Neal T F, Holland H K, Baum C M, Villinger F, Ansari A A, Saral R, Wingard J R, Fleming W H
Department of Medicine, Emory University, Atlanta, GA 30322, USA.
Blood. 1995 Sep 1;86(5):1749-56.
Controversy exists as to whether hematopoietic progenitor cells are infected by human immunodeficiency virus-1 (HIV-1) in vivo. Most studies have focused on patients with acquired immunodeficiency syndrome (AIDS)/AIDS-related complex, and little data are available on asymptomatic patients with well preserved CD4+ T-cell counts. To determine if CD34+ hematopoietic progenitor cells are infected early in the course of HIV-1 disease, we evaluated 10 asymptomatic HIV-1 seropositive (HIV-1+) patients. The CD34+ cell fraction was purified by a two-step procedure consisting of both affinity chromatography and fluorescence-activated cell sorting that resulted in a median purity of over 99%. Using conventional and nested polymerase chain reaction (PCR) assays, we evaluated the presence and frequency of HIV-1 proviral DNA. Both bone marrow mononuclear cells and CD34- cells from all 10 patients were strongly positive for the HIV-1 pol and/or gag gene sequences. In contrast, sorted CD34+ cells from only two of 10 patients were positive, and the number of copies of proviral DNA in these samples was estimated to be from 2 to 5 per 250,000 cells. To test the in vitro functional capacity of CD34+ progenitors, these cells were assayed in both methylcellulose and long-term stromal culture. We found no significant reduction in the number of colony-forming unit-erythroid (CFU-E), burst-forming unit-erythroid (BFU-E), or colony-forming unit-granulocyte macrophage (CFU-GM) colonies, or in the frequency of cobblestone area forming cells from limit dilution analysis in HIV-1+ asymptomatic patients. Pooled methylcellulose colonies generated from CD34+ cells were HIV-1- in nine of 10 samples. All progeny from long-term cultures of CD34+ cells were HIV-1-. We conclude that the CD34+ hematopoietic progenitor compartment is not infected in the majority of asymptomatic HIV-1+ patients, and that these cells may represent a suitable target for strategies designed to protect developing CD4+ T cells from infection.
造血祖细胞在体内是否会被人类免疫缺陷病毒1型（HIV-1）感染存在争议。大多数研究聚焦于获得性免疫缺陷综合征（AIDS）/AIDS相关综合征患者，而关于CD4+T细胞计数保存良好的无症状患者的数据很少。为了确定CD34+造血祖细胞是否在HIV-1疾病进程早期被感染，我们评估了10名无症状HIV-1血清阳性（HIV-1+）患者。通过亲和层析和荧光激活细胞分选两步法纯化CD34+细胞组分，最终纯度中位数超过99%。使用常规和巢式聚合酶链反应（PCR）检测，我们评估了HIV-1前病毒DNA的存在情况和频率。所有10名患者的骨髓单个核细胞和CD34-细胞的HIV-1 pol和/或gag基因序列均呈强阳性。相比之下，10名患者中只有2名患者分选的CD34+细胞呈阳性，这些样本中前病毒DNA的拷贝数估计为每250,000个细胞中有2至5个。为了测试CD34+祖细胞的体外功能能力，在甲基纤维素和长期基质培养中对这些细胞进行了检测。我们发现，在HIV-1+无症状患者中，红系集落形成单位（CFU-E）、红系爆式集落形成单位（BFU-E）或粒巨噬系集落形成单位（CFU-GM）集落的数量，或通过极限稀释分析得出的鹅卵石区域形成细胞的频率均无显著降低。从CD34+细胞产生的合并甲基纤维素集落在10个样本中有9个为HIV-1阴性。CD34+细胞长期培养的所有子代均为HIV-1阴性。我们得出结论，在大多数无症状HIV-1+患者中，CD34+造血祖细胞区未被感染，并且这些细胞可能是旨在保护发育中的CD4+T细胞免受感染的策略的合适靶点。
