首都医科大学学报 ›› 2011, Vol. 32 ›› Issue (4): 449-452.doi: 10.3969/j.issn.1006-7795.2011.04.002

• 皮肤病与性病学专题 • 上一篇    下一篇

Alu-LTR PCR方法检测HAART治疗系列各细胞亚群的整合型HIV-1 DNA

李娟, 焦艳梅, 高艳青, 寇卜心, 张彤, 吴昊   

  1. 首都医科大学附属北京佑安医院性病艾滋病临床诊疗中心,北京 100069
  • 收稿日期:2011-02-25 修回日期:1900-01-01 出版日期:2011-08-21 发布日期:2011-08-21

Optimization of Alu-LTR PCR approach to detect integrated HIV-1 DNA in variety of cell subsets at different stages after HAART

LI Juan, JIAO Yan-mei, GAO Yan-qing, KOU Bu-xin, ZHANG Tong, WU Hao   

  1. Clinical Center of STDS and AIDS, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China
  • Received:2011-02-25 Revised:1900-01-01 Online:2011-08-21 Published:2011-08-21

摘要: 目的 应用Alu-LTR PCR方法对高效抗反转录病毒治疗(highly active antiretroviral therapy,HAART)系列的CD4+T,CD8+T及B细胞进行检测,明确不同细胞亚群及HAART治疗的不同阶段是否存在整合的HIV-1 DNA。方法 收集20例HIV-1感染患者HAART治疗过程中0、4、12周及10例健康对照的抗凝血标本,纯化CD4+T,CD8+T及B细胞并提取DNA,应用Alu-LTR PCR方法进行检测。结果 20例HIV-1感染者HAART治疗系列中CD4+T细胞及CD8+T细胞成功扩出目的片段,CD8+T细胞所扩条带亮度均低于CD4+T细胞,HAART治疗系列0、4、12周标本所扩条带亮度没有明显差异。B细胞及10例健康者均为阴性。结论 CD4+T及CD8+T细胞存在整合型HIV-1 DNA,HIV储藏库主要存在于CD4+T细胞内。B细胞内不存在整合型HIV-1 DNA。随着HAART治疗的进程,整合型HIV-1 DNA仍然存在,进一步证明HAART不能根除HIV储藏库。

关键词: Alu-LTR PCR, 整合型人类免疫缺陷病毒-1, CD4+T细胞, CD8+T细胞, B细胞

Abstract: Objective To determine if there is integrated human immunodeficiency virus-1(HIV-1) DNA in variety of cell subsets at different stages after highly active antiretroviral therapy(HAART) through detecting HIV-1 DNA in CD4+T cell, CD8+T cell and B cell by Alu-long terminal repeat(LTR) polymerase chain reaction(PCR). Methods The genomic DNA from purified CD4+T, CD8+T, B cells of 20 HIV-1 patients after HAART 0, 4, 12 W and 10 healthy control subjects was obtained by Alu-LTR PCR. Results After Alu-LTR PCR, integrated HIV-1 was detected in each of the 20 cases of HIV/AIDS of CD4+T cells and CD8+T cells after HAART. The product of CD8+T cells were lower than that of the CD4+T cells. The product of HAART 4, 0, 12 W had no significant difference. Conclusion There exists integrated HIV-1 DNA in CD4+T cells and CD8+T cells, and HIV viral reservoir mainly exists in CD4+T cells. Integrated HIV-1 DNA do not exist in B cells. Integrated HIV-1 DNA still exist, and HAART do not eradicate HIV viral reservoir.

Key words: Alu-LTR PCR, integrated human immunodeficiency virus-1, CD4+T cell, CD8+T cell, B cell

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