艾滋病患者抗病毒治疗初期不同部位HIV-1DNA比较研究

Comparison of HIV-1 DNA in different parts of early antiretroviral treatment patients

目的了解抗逆转录病毒治疗早期，外周血、肠黏膜相关淋巴组织及淋巴结中单个核细胞中HIV-1DNA量的差异。方法收集2006至2011年首都医科大学附属北京佑安医院规律随诊的留取有外周血、肠黏膜相关淋巴组织及淋巴结标本的HIV-1／AIDS患者，共11例。平均年龄39（25～55）岁，收集治疗前及治疗12周后患者的外周血、肠黏膜相关淋巴组织及淋巴结标本，用密度梯度离心法分离单个核细胞，用DNA抽提试剂盒提取DNA，用实时定量聚合酶链反应的方法检测HIV-1DNA拷贝数。用非参数检验方法分析比较各实验组之间HIV-1DNA拷贝数的差异。结果治疗前，不同部位HIV-1DNA量的差异具有统计学意义（χ2=17．636，P〈0．01），肠黏膜相关淋巴组织的HIV-1DNA量[（10714±2043）拷贝／10^6细胞]和淋巴结内[（9145±1202）拷贝／10^6细胞]高于外周血[（66±8）拷贝／10^6细胞，差异有统计学意义（U值均为0．00，P值均〈0．05）]，治疗前淋巴结与肠黏膜相关淋巴组织中的HIV-1DNA量之间差异无统计学意义（U=46．00，P〉0．05）；抗逆转录病毒治疗12周后，不同部位HIV一1DNA量的差异有统计学意义（χ2=22．000，P〈0．01）；肠黏膜相关淋巴组织的HIV-1DNA量[（1701±790）拷贝／10^6细胞]和淋巴结内[（11591±1781）拷贝／10^6细胞]高于外周血[（18±3）拷贝／10。细胞（U值均为0．00，P值均〈0．05）]；肠黏膜相关淋巴组织HIV-1DNA水平从平均10714拷贝／10^6细胞降低至平均1701拷贝／10^6细胞，差异有统计学意义（Z=-2．934，P〈0．05）；外周血HIV-1DNA水平从平均66拷贝／10。细胞降低至平均18拷贝／10^6细胞，差异有统计学意义（Z=-2．934，P〈0．05）。结论肠黏膜相关淋巴组织及淋巴结是HIV-1DNA储存的重要场所。

Objective To study HIV-1 DNA levels in different parts of HIV patients during the early stage of antiretroviral therapy. Methods The peripheral blood, gut associated lymphoid tissues and lymph nodes samples were collected before and 12 weeks after treatment in regular follow-up HIV-1/AIDS patients in Beijing Youan Hospital （ n = 11 ）. The average age was 39 years old （ 25 to 55）. Mononuelear Cells were isolated by density gradient centrifugation and then used DNA extraction kit to extract DNA. Real- time quantitative polymerase chain reaction was used to examine HIV-1 DNA copy-number. Non-parametric test was used to analyse the differences of HIV-1 DNA copy numbers among groups. Results Before treatment, HIV-1 DNA copy-number in both gut associated lymphoid tissues （10 714 ± 2043 ） copies/10^6 cells and lymph nodes （9145 ± 1202） copies/106 cells were higher than that in the peripheral blood （66 ± 8 ） copies/106 cells （ U = 0. 00 ,P 〈 0. 05 ） , There was no significant difference between lymph nodes and gut associated lymphoid tissues （ U = 46. 00,P 〉 0. 05 ）. After 12 weeks of treatment, HIV-1 DNA copy-number in both gut associated lymphoid tissues （1701 ± 790） copies/106 cells and lymph node （11 591 ± 1781 ） copies/106 cells were higher than the peripheral blood （ 18 ± 3） copies/106 cells （Z = - 2. 934 ,P 〈 0. 05）. There was a significant reduction of DNA copy-number in gut associated lymphoid tissues and peripheral blood after treatment （Z = - 2. 934,P 〈 0. 05）. Conclusion Gut associated lymphoid tissues and lymph nodes may be important latent reservoirs for HIV-1 DNA.