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作 者:任国良[1] 陈健平[1] 贾明明[1] 寇中琛 刘沙[1] 马鹏飞[1] 邵一鸣 洪坤学[1]
机构地区:[1]中国疾病预防控制中心性病艾滋病预防控制中心传染病预防控制国家重点实验室,北京100050 [2]Biomed Immunotech, University of Florida, Gamesville, FL33612
出 处:《中华预防医学杂志》2009年第5期404-408,共5页Chinese Journal of Preventive Medicine
基 金:国家重点基础研究发展计划(2006CB504207)
摘 要:目的研究HIV-1感染者CD8^+T细胞受体(TCR)基因的多样性变化特征及其与病毒载量的相关性。方法应用抗CD8单克隆抗体从9份HIV-1感染者和7份HIV-1阴性对照样本外周血单个核细胞中分离CD8^+T细胞,提取总RNA,然后采用一步(one step)及巢式(nested)多聚酶链式反应(PCR)的方法对22个T细胞受体VB基因家族的互补决定区3(CDR3)进行扩增,利用ABI-3700测序仪对扩增的PCR产物进行扫描,定量分析HIV-1感染者TCRCDR3区多样性变化特征及其与病毒载量的相关性。结果HIV-1感染者和正常人相比较其CD8^+T细胞的TCR基因多样性明显减少,部分TCR Vβ基因家族CDR3区的高斯分布破坏;TCR的紊乱与病毒载量呈正相关(r=0.771,P〈0.05);HIV-1感染引起患者TCR多样性的改变不仅表现在不同Vβ基因家族上,而且也表现在CDR3长度上,其中感染者Vβ2、Vβ4、Vβ5、Vβ17、Vβ20、Vβ21、Vβ23及Vβ24基因家族的变化与正常人存在统计学差异。结论HIV-1感染能引起CD8^+T细胞TCR基因多样性的减少及高斯分布的破坏,TCR CDR3区的紊乱与病毒载量呈正相关。Objective To determine the complementary determining region 3 (CDR3) length diversity of T cell receptor Vβ repertoires of CD8^+ T lymphocytes and to explore its association with viral load in individuals with HIV-1 infection. Methods Separation of CD8 + T cells from peripheral blood mononuclear cells (PBMCs) was carried out by using immunomagnetic beads coated with anti-CD8 antibody. Total RNAs from the purified CD8^+ T lymphocytes were isolated and used to perform polymerase chain reaction (PCR) amplifications in CDR3 of 22 T cell receptor (TCR) gene families. CDR3 diversity and its association with viral load in individuals with HIV-1 infection were analyzed. Results An average diversity for all CDR3 profiles in CD8^+ T cells from 9 HIV-infected individuals was significantly different as compared to 7 age-matched healthy donors (P 〈 0. 05) with the HIV-infected individuals losing diversity in the CDR3 profiles. There was positive correlation between changes in TCR CDR3 diversity and viral load (r =0. 771 ,P〈0. 05). The changes in CDR3 length diversity of Vβ families in HIV-infected individuals, particular in Vβ2 ,Vβ4, Vβ5, Vβ17, Vβ20, Vβ21, Vβ23, Vβ24, were statistically different from the healthy controls. Conclusion HIV-1 infection might induce the loss of TCR Vβ repertoire diversity and disrupt the CDR3 distributions within CD8^+ Y cells. There should be positive correlation between changes in TCR CDR3 diversity and the viral load in HIV-1 infected patients.
关 键 词:CD8阳性T淋巴细胞 HIV-1 受体 抗原 T细胞
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