Purvis S F, Jacobberger J W, Sramkoski R M, Patki A H, Lederman M M
Department of Environmental Health Sciences, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106, USA.
AIDS Res Hum Retroviruses. 1995 Apr;11(4):443-50. doi: 10.1089/aid.1995.11.443.
Jurkat cells stably expressing high levels of the HIV-1 Tat protein were generated after transfection with an Epstein-Barr virus-based episomal replicon and selection in hygromycin B. The Jurkat Tat transfectants exhibited a longer doubling time when compared to Jurkat cells or Jurkat cells transfected with the control parent plasmid. Cell cycle analysis revealed comparable durations of each phase of the cell cycle in the Tat and control transfectants. Flow cytometric analysis using Hoechst 33342 and propidium iodide staining revealed that the Tat transfectants exhibited a higher percentage of apoptotic cells when compared to the control transfectants (29.1 +/- 3.1 vs. 11.43 +/- 3.1%). Incubation of Jurkat cells with recombinant HIV-1 Tat protein resulted in induction of apoptosis. The HIV-1 Tat protein induces apoptosis in a CD4-positive T cell line. Tat-induced programmed cell death may contribute to the lymphocyte depletion seen in persons infected with HIV-1.
用基于爱泼斯坦-巴尔病毒的附加型复制子转染并在潮霉素B中筛选后，产生了稳定表达高水平HIV-1 Tat蛋白的Jurkat细胞。与Jurkat细胞或用对照亲本质粒转染的Jurkat细胞相比，Jurkat Tat转染子表现出更长的倍增时间。细胞周期分析显示，Tat转染子和对照转染子的细胞周期各阶段持续时间相当。使用Hoechst 33342和碘化丙啶染色的流式细胞术分析显示，与对照转染子相比，Tat转染子表现出更高比例的凋亡细胞(29.1±3.1%对11.43±3.1%)。用重组HIV-1 Tat蛋白孵育Jurkat细胞导致凋亡诱导。HIV-1 Tat蛋白在CD4阳性T细胞系中诱导凋亡。Tat诱导的程序性细胞死亡可能导致HIV-1感染者出现淋巴细胞耗竭。
