Xin X, Shioda T, Fukushima M, Hu H, Oka S, Iwamoto A, Nagai Y
Department of Viral Infection, University of Tokyo, Japan.
Arch Virol. 1998;143(1):85-95. doi: 10.1007/s007050050270.
We previously reported that infectivities of human and other non-human lentiviruses including human immunodeficiency virus type 1 (HIV-1) are activated by desialylation of the virion surface [H. Hu et al., J Virol 70: 7,462-7,470 (1996)]. The present study was designed to determine whether neuraminidase (NA) is useful for isolation of HIV-1 from patients. Peripheral blood mononuclear cells (PBMC) or CD4+ cells isolated from the PBMC of infected individuals were cocultured with PBMC or CD4+ cells from uninfected healthy donors, and the efficiency and frequency of virus isolation in the presence of NA were compared with those by a routine conventional procedure. In a total of 41 isolation trials from 28 individuals, the presence of NA markedly increased the frequency of isolation. Furthermore, both the day when virus was first detected and the day when the virus titer was the highest in the cultures were significantly earlier in the presence of NA than in the absence of NA. The deduced amino acid sequences of the V2 and V3 regions of gp120 were identical or very similar between the isolates obtained in the presence or absence of NA, suggesting that both isolation procedures selected a similar population. NA was thus found to facilitate HIV-1 isolation and its use is recommended particularly when isolation is negative by the conventional procedures.
我们之前报道过,包括人类免疫缺陷病毒1型(HIV-1)在内的人类及其他非人类慢病毒的感染性可通过病毒粒子表面的去唾液酸化作用被激活[H. Hu等人,《病毒学杂志》70: 7462 - 7470(1996年)]。本研究旨在确定神经氨酸酶(NA)是否有助于从患者中分离HIV-1。将从受感染个体的外周血单核细胞(PBMC)中分离出的PBMC或CD4⁺细胞与未感染健康供体的PBMC或CD4⁺细胞进行共培养,并将存在NA时病毒分离的效率和频率与常规传统方法进行比较。在对28名个体进行的总共41次分离试验中,NA的存在显著提高了分离频率。此外,在存在NA的情况下,培养物中首次检测到病毒的日期以及病毒滴度最高的日期均明显早于不存在NA的情况。在存在或不存在NA的情况下获得的分离株之间,gp120的V2和V3区域推导的氨基酸序列相同或非常相似,这表明两种分离程序选择了相似的群体。因此发现NA有助于HIV-1的分离,特别在常规程序分离为阴性时,建议使用NA。