Zhang L, Huang Y, Yuan H, Chen B K, Ip J, Ho D D
The Aaron Diamond AIDS Research Center, The Rockefeller University, New York, New York 10016, USA.
J Virol. 1997 Jul;71(7):5608-13. doi: 10.1128/JVI.71.7.5608-5613.1997.
Human immunodeficiency virus type 1 (HIV-1)-infected individuals who remain asymptomatic despite prolonged infection present a unique opportunity to understand virologic and immunologic factors involved in the pathogenesis of AIDS. We have previously identified a group of long-term survivors (LTS) who are clinically healthy and immunologically normal despite 13 to 15 years of HIV-1 infection. In this study, we examined the 5' long terminal repeat (5' LTR) sequences in eight of these LTS. A total of 29 nucleotide sequences were obtained from their peripheral blood mononuclear cells (PBMC). Analysis of these sequences revealed no gross deletions within the 5' LTR. Seven of the eight subjects shared nearly identical consensus sequences in the binding sites for NF-kappaB, Sp1, and the viral trans-activator Tat. In multiple samples from one individual (Pt 5), however, G-to-A hypermutations were found throughout the entire region, suggesting a genetically defective 5' LTR. The effects of the observed genetic variations on LTR transcription were studied by transient transfection of an LTR-driven luciferase reporter gene and by infection with a full-length recombinant HIV-1 containing a luciferase reporter (HIVHXBLTRluc). A wide range of basal and Tat-induced transcriptional activities was found among the 5' LTR from seven of the eight LTS in both transfected 293 cells and donor PBMC, suggesting a functionally intact 5' LTR in these individuals. It is therefore unlikely that defects in the 5' LTR are the underlying explanation for the benign clinical course associated with these seven individuals. However, functional abnormalities were found in the LTR from Pt 5 in directing both heterologous and viral gene expression, providing a possible genetic explanation for the low viral load and prolonged asymptomatic state of this individual. Last, a similar overall degree of genetic diversity was found among viruses from the LTS compared to those from patients with AIDS, reinforcing the notion that a strong correlation between the degree of genetic diversity and the rate of disease progression is unlikely.
1型人类免疫缺陷病毒(HIV-1)感染个体尽管感染时间延长但仍无症状,这为了解艾滋病发病机制中涉及的病毒学和免疫学因素提供了独特机会。我们之前鉴定出一组长期存活者(LTS),他们尽管感染HIV-1达13至15年,但临床健康且免疫功能正常。在本研究中,我们检测了其中8名LTS的5'长末端重复序列(5' LTR)。从他们的外周血单核细胞(PBMC)中总共获得了29个核苷酸序列。对这些序列的分析显示5' LTR内没有明显缺失。8名受试者中有7名在核因子κB(NF-κB)、Sp1和病毒反式激活因子Tat的结合位点共享几乎相同的共有序列。然而,在一名个体(患者5)的多个样本中,整个区域都发现了G到A的超突变,提示5' LTR存在基因缺陷。通过瞬时转染由LTR驱动的荧光素酶报告基因以及用含有荧光素酶报告基因的全长重组HIV-1(HIVHXBLTRluc)感染,研究了观察到的基因变异对LTR转录的影响。在转染的293细胞和供体PBMC中,8名LTS中的7名的5' LTR表现出广泛的基础和Tat诱导的转录活性,表明这些个体的5' LTR功能完整。因此,5' LTR缺陷不太可能是这7名个体临床病程良性的根本原因。然而,在患者5的LTR中发现了在指导异源和病毒基因表达方面的功能异常,这为该个体病毒载量低和无症状状态延长提供了可能的遗传学解释。最后,与艾滋病患者的病毒相比,LTS的病毒之间发现了相似的总体遗传多样性程度,这强化了遗传多样性程度与疾病进展速度之间不太可能存在强相关性的观点。