辅助受体CCR5、CCR2及细胞趋化因子SDF1基因多态性与HIV-1异性传播的关系

目的 进一步阐述CCR5、CCR2和SDFl基因多态性与HIV-1异性传播的关系。方法 通过PCR／RFLP技术分析CCR5、CCR2和SDF1基因的多态性，继而分析基因多态性与HIV-1异性传播的关系。结果 在收集到的70对异性配对病例中，未能在汉族人群发现CCR5基因缺失突变，维吾尔族人CCR5Δ32基因频率为6．5％(6／92)，未发现纯合突变。有暴露史而未感染HIV-1者CCR2-64I基因频率明显高于受暴露后感染病毒者，说明CCR2-64I对异性传播可能具有一定保护作用。对SDFl基因的多态性研究发现，将病毒传播给配偶的指标病例(先感染：HIV-1一方)SDF-3′A的基因频率高于未发生传播者(较接近于统计学检验水准)，SDFl-3′A似乎是危险因素。结论 CCR5Δ32对汉族人群的HIV-1异性传播无明显意义，CCR2-64I对HIV-1异性传播可能具有一定的保护作用，而SDFl-3′A则有可能是危险因素，但有必要扩大样本量对后二者作进一步的深入研究。     

趋化因子CCL28及其受体CCR10的分子生物学

CCL28是趋化因子CC家族成员,其受体为CCR10和CCR3.CCL28与CC家族其他成员有同源性,但是它在基因定位、蛋白质结构和表达调控上有其独特性,这决定了它不仅可以与CCR10结合诱导循环IgA浆母细胞的归巢和迁移,而且具有广谱的抗微生物活性.     

安徽省汉族SLE患者和正常人RANTES单核苷酸多态性及CCR5基因多态性的比较

目的　探讨中国汉族系统性红斑狼疮 (SLE)患者和正常人群RANTES单核苷酸多态性(SNP)及其受体CCR5多态性。方法　收集 14 6例确诊的SLE患者和 15 9名正常对照。通过PCR RFLP方法检测研究对象RANTES启动区SNP及其受体CCR5△ 32突变频率。结果　病例组RANTES 4 0 3位点G G、G A、A A基因型频率分别为 76 .71%、2 1.92 %、1.37% ;对照组分别为 6 7.30 %、2 9.5 6 %和3.14 % ,两组间基因型分布差异无显著性 (P >0 .0 5 )。两组突变等位基因 4 0 3A频率分别为 12 .3%、17.9% (P >0 .0 5 )。病例组RANTES 2 8位点C C、C G、G G基因型频率分别为 93.15 %、6 .85 %、0 ;对照组分别为 86 .79%、12 .5 8%和 0 .6 3% (P >0 .0 5 )。两组突变等位基因 2 8G频率分别为 3.4 %、6 .9% (P>0 .0 5 )。病例组和对照组突变等位基因CCR5△ 32频率分别为 0、0 .3% (P >0 .0 5 )。中国汉族人群RANTES突变基因型 4 0 3A A低于北美黑人和西非黑人 (P <0 .0 5 ) ,与北美高加索、北美西班牙人和北美亚洲人一致。RANTES 2 8位点基因型分布与北美亚洲人一致 ,但与北美高加索、北美西班牙人、北美黑人和西非黑人相差较大 (P <0 .0 5 )。结论　本次研究发现RANTES 4 0 3位点和 2 8位点单核苷酸多态性与SLE发病没有直接的关系。CCR5△ 32     

CCR5△32、CCR5m303、CCR2-64I、SDF1-3''A基因多态性对中国HIV-1感染者预后的影响

目的研究CCR5A32、CCR5m303、CCR2-64I、SDF1—3’A基因多态性对中国HIV-1感染者预后的影响。方法对在深圳地区发现的HIV-1感染者进行流行病学调查，应用PCR／RFLP技术分析感染者CCR5G32、CCR5m303、CCR2-64I、SDF1-3’A4种基因的多态性，对部分人群进行HIV-1血浆病毒载量和CD4^＋细胞计数的检测，判断感染者的潜伏期，使用SPSS11．0统计软件分析基因多态性对感染者病毒载量和潜伏期的影响。结果在189例HIV-1感染者中没有发现CCR5A32和CCR5-m303突变基因型，SDF1—3’A的等位基因频率为26．14％，CCR2—64I的等位基因频率为19．82％。方差分析发现，CCR2-64I基因野生型与杂合型的两组HIV-1感染者人群病毒载量对数的大小差异无统计学意义（P=0．272），两组人群潜伏期的长短差异无统计学意义（P=0．662）。SDF1基因野牛型、杂合型和纯合型的3组HIV-1感染者人群病毒载量对数的大小差异有统计学意义（P=0．001），但3组人群潜伏期的长短差异无统计学意义（P=0．228）。结论CCR2-64I基因突变对中国汉族HIV-1感染者病毒载量没有明显影响，因而也不影响感染者的潜伏期。SDF1-3’A基因突变对于病毒载量有降低作用，但对延长感染者潜伏期可能没有作用。     

HSP70/CD80 DNA疫苗通过调节趋化因子及受体对哮喘小鼠气道炎症的影响

目的 观察HSP70/CD80 DNA疫苗通过调节趋化因子及趋化因子受体对急性哮喘小鼠气道炎症和气道高反应性的治疗作用.方法 以卵清蛋白(OVA)致敏小鼠,建立小鼠急性哮喘模型,同时肌肉注射HSP70/CD80 DNA疫苗.观察小鼠气道反应性,外周血IgE,肺组织炎症和黏液分泌情况,CCL5和CCL17在气道表达情况.Real-time PCR检测肺组织CCR5和CCR4基因表达.结果 与对照小鼠相比,急性哮喘小鼠的气道反应性明显增高,血清IgE含量明显升高(P＜0.05),肺组织炎细胞浸润明显(P＜0.05),大量黏液形成.HSP70/CD80 DNA疫苗治疗小鼠后,能有效减轻气道高反应性,降低血清IgE含量,降低抑制气道炎细胞浸润,减少黏液分泌(P＜0.05);且CCL5在气道上皮呈阳性表达,CCL17呈阴性(P＜0.05);肺组织CCR5基因表达增加和CCR4表达减少(P＜0.05).结论 HSP70/CD80 DNA疫苗可通过增强CCL5在气道上皮中的表达,减少CCL17表达;上调CCR5抑制CCR4,使CCR5/CCR4升高,从而恢复Th 1/Th2平衡,起到治疗哮喘的作用.     

ABCB1和ABCC2基因多态性与中国汉族儿童抗结核药物致肝毒性易感性的相关性研究

目的 探讨ABCB1和ABCC2基因多态性与中国汉族儿童抗结核药物致肝毒性(anti-tuberculosis drug-induced hepatotoxicity,ATDH)易感性的相关性.方法 在中国汉族结核病患儿中,采用病例对照研究,利用高通量的MassARRAY平台,对于ABCB1和ABCC2基因的16个标签单核苷酸多态性(single nucleotide polymorphisms,SNPs)位点展开基因分型,并采用logistic回归分析以上SNP位点等位基因和基因型频率在ATDH病例组和ATDH对照组中的分布差异.结果 本研究共纳入41例ATDH病例以及189例对照.ABCB1和ABCC2基因SNP位点的等位基因以及基因型在两组间频率分布差异均无统计学意义(P＞0.05).SNP位点分别按照显性遗传模型和隐性遗传模型分析,各位点基因型在两组间频率分布差异均无统计学意义(P＞0.05).结论 ABCB1和ABCC2基因多态性可能与中国汉族儿童ATDH的发生并无相关性.     

PIK3CA、CCL5、Survivin基因与甲状腺癌相关性的新发现

三种基因PIK3CA、CCL5、Survivin参与多种疾病过程,尤其与癌症关系密切.研究表明,PIK3CA、CCL5、Survivin基因多态性在甲状腺癌的发生、浸润、转移、治疗和预后方面发挥重要作用.     

早孕期人蜕膜趋化因子CCL2及其受体CCR2的表达及意义

目的：分析人早孕蜕膜及蜕膜基质细胞趋化因子受体CCR2及其配体CCL2在人早孕蜕膜组织及蜕膜基质细胞的表达和分泌，以探讨CCR2／CCL2在母-胎界面的生物学作用。方法：收集早孕期蜕膜组织，分离蜕膜基质细胞，分别用半定量RT-PCR、免疫化学方法分析正常人早孕蜕膜组织和培养的人蜕膜基质细胞CCR2／CCL2表达；并且用流式细胞术和ELISA法分别检测蜕膜基质细胞表面CCR2的表达和培养的蜕膜基质细胞上清中CCL2的分泌。结果：人早孕蜕膜组织和蜕膜基质细胞均高水平转录和翻译CCR2／CCL2，培养的基质细胞能分泌大量的CCL2，其分泌量呈时间依赖性。结论：早孕蜕膜高表达和分泌CCR2／CCL2可能参与早孕期母一胎免疫调节。     

CCR5基因启动子59029位点单核苷酸多态性与HIV-1疾病进展关系的Meta分析

目的探讨CCR5基因启动子59029位点单核苷酸多态性（SNPs）与HIV-1感染者疾病进展的关系。方法按照制定的检索策略,在PubMed、CBM、VIP等数据库检索有关CCR5基因启动子59029位点SNPs与HIV-1感染者疾病进展关系的研究,按照纳入和排除标准选取文献,评价文献质量,提取相关数据。用Stata 10.0统计软件对数据进行Meta分析。结果按照纳入和排除标准,共纳入4个病例对照研究,其中长期不进展者（LTNP）总例数375人,快速进展者（RP）总例数282人。Meta分析结果显示,在排除了CCR5Δ32和CCR2-64I突变后,RP组的59029A/A纯合子基因型频率是0.235,高于LTNP组的0.144,两者差异具有统计学意义（Z=2.37,P=0.018）,OR及95%CI是3.441（1.236,9.582）。RP组的59029A等位频率是0.513,也高于LTNP组的0.370（Z=3.13,P=0.002）。结论 CCR5基因启动子59029A等位与HIV-1疾病的快速进展相关。     

HIV辅助受体CCR5基因多态性及其研究进展

辅助受体是HIV病毒感染人体所必须的，CCR5是HIV-1感染初期的主要辅助受体。本文主要讲述了CCR5的基因多态性，及基因多态性对HIV感染和AIDS病程进展的影响和作用原理，并讨论了CCR5与其它辅助受体和配体的相互作用，以及CCR5在治疗艾滋病药物开发中的应用前景。     

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.     

Differential pattern of CCR1 internalization in human eosinophils: prolonged internalization by CCL5 in contrast to CCL3

BACKGROUND: Whereas recent studies underlie the fundamental importance of the CC chemokine receptor 3 (CCR3) for the recruitment of eosinophils in allergic diseases, controversial data exist about the relevance of CCR1 on eosinophils. Therefore, the purpose of this study was to investigate the expression and regulation of CCR1 on eosinophils. METHODS: Flow cytometric analysis of whole blood eosinophils and CD16-negative selected eosinophils from healthy nonatopic donors and from patients with atopic disorders was performed and CCR1 receptor internalization and re-expression were studied. RESULTS: Flow cytometric analysis of whole blood eosinophils revealed that 17.8% of the donors expressed high levels of CCR1 (CCR1high) and 82.2% low levels of CCR1 (CCR1low). A significant down-regulation of CCR1 was induced by 24 h preincubation of isolated eosinophils from CCR1high donors either with IL-3, CC chemokine ligand 3 (CCL3), CCL5, CCL7, or CCL13. Internalization experiments using eosinophils from CCR1high donors revealed that CCL5 is more effective to induce CCR1 internalization than CCL3. Whereas CCR1 re-expression after stimulation with CCL3 reached prestimulation levels (120 min: 81.3% relative CCR1 surface expression) CCL5 induced a prolonged CCR1 internalization (120 min: 15.7%). CONCLUSIONS: This study demonstrates a distinct pattern of CCR1 internalization and re-expression in human eosinophils between CCL3 and CCL5, as CCL5 induces a prolonged CCR1 internalization and the basic value is not reached after 24 h. Since prolonged receptor internalization plays a central role in chemokine-mediated inhibition of receptor function, CCR1 seems to be an attractive target on human eosinophils for chemokine receptor blockade besides CCR3.     

Role of C-C chemokine receptors 1 and 5 and CCL3/macrophage inflammatory protein-1alpha in the cutaneous Arthus reaction: possible attenuation of their inhibitory effects by compensatory chemokine production

The deposition of immune complexes induces an acute inflammatory response with tissue injury. Immune complex-induced tissue injury is mediated by inflammatory cell infiltration that is highly regulated by multiple chemokines. To assess the role of the chemokine receptors CCR1 and CCR5, and a ligand for these receptors CCL3/macrophage inflammatory protein-1alpha, in this pathogenic process, the reverse passive cutaneous Arthus reaction was induced in mice lacking CCR1, CCR5, or CCL3. Edema was significantly attenuated in CCR1-deficient (CCR1(-/-)) and CCL3(-/-) mice but not CCR5(-/-) mice, compared with wild-type mice. Numbers of infiltrating neutrophils and mast cells were reduced in CCL3(-/-) and CCR1(-/-) mice, respectively, compared with wild-type mice. CCR1 and CCR5 were expressed on neutrophils and mast cells. Remarkably, the intradermal mRNA expression of CCL5/RANTES, another ligand for CCR1 and CCR5, was increased in CCR5(-/-) and CCL3(-/-) mice, compared with wild-type mice, while the cutaneous CCL3 mRNA expression was augmented in CCR1(-/-) and CCR5(-/-) mice. These results indicate that CCR1, CCR5, and CCL3 cooperatively contribute to the cutaneous Arthus reaction, and also suggest that enhanced expression of CCL3 and CCL5 compensates for the loss of CCR1, CCR5, and CCL3 in the reaction.     

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.     

CCR8-dependent activation of the RAS/MAPK pathway mediates anti-apoptotic activity of I-309/ CCL1 and vMIP-I

We have previously shown that the CC-chemokine 1-309 (CCL1) protects mouse thymic lymphomas against corticoid-induced apoptosis. Here, we analyzed the signal transduction pathways involved in this activity on BW5147 lymphoma. Inhibition of the CCL1 activity by pertussis toxin suggested the involvement of a G protein-coupled chemokine receptor. The role of CCR8 was supported by the observation that vMIP-I, another CCR8-ligand identified from the genome of a T cell transforming herpes virus, shared CCL1 anti-apoptotic activity. In addition to CCR8, BW5147 cells also expressed the CXCR4 receptor but its ligand, SDF-1 (CXCL12) showed only a modest anti-apoptotic activity. Other chemokines acting on CCR2, CCR4 and CCR5 failed to protect against apoptosis and to induce BW5147 chemotaxis, suggesting that these receptors were not functionally expressed. By contrast, both CCL1 and vMIP-I up-regulated ERK1/2 MAPK phosphorylation in BW5147 cells. Further analysis demonstrated that CCL1 activates the MAPK pathway in CCR8-transfected CHO cells. The implication of this pathway was confirmed by the fact that PD98059, an inhibitor of MEK kinases, as well as a dominant negative isoform of the M-RAS protein specifically blocked the anti-apoptotic activity of CCL1.     

Influence of nucleotide polymorphisms in the CCR2 gene and the CCR5 promoter on the expression of cell surface CCR5 and CXCR4

Polymorphisms in the CCR2 gene (CCR2-64I) and the CCR5 promoter (pCCR5-59029G) have been correlated with slower HIV-1 disease progression. How these polymorphisms influence the rate of AIDS progression has remained unclear. We have therefore investigated whether these nucleotide polymorphisms will reduce the expression levels of surface CCR5 and CXCR4, and thus lead to slower AIDS progression. For this, a cohort of Chinese volunteers in Taiwan was subjected to the determination of CCR2 and pCCR5 genotypes followed by analysis of the surface CCR5 and CXCR4 expression on five cell types derived from peripheral blood mononuclear cells by flow cytometry. Several significant associations were detected between genotypes and expression levels of the proteins. The most important finding was that an increased number of CD4(+) cells expressing CCR5 correlated with pCCR5-59029A homozygosity without the interference of both the CCR2-64 and the CCR5 delta 32 (deleted 32 bp) mutations (P: = 0.0453), which is consistent with the previous data on the association of the genotype to AIDS progression. Since different genetic polymorphisms co-exist in human beings, the rate of AIDS progression as well as the risk of rheumatoid arthritis may be governed by the interplay of the array of nucleotide changes and their affected proteins.     

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.     

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。     

CCL25 and CCR9 is a unique pathway that potentiates pannus formation by remodeling RA macrophages into mature osteoclasts

This study elucidates the mechanism of CCL25 and CCR9 in rheumatoid arthritis (RA). RA synovial fluid (SF) expresses elevated levels of CCL25 compared to OA SF and plasma from RA and normal. CCL25 was released into RA SF by fibroblasts (FLS) and macrophages (MΦs) stimulated with IL-1β and IL-6. CCR9 is also presented on IL-1β and IL-6 activated RA FLS and differentiated MΦs. Conversely, in RA PBMCs neither CCL25 nor CCR9 are impacted by 3-month longitudinal TNF inhibitor therapy. CCL25 amplifies RA FLS and monocyte infiltration via p38 and ERK phosphorylation. CCL25-stimulated RA FLS secrete potentiated levels of IL-8 which is disrupted by p38 and ERK inhibitors. CCL25 polarizes RA monocytes into nontraditional M1 MΦs that produce IL-8 and CCL2. Activation of p38 and ERK cascades are also responsible for the CCL25-induced M1 MΦ development. Unexpectedly, CCL25 was unable to polarize RA PBMCs into effector Th1/Th17 cells. Consistently, lymphokine like RANKL was uninvolved in CCL25-induced osteoclastogenesis; however, this manifestation was regulated by osteoclastic factors such as RANK, cathepsin K (CTSK), and TNF-α. In short, we reveal that CCL25/CCR9 manipulates RA FLS and MΦ migration and inflammatory phenotype in addition to osteoclast formation via p38 and ERK activation.     

CCL21 is sufficient to mediate DC migration,maturation and function in the absence of CCL19

Mice deficient in CCR7 signals show severe defects in lymphoid tissue architecture and immune response. These defects are due to impaired attraction of CCR7⁺ DC and CCR7⁺ T cells into the T zones of secondary lymphoid organs and altered DC maturation. It is currently unclear which CCR7 ligand mediates these processes in vivo as CCL19 and CCL21 show an overlapping expression pattern and blocking experiments have given contradictory results. In this study, we addressed this question using CCL19‐deficient mice expressing various levels of CCL21. Complete deficiency of CCL19 and CCL21 but not CCL19 alone was found to be associated with abnormal frequencies and localization of DC in naïve LN. Similarly, CCL19 was not required for DC migration from the skin, full DC maturation and efficient T‐cell priming. Our findings suggest that CCL21 is the critical CCR7 ligand regulating DC homeostasis and function in vivo with CCL19 being redundant for these processes.