测值百分比（FEV₁%）、呼气峰流速占预测值百分比（PEF%）、最大呼气中期流速占预计值的百分比（MMEF75/25）升高,总呼吸阻抗（Z5）、肺弹性阻力（X5）及共振频率（Fres）下降,且研究组的改变程度大于对照组（P<0.05）。T2、T3、T4时间点,两组免疫球蛋白（IgE）、白细胞介素-4（IL-4）、白细胞介素-5（IL-5）、白细胞介素-13（IL-13）、白细胞介素-17（IL-17）下降,且研究组的改变程度大于对照组（P<0.05）。T2、T3、T4时间点,两组CCL3、CCL5下降,且研究组的改变程度大于对照组（P<0.05）。两组不良反应发生率组间对比无差异（P>0.05）。研究组C-ACT评分高于对照组,呼吸道感染次数、哮喘发作次数少于对照组（P<0.05）。两组因哮喘住院次数组间对比未见统计学差异（P>0.05）。结论：槐杞黄颗粒联合吸入糖皮质激素治疗支气管哮喘,可促进患儿肺功能恢复,改善细胞免疫因子及CCL3、CCL5水平。     

病毒感染对角质形成细胞CCL20表达的影响及其机制

目的:既往研究发现,趋化因子CCL20在银屑病、白癜风等在内的多种自身免疫性皮肤疾病的病理过程中扮演了重要的角色,同时病毒感染也被认为是自身免疫性疾病的重要参与者。皮肤组织是机体抵御外界病原体的第一道屏障,其中角质形成细胞被认为在启动免疫中发挥了关键的作用。视黄酸诱导基因蛋白I(RIG-I)是固有免疫模式识别受体家族的重要成员,其能够被病毒复制的中间产物激活。然而,病毒感染是否会通过RIG-I信号通路影响角质形成细胞中CCL20的表达,进而参与自身免疫性皮肤疾病的病理过程仍不清楚。本文使用聚肌胞苷酸(Poly(I:C))来体外模拟病毒感染,探究病毒感染对皮肤角质形成细胞CCL20表达的影响,并且通过小干扰RNA沉默关键分子来探究相应的分子机制。方法:首先,体外细胞实验使用Poly(I:C)刺激角质形成细胞系HaCaT,通过Western-blot实验和qRT-PCR实验探究Poly(I:C)对HaCat中RIG-I表达的影响;接下来,通过实时荧光定量PCR(qRT-PCR)以及酶联免疫吸附测定实验(ELISA)检测Poly(I:C)对角质形成细胞CCL20分泌的影响;线粒体抗病毒信号蛋白(MAVS)在RIG-I的下游发挥着重要作用,我们通过小干扰RNA(si-RNA)阻断RIG-I-MAVS-NF-κB信号通路关键分子,探究Poly(I:C)诱导角质形成细胞CCL20表达升高的分子机制。结果:Poly(I:C)能够明显促进角质形成细胞中RIG-I的表达及CCL20的表达和分泌;Poly(I:C)诱导角质形成细胞CCL20分泌是由RIG-I-MAVS-NF-κB信号通路介导的。结论:Poly(I:C)模拟病毒感染能够通过RIG-I-MAVS-NF-κB信号通路介导CCL20表达增加,进而参与自身免疫性皮肤疾病的病理过程。     

宫颈癌中CD1α分子和趋化因子CCL20/MIP-3α的表达

目的:研究CD1α分子,趋化因子人巨噬细胞炎性蛋白-3α(CCL20/MIP-3α)在宫颈癌组织中的表达及其相关性,探讨其在宫颈癌免疫逃逸机制中的作用。方法:应用免疫组化PV-6000通用二步法检测CD1α和趋化因子CCL20/MIP-3α在正常宫颈组织及宫颈癌组织中的表达,并分析两者之间的相关性。结果:CD1α和CCL20/MIP-3α在宫颈癌中的表达均明显降低(P均0.01);CD1α与CCL20/MIP-3α在宫颈癌中的表达显著相关(P0.01)。结论:在宫颈癌组织中,趋化因子的减少导致抗原提呈细胞数量下降,不足以引起肿瘤局部免疫反应而导致了宫颈局部病变的侵袭及发展。     

CCL22在人类免疫缺陷病毒感染中的作用

CCL22是CC型的趋化因子,其受体为CCR4.CCL22抑制巨噬细胞中R5人类免疫缺陷病毒1型的复制,其影响的不是病毒侵入或反转录等早期过程,而是作用于病毒侵入细胞后阶段.对CCL22独特功能的深入研究和CCR4这一病毒新共受体的发现将为人类免疫缺陷病毒的防治提供新的途径.     

CKβ8/CCL23 induces cell migration via the Gi/Go protein/PLC/PKCδ/NF-κB and is involved in inflammatory responses

AimsCKβ8/CCL23 is a CC chemokine and alternative splicing of the CKβ8 gene produces two mRNAs that encode CKβ8 and its isoform CKβ8-1. Although it has been reported that CKβ8 and CKβ8-1 are implicated in leukocyte trafficking and development of inflammation, the exact roles of these two chemokines in immune responses and the associated chemotaxis signaling are still obscure.Main methodsTo understand the mechanism of CKβ8- and CKβ8-1-induced chemotaxis signaling, we examined the chemotactic activities of osteogenic sarcoma cells expressing CC chemokine receptor 1 in response to CKβ8 and CKβ8-1. We also examined involvement of CKβ8 and CKβ8-1 in inflammatory responses by determining the mRNA expression of pro-inflammatory molecules induced by two chemokines and expressions of these chemokines in foam cells.Key findingsResults from a chemotaxis assay using various inhibitors for signaling molecules showed that the chemotaxis signal pathway induced by both CKβ8 and CKβ8-1 was mediated via the G_i/G_o protein, phospholipase C (PLC) and protein kinase Cδ (PKCδ). Treatment with a nuclear factor κB (NF-κB) inhibitor reduced the chemotactic activities of CKβ8 and CKβ8-1, and NF-κB was activated in response to CKβ8 and CKβ8-1. In addition, CKβ8 and CKβ8-1 increased mRNA expression of pro-inflammatory cytokines and adhesion molecules. The mRNA levels of CKβ8 and CKβ8-1 were increased in foam cells.SignificanceThese results indicate that both CKβ8 and CKβ8-1 transduce the chemotaxis signal through the G_i/G_o protein, PLC, PKCδ, and NF-κB, and that CKβ8 and CKβ8-1 probably play important roles in inflammatory diseases such as atherosclerosis.     

CCL2 −2518 A/G single nucleotide polymorphism as a risk factor for breast cancer

The contribution of the CCL2 −2518 A>G (rs 1024611) polymorphism in the occurrence and progression of various cancers has been found to be discordant. We studied the prevalence of the CCL2 −2518 A>G polymorphism in patients with breast cancer (n = 160) and controls (n = 323) in a sample of the Polish population. There were no significant differences in CCL2 −2518 A>G genotypes between patients with breast tumors and controls. Odds ratio (OR) for patients bearing the GG genotype was 1.481 (95% CI = 0.7711–2.845, P = 0.2358), and OR of the GG and AG genotypes was 0.7269 (95% CI = 0.4967–1.064, P = 0.1002). There was also no significant distinction in the prevalence of alleles between patients and healthy individuals. OR for the CCL2 −2518 G allele frequency was 0.8903 (95% CI = 0.6611–1.199, P = 0.4441). Analysis of the association between tumor size, lymph node metastases, histological grade, and distribution of genotypes and alleles for the CCL2 −2518 A>G polymorphism also did not show significant differences. Our results did not show association of the CCL2 −2518 A>G polymorphism with breast cancer occurrence and clinical characteristics in a sample of the Polish cohort.     

CCL27的不同剪切机制及其功能差异

CCL27是一种CC型趋化因子,其配体为CCR10。由于mRNA剪切的不同,CCL27形成两种不同的分子,即经典CCL27和其变异体PESKY,前者含分泌肽,对活化CD4＋T细胞有趋化作用,是皮肤炎症反应中扮演主要角色的趋化因子;后者含有核内定位序列,调控细胞骨架结构的重排,这一特性为该趋化因子所独有,可能预示着趋化因子家族的新功能。     

CCL20在脑卒中的发病机制及临床意义

CCL20(C-C chemokine ligand 20)是CC亚族的趋化因子,曾被称为巨噬细胞炎症蛋白3α(MIP-3α)、肝脏活化调节趋化因子(LARC)和Exodus-1,与其唯一受体CC趋化因子受体6(CC chemokine receptor 6,CCR6)特异性结合,调节细胞的激活、趋化和迁移,参与形态发生、组织稳态、固有和获得性免疫以及许多疾病的炎症病理过程。在中枢神经中,CCL20与CCR6结合后激活小胶质细胞,其可应激相关性蛋白,形成持续性神经毒性级联反应,从而进一步损害周围神经元和脑组织,进而引起一系列的神经功能损伤症状。目前有关CCL20在脑卒中的日益引起人们的关注,深入研究其在脑卒中的作用,有可能为进一步揭示脑卒中的发病机制并为寻求有效的防治措施提供重要的理论基础。本文概述了CCL20的作用,重点阐述了其在脑卒中的作用。     

人CCL20基因打靶载体的构建与鉴定

目的:构建针对人CC亚族趋化因子配体20(CC chemokine ligand 20,CCL20)基因外显子2的置换型打靶栽体.方法:设计和合成引物,利用长距离聚合酶链反应(polymerase chain reaction,PCR)从永生化人角质形成细胞系(HaCaT)基因组DNA中克隆出CCL20基因组DNA片段,再以CCL20基因为模板扩增出长度为1969 bp的同源短臂和2356 bp的同源长臂.分别插入pioxP栽体的Neo基因上游和下游,构建针对人CCL20基因外显子2的置换型打靶载体(ploxP-hCCL20).将打靶载体线性化并以电穿孔方法转移入HaCaT,观察克隆的形成.结果:经过PCR、限制性内切酶鉴定及DNA序列测定.证实该载体的两条同源臂包含人CCL20基因的外显子1、3、4及其邻近的部分内含子.结论:ploxP-hCCL20载体构建成功,增加Neo基因下游同源臂长度的策略有可能提高细胞基因敲除的同源重组率.     

Local production of CCL3, CCL11, and IFN-γ correlates with disease severity in murine parainfluenza virus infection

Background

Using a murine model of parainfluenza virus infection (mPIV1 or Sendai virus; SeV), we compared the inflammatory responses to lethal and sub-lethal infections in inbred DBA/2 mice.

Methods

Mice were intranasally inoculated with either 1.6×10³ or 1.6×10⁵ infectious units (IU) of SeV or diluent control. Clinical data including daily weights, oxygen saturation, and lung function via whole body plethysmography were collected on days 0, 3–7, and 9–14. Clarified whole lung homogenates were evaluated for inflammatory markers by enzyme-linked immunoassay (ELISA). Data were analyzed using ANOVA or Student t-tests, as appropriate.

Results

Mice inoculated with 1.6×10⁵ IU of SeV developed a lethal infection with 100% mortality by day 7, while mice inoculated with 1.6×10³ IU developed a clinically significant infection, with universal weight loss but only 32% mortality. Interestingly, peak virus recovery from the lungs of mice inoculated with 1.6×10⁵ IU of SeV did not differ substantially from that detected in mice that received the 100-fold lower inoculum. In contrast, concentrations of CCL5 (RANTES), CCL11 (eotaxin), interferon-γ, CXCL10 (IP-10), and CCL3 (MIP-1α) were significantly higher in lung tissue homogenates from mice inoculated with 1.6×10⁵ IU (p?<?0.05). In the lethal infection, levels of CCL11, interferon- γ and CCL3 all correlated strongly with disease severity.

Conclusion

We observed that severity of SeV-infection in DBA/2 mice was not associated with virus recovery but rather with the levels of proinflammatory cytokines, specifically CCL11, interferon- γ and CCL3, detected in lung tissue in response to SeV infection.

     

CCL27及其受体在抗病毒免疫中的作用

趋化因子是一类结构相关、对白细胞具有趋化活性、相对分子质量为(8～10)×103的小分子蛋白质,在炎症和免疫反应中可诱导白细胞和淋巴细胞迁移.CCL27是一种CC型趋化因子,其配体为CCR10.CCL27与接触性软疣病毒编码产生的蛋白质MC148有较高的同源性,但却与人疱疹病毒8型编码产生病毒巨噬细胞炎症蛋白-Ⅱ竞争结合受体CCR10.     

CCL2促进大鼠原代心肌微血管内皮细胞血管形成

本文旨在探讨趋化因子CCL2在成年大鼠原代心肌微血管内皮细胞(cardiac microvascular endothelial cell, CMEC)血管形成中的作用。分离原代大鼠CMEC,用CD31和VIII因子免疫染色鉴定,观察不同时间点基质胶(Matrigel)上CMEC血管形成情况,用real-time RT-PCR和ELISA分别检测在血管形成过程中CCL2的表达和分泌情况。结果显示:(1)大鼠原代CMEC分离成功,该细胞具有形成血管能力,成网数和节点数在Matrigel处理后4 h显著增加;(2) CMEC血管形成过程中CCL2和CCR2的表达量显著上调,并且CCL2的分泌量明显增加;(3) CCL2阻断抗体和CCR2拮抗剂均可显著降低CMEC血管形成能力。上述结果提示,大鼠原代CMEC在血管形成过程中可分泌CCL2,并且CCL2-CCR2信号通路促进了CMEC血管形成。     

CCL4在病毒性心肌炎中的作用及其机制研究

目的:观察巨噬细胞炎性蛋白-1β(MIP-1β/CCL4)在病毒性心肌炎小鼠血清以及心肌组织中的变化,初步探讨CCL4在病毒性心肌炎中发挥的作用。方法:雄性Balb/c小鼠随机分成对照组15只和病毒性心肌炎组25只,病毒性心肌炎组腹腔注射病毒液,对照组腹腔注射Eagle's培养液。15 d处死小鼠,通过HE染色观察小鼠心肌病理的变化;RT-PCR观察CCL4以及促炎因子的m RNA水平,Western blotting观察CCL4蛋白水平的变化。结果:与正常对照组比,病毒性心肌炎组小鼠生存质量降低,生存率也显著降低;病毒性心肌炎组小鼠心肌炎性浸润严重,心肌组织中的CCL4、IL-1β、TNF-α mRNA水平显著增高,CCL4蛋白水平也明显增高,并且CCL4的浓度和蛋白表达水平与心肌病变积分呈正相关。结论:病毒性心肌炎CCL4的表达显著增加,并且与心肌病变程度相关。