DETECTION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PROVIRUS IN MONONUCLEAR-CELLS BY INSITU POLYMERASE CHAIN-REACTION
DETECTION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PROVIRUS IN MONONUCLEAR-CELLS BY INSITU POLYMERASE CHAIN-REACTION
复制标题
DOI：
10.1056/nejm199205213262103
复制
发表时间：
1992-05-21
影响因子：
158.5
通讯作者：
POMERANTZ, RJ
中科院分区：
文献类型：
--
作者：
BAGASRA, O;HAUPTMAN, SP;POMERANTZ, RJ
背景人类免疫缺陷病毒1型（HIV-1）感染的研究试图定量病毒载量，并将其与免疫缺陷的程度相关联。在一项研究中，只有约1/10，000的外周血单核细胞（PBMC）表达HIV-1，但在其他研究中，至少有1/100的CD 4阳性细胞被感染并携带HIV-1前病毒。我们开发了一种新的，高灵敏度的原位聚合酶链反应（PCR）方法，扩增完整的单细胞内选定的遗传区域。我们应用该技术检测了不同疾病阶段感染者PBMC中携带HIV-1前病毒的比例。11例HIV-1阴性患者的PBMC经原位PCR检测均未检出HIV-1前病毒。在56例HIV-1感染者中，PBMC携带HIV-1的百分比范围为0.1%至13.5%。13例持续性全身淋巴结病患者（平均6.6%）和19例获得性免疫缺陷综合征（IV-A至IV-C期）患者（4.6%）中感染单核细胞的平均百分比高于19例无症状HIV-1感染患者（0.9%）（P < 0.001）。然而，在5例卡波西肉瘤（IV-D期）患者中，平均只有1.6%的单核细胞被感染。在HIV-1感染中，受感染的PBMC的比例似乎比先前描述的高至少10倍。很可能大多数感染细胞含有潜伏或缺陷形式的HIV-1前病毒，这在一些早期研究中没有检测到。
Background. Studies of human immunodeficiency virus type 1 (HIV-1) infection have attempted to quantitate the viral load and correlate it with the degree of immune deficiency. In one study, only about 1 in 10,000 peripheral-blood mononuclear cells (PBMC) expressed HIV-1, but in other studies, at least 1 in 100 CD4-positive cells was infected and harbored the HIV-1 provirus.Methods. We developed a new, highly sensitive in situ polymerase-chain-reaction (PCR) method that amplifies selected genetic regions within intact single cells. We used this technique to determine the proportion of PBMC carrying HIV-1 provirus in infected patients in different stages of disease.Results. None of the PBMC from 11 HIV-1-seronegative patients were found to be positive for HIV-1 provirus by the in situ PCR method. In 56 patients infected with HIV-1, the percentage of PBMC with HIV-1 ranged from 0.1 percent to 13.5 percent. The mean percentage of infected mononuclear cells was greater in 13 patients with persistent generalized adenopathy (mean, 6.6 percent) and 19 with the acquired immunodeficiency syndrome (Stages IV-A to IV-C) (4.6 percent) than in 19 patients with asymptomatic HIV-1 infection (0.9 percent) (P < 0.001). However, in five patients with Kaposi's sarcoma (Stage IV-D), an average of only 1.6 percent of mononuclear cells were infected Conclusions. In HIV-1 infection, the proportion of PBMC that are infected appears to be at least 10 times higher than previously described. It is likely that most infected cells contain HIV-1 provirus in a latent or defective form that was not detected in some earlier studies.