中国汉族人HIV辅助受体CCR5编码区新SNP位点研究

目的：普查中国汉族人群HIV－1协同受体CCR5编码区的基因多态性位点，为中国的艾滋病防治提供依据。方法：CCR5编码区用2对引物进行PCR扩增，设计测序引物依次测序，样本数为45例，用DNAstar分析测序结果，寻找SNP位点。结果：在编码区共发现6个SNP位点，4个引起氨基酸改变：A184G、G503T、G668A、G999T；一个单碱基缺失，引起移码突变和提前终止。A184G、G503T、G999T，三个中国汉族人所特有的SNP位点为首次发现，等位基因频率分别为1％，39．5％和9．5％；其中G503T分布明显不符合Hardy-Weinberg平衡。G668A和894C缺失曾在中国和日本人群中发现过，但我们的结果与所报道的基因频率不完全一致。结论：中国汉族人CCR5编码区SNP位点有自己的特点，与高加索人和非洲人明显不同，与日本人也不完全一致。我们共找到5个引起氨基酸改变的突变或SNP位点，其中3个为首次发现，这些SNP对于HIV－1感染和艾滋病病程的影响值得进一步研究。     

CCR5基因编码区894C缺失突变在中国人群中的发现

目的：调查中国汉族人HIV协同受体CCR5编码区基因突变和SNP特点。方法：PCR扩增CCR5编码区，PCR产物直接进行基因测定。结果：在50例标本中，发现一例CCR5X编码区894C杂合子；采用反向经物进行验证，确定测定结果准确无误，该人缺失引起移码突变，使CCR5C端减少了44个氨基酸，结论：中国汉族人群中存在CCD5 894C，该突变能导致CCR5受体结构和功能改变。     

CCR5基因编码区894C缺失突变在中国人群中的发现

目的 调查中国汉族人HIV协同受体CCR5编码区基因突变和SNP特点。方法PCR扩增CCR5编码区,PCR产物直接进行基因测定。结果 在50例标本中,发现一例CCR5X编码区894ΔC杂合子;采用反向经物进行验证,确定测序结果准确无误。该人缺失引起移码突变,使CCR5C端减少了44个氨基酸。结论 中国汉族人群中存在CCR5 894ΔC,该突变能导致CCR5受体结构和功能改变。     

中国艾滋病人群中HIV—1抗性基因多态性及其与病程发展关系分析

目的：分析了解中国艾滋病病人中CCR－5,CCR－2b和SDF－1基因型组成和等位基因突变频率及其与病程发展的关系。方法：应用聚合酶链反应（PCR）方法对68份艾滋病病人外周血单核细胞样本进行扩增,获得人基因组中CCR－5,CCR－2b和SDF－1基因片段,通过限制性片段长度多态性和／或DNA测序分析CCR5－Δ32,CCR4－m303,CCR2b-64I和SDF1－3’A的等位基因突变频率。结果：68例艾滋病病人中未发现有CCR5－Δ32和CCE5m303基因突变,CCR2b-64I和SDF1－3’A等位基因突变频率分别为21．32％和27．94％,95％CI 11．6％－31．0％和17．2％－38．6％CCR2b-64I和SDF1－3’A等位突变的群体分布符合Hardy-Weinberg规律。SDF1－3’A／CCR2b-64I,SDF1-3’A,CCR2b-64I等位基因突变型和野生型个体发展为AIDS的潜伏期分别为7年,5．8年,5．2年和4．6年。结论：目前发现的HIV感染者均为无CCR5－Δ32和CCR5－m303的易感人群,本研究首次阐明了中国HIV感染者人群中CCR5－Δ32,CCR5－m303,CCR2b-64I和SDF1－3’A等位基因突变频率和多态性特点,发现CCR2b-64I和SDF1－3’A等位基因突变可延缓艾滋病病程的发展,为深入研究HIV－1抗性基因对中国人群HIV－1感染的遗传易感性及其在艾滋病病程发展中的作用具有指导性意义。     

1型糖尿病患者及其一级亲属趋化因子受体CCR2和CCR5的基因多态性研究

目的　探讨趋化因子受体CCR2、CCR5基因多态性与 1型糖尿病 (T1DM)及其一级亲属之间的关系 ,并与T2DM及非DM对照进行比较。　方法　利用PCR方法检测 48例T1DM及其 5 7名一级亲属CCR5的基因多态性 ,利用BsaBI限制性内切酶的PCR RFLP方法检测T1DM及其一级亲属的CCR2基因多态性。　结果　T1DM组、T1DM一级亲属组、T2DM组分别与非DM对照组比较 ,CCR5Δ32突变的等位基因频率和基因型频率差异无显著意义。T1DM组CCR2 6 4I突变基因型及等位基因频率均高于非DM对照组 (P <0 .0 1) ;T2DM组CCR2 6 4I突变基因型及等位基因频率均低于T1DM组 (P <0 .0 1)。　结论　CCR5Δ32突变与T1DM无显著相关性。CCR2 6 4I突变与T1DM发病显著相关 ,CCR2基因可能是T1DM一个新的候选基因。     

广西艾滋病疫情严重地区壮族和汉族普通人群CCR5 CCR2 SDF1等位基因检测及其意义

目的调查艾滋病病毒Ⅰ型（HIV—1）感染相关的CCR5、CCR2及SDF1等位基因,在广西壮族和汉族普通人群中的多态性及其流行病学意义。方法在广西天等县和南宁市,选取150名壮族和90名汉族健康人作为研究对象,采集外周血,提取基因组DNA,用聚合酶链反应（PCR）扩增CCR5、CCR2及SDF1基因的特定片段。直接根据PCR扩增结果分析CCR5基因多态性,运用限制性片段长度多态性（RFLP）技术分析CCR2及SDF1的多态性。结果全部研究对象的CCR5基因均为野生型,未发现CCR5△32突变;CCR2—64Ⅰ等位基因频率在壮、汉族普通人群中分别为25．7％、26．1％;SDF1—3’A等位基因频率分别为27．7％和27．2％。结论广西壮、汉族普通人群缺乏艾滋病抗性的CCR5△32等位基因,而CCR2-641和SDF1—3’A等位基因频率较高。该研究为深入研究广西壮、汉族普通人群对于HIV—1感染的遗传易感性,以及对艾滋病疫情和病程的影响提供了比较全面、可靠的数据。     

先天性长QT综合征KVLQT1和HERG基因新突变位点的检测

目的：研究中国人先天性长QT综合征（long QT syndrome,LQTS)HERG和KVLQT1的基因突变情况。方法,利用聚合酶链反应和DNA测序对11个LQTS家系HERG跨膜编码区S1－S6和KVLQT1跨膜编码区S2－S6进行基因检测。结果（1）11个LQTS患者在国外已知突变点均无突变;（2）发现4个新的错义突变位点,分别为T1515G（HERG）,C682T,C934T,G983A（KVLQT1）。其对应的氨基酸改变为E505D,R228C,S230L,P312S和R328C。结论：在中国人LQTS患者HERG和KVLQT1上发现了4个新的基因突变位点。     

广西壮族人Caveolin-3基因多态性与2型糖尿病的关系

目的探讨广西壮族人Caveolin-3基因外显子单核苷酸多态性（SNP）与2型糖尿病（T2DM）的关系。方法选择24例T2DM（T2DM组）患者及10例正常对照者（对照组）,应用PCR法对两组广西壮族人Caveolin-3基因外显子SNP进行PCR扩增后测序。结果 Caveolin-3基因第2外显子非编码区有2个位点发生突变,分别是12842 A→G和12715 A→T。2位点的等位基因频率在糖尿病组和正常对照组存在统计学差异（P〈0.05）。12842A→G和12715 A→T位点有三种联合基因型（AT、GT、GA）,其中AT、GA型的分布频率在两组中比较P〈0.05。结论 Caveolin-3基因12842 A→G和12715 A→T位点突变可能与T2DM的发病有关。     

蛋白酪氨酸磷酸酶1B基因多态性与2型糖尿病和肥胖的相关性研究

目的 研究中国人群中蛋白酪氨酸磷酸酶1B（PTP-1B）基因的单核苷酸多态性（SNP）与2型糖尿病及肥胖的相关性。方法 采用直接测序法对PTP—1B基因作SNP筛查，并在夫妻配对样本中对所检出的SNP作基因分型。结果 共检出6个SNPs位点，其中内含子区3个（15／37C→A，16／82A→G，17／301C→T），外显子区3个（E8／45C→T，E9／35G→A，E10／372G→A），其中E9／35G→A为新发现的突变类型；在病例-配偶对照研究中发现，15／37C→A，16／82A→G和17／301C→T等位基因频率在糖尿病患者和正常人配偶中差异有统计学意义（均P〈0．05），其余位点的等位基因频率在两组间的分布则无明显差异。与肥胖的相关性研究中发现15／37C→A和17／301C→T位点与男性的腰臀比（WHR）相关（P〈0．05）。结论 PTP-1B基因的SNP位点15／37C→A，16／82A→G和17／301C→T多态性可能和2型糖尿病的发病相关，其中15／37C→A和17／301C→T与男性的WHR相关。     

深圳地区艾滋病病毒感染人群的HIV—1抗性基因多态性研究

目的 研究HIV－1相关的等位基因CCR5△32、CCR5m303、CCR2b和SDF1在艾滋病病毒感染人群中的分布,为深入分析这些遗传突变在HIV－1感染、艾滋病发病进程中的作用提供理论依据。方法 收集深圳地区近年来艾滋病毒感染者的血液标本共91份,提取基因组DNA。经PCR或PCR－ RFLP分析,计算遗传突变频率、纯合子和杂合子的构成比例。采用x^2检验或t检验进行群体分布的拟合度和差异的显著性分析。结果 在所有被检测的91例HIV－1感染者的个体中,未发现有CCR5△32和CCR5m303的遗传突变,CCR2b－641和SDF1－3’A的突变频率较高,分别为21．43％和25．82％。与中国普通汉族人的结果相近。结论 研究结果提示深圳地区的艾滋病病毒感染者本身对HIV－1病原体可能具有较大的遗传易感性。由于他们具有较高的CCR2b-641和SDF1－3’A突变频率,后者在艾滋病发病过程中的影响值得进一步研究。     

Heterozygosity for CCR5-Delta32 but not CCR2b-64I protects against certain intracellular pathogens

Objective

To determine the association between CCR5 and CCR2b genotype and the clinical manifestation of first and subsequent AIDS‐defining illnesses (ADIs).

Methods

The distribution of ADIs was examined by CCR5 and CCR2b genotype in a subset of homosexual men enrolled in the Sydney AIDS Prospective Study. The expected number of ADIs was calculated from rates observed in the same tertiary hospital over the same period.

Results

Information on initial ADI was collected for 117 homosexual men diagnosed with AIDS before January 1998. Of these individuals, 17 were heterozygous for the CCR5‐DΔ32 mutation and 11 were heterozygous for CCR2b‐64I. The number of observed cases of Pneumocystis carinii pneumonia (PCP), toxoplasmosis, Mycobacterium avium complex (MAC) and cryptosporidiosis reported as a first ADI was substantially fewer in people heterozygous for the CCR5‐DΔ32 mutation than for those without the mutation, despite similar age, CD4 T‐cell count at AIDS diagnosis, year of AIDS diagnosis and receipt of antiretroviral treatment. In addition, among individuals heterozygous for CCR5‐DΔ32 there were fewer cases of PCP, toxoplasmosis, MAC, and cryptosporidiosis observed as subsequent ADIs compared to the number expected, based on rates measured in the same hospital during the same period (seven observed vs. 24 expected, RR = 0.3, 95% CI = 0.01–0.6). The distribution of first and subsequent ADIs did not differ from the number expected in individuals heterozygous for the CCR2b‐64I mutation.

Conclusion

Results from this study show that heterozygosity for CCR5‐DΔ32 but not CCR2b‐64I appears to protect against opportunistic infections.

     

CCR2在结直肠癌的表达及意义

目的探讨大肠癌中CCR2蛋白的表达与肿瘤病理特征的关系。方法用免疫组织化学方法检测148例大肠癌患者手术切除大肠癌组织及66例癌旁对照组织中CCR2表达情况。结果大肠癌组织中CCR2的表达明显高于大肠正常组织（P〈0．01）,并且其表达强度与DukeC＋D期、肿瘤淋巴结转移、远处脏器转移以及较低的分化程度有关（各组中P〈0．01,差异具有统计学意义）;与年龄、性别、肿瘤发生部位以及肿瘤大小无关。结论CCR2的过表达在大肠癌发生、发展及转移的过程中可能发挥一定作用。CCR2可以作为大肠癌分化程度较低且伴淋巴结转移、肝转移的预测指标之一。     

Possible contribution of chemokine receptor CCR2 and CCR5 polymorphisms in the pathogenesis of chronic spontaneous autoreactive urticaria

BackgroundAutoimmune mechanisms play a role in the pathophysiology of chronic urticaria. As the genetic background of autoimmunity is well proven, the role of genetics in chronic urticaria is hypothesised.Methods153 unrelated chronic spontaneous urticaria patients with a positive result of autologous serum skin test were included into the study, as were 115 healthy volunteers as control group. In all subjects we analysed CCR2 G190A and CCR5 d32 polymorphisms.ResultsWe noticed higher prevalence of CCR2 A allele as well as lower frequency of CCR5 d32 in chronic urticaria group in comparison to control group, with borderline statistical significance. Additionally, we assumed haplotype Gd statistically significant negative chronic urticaria association with tendency to higher frequency of Aw haplotype in this group.ConclusionsThe results of our study imply the role of autoimmune components in chronic urticaria pathogenesis and present chronic urticaria as possibly genetically related disorder.     

中国汉族人群中艾滋病相关的CCR2-64I和CCR5-C927T等位基因的多态性分析

目的 调查中国汉族人群中艾滋病相关的CCR2-64I和CCR5-C927T等位基因突变频率和多态性特点。方法 用试剂盒提取中国889例汉族人外周血单个核细胞的基因组DNA,随之进行聚合酶链反应/限制性片段长度多态性(PCR/RFLP)技术和DNA直接测序法分析,并对有关的数据进行统计学分析。结果 在检测的889例个体中,野生型CCR2-64I等位基因纯合子(基因型64V/64V)有570例,杂合子(64V/64I)298例,突变纯合子(64I/64I)21例,所占百分比分别是64.1％、33.5％和2.36％,CCR2-64I等位基因突变频率为0.1913。在889份基因样品中,有93例经过CCR5-C927T基因突变检测,结果显示野生型CCR5-C927T纯合子(基因型C/C)为63例,占67.7％;杂合子(C/T)为26例,占28.0％;突变纯合子基因型(T/T)为4例,占4.3％;CCR5-C927T等位基因频率为0.183。统计分析表明在我国汉族人群中,上述两种等位基因多态性呈Hardy-Weinberg平衡分布。结论 本研究首次阐明了土生土长的中国汉族人群中CCR2-64I和CCR5-C927T等位基因突变频率和多态性特点,这一结果将有助于综合评估中国人群对HIV-1感染的遗传易感性,同时为深入研究HIV-1抗性基因在艾滋病发病机制中的作用奠定了基础。     

CCR1 knockdown suppresses human non-small cell lung cancer cell invasion

Purpose  CC chemokine receptor 1 (CCR1) plays a critical role in the recruitment of leukocytes to the site of inflammation. Tumor invasion and metastasis share many similarities with leukocyte trafficking, which is critically regulated by chemokines and their receptors. In this study, we aimed to assess the role of CCR1 in non-small cell lung cancer (NSCLC). Methods  CCR1 expression was determined by Western blotting in two human NSCLC clones (95C and 95D) with different metastatic potential. We silenced CCR1 expression through microRNA-mediated RNA interference, and examined the invasiveness and proliferation of CCR1-silenced NSCLC cell through Matrigel assay and MTT assay. Matrix metalloproteinases (MMPs) activity was determined by gelatin zymography. Results  We found that expression of CCR1 was correlated with the aggressive phenotype of the NSCLC cells. CCR1 knockdown significantly suppressed the invasiveness of NSCLC cells, but had only a minor effect on cell proliferation. Moreover, we demonstrated that CCR1 knockdown significantly reduced the expression level of matrix metalloproteinase-9. Conclusions  These findings suggest that CCR1 contributes to NSCLC cell migration by stimulating cell invasion, independent of cell proliferation. CCR1 might be a new target for NSCLC therapy. C.-L. Wang and B.-S. Sun contributed equally to this work.     

Evolution of CCR5 and CCR2 Genes in Bats Showed Multiple Independent Gene Conversion Events

Chemokine receptors are an important determinant for the infectiousness of different pathogens, which are able to target the host cells by binding to the extracellular domains of these proteins. This is the mechanism of infection of HIV-1, among other concerning human diseases. Over the past years, it has been shown that two chemokine receptors, CCR2 and CCR5, have been shaped by events of gene conversion in different mammalian lineages, which has been linked to a possible selective advantage against pathogens. Here, by taking advantage of available bat genomes, we present the first insight of CCR2 and CCR5 evolution within the Chiroptera order. In total, four independent events of recombination between CCR2 and CCR5 were detected: two in a single species, Miniopterus natalensis; one in two species from the Rhinolophoidea superfamily; and one in four species from the Pteropodidae family. The regions affected by the gene conversions were generally extensive and always encompassed extracellular domains. Overall, we demonstrate that CCR2 and CCR5 have been subject to extensive gene conversion in multiple species of bats. Considering that bats are known to be large reservoirs of virus in nature, these results might indicate that chimeric CCR2-CCR5 genes might grant some bat species a selective advantage against viruses that rely in the extracellular portions of either CCR2 or CCR5 as gateways into the cell.     

CCR5: From Natural Resistance to a New Anti-HIV Strategy

The C-C chemokine receptor type 5 (CCR5) is a key player in HIV infection due to its major involvement in the infection process. Investigations into the role of the CCR5 coreceptor first focused on its binding to the virus and the molecular mechanisms leading to the entry and spread of HIV. The identification of naturally occurring CCR5 mutations has allowed scientists to address the CCR5 molecule as a promising target to prevent or limit HIV infection in vivo. Naturally occurring CCR5-specific antibodies have been found in exposed but uninfected people, and in a subset of HIV seropositive people who show long-term control of the infection. This suggests that natural autoimmunity to the CCR5 coreceptor exists and may play a role in HIV control. Such natural immunity has prompted strategies aimed at achieving anti-HIV humoral responses through CCR5 targeting, which will be described here.