Neutralization sensitivity of cell culture passaged simian immunodeficiency virus
Neutralization sensitivity of cell culture passaged simian immunodeficiency virus
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DOI：
10.1128/jvi.71.10.7895-7902.1997
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发表时间：
1997-10-01
影响因子：
5.4
通讯作者：
Desrosiers, RC
中科院分区：
文献类型：
--
作者：
Means, RE;Greenough, T;Desrosiers, RC
构建了基于CEMx174和c8166 -45的细胞系，这些细胞系含有分泌的碱性磷酸酶（SEAP）报告基因，并受到来自SIVmac239或HIV-1（NL4-3）的应答启动子的控制。这些细胞系的SEAP活性基础水平较低，但转染该表达质粒后，SEAP活性大大提高。这些细胞系感染猴免疫缺陷病毒(SIS？人类免疫缺陷病毒1型（HIV-1）导致SEAP产量在48至72小时内急剧增加，这与感染病毒的数量直接相关。当将这些细胞与无细胞上清中SEAP活性的化学发光测量相结合时，这些细胞形成了快速、敏感和定量检测SIV和HIV感染和中和的基础。8个HIV-I原代分离株中有8个在该系统中诱导了易于测量的SEAP活性。虽然用其他方法很难检测到克隆的STVmac239的血清中和作用，但这种新的检测系统很容易以低滴度检测到SIVmac239的中和作用。用感染siv的猴子血清检测了两种不同细胞培养传代史的SIVmac251的中和敏感性。SIVmac251原代代仅在恒河猴外周血单核细胞原代培养中传代两次，而实验室适应的原代代已在MT4永生化t细胞系中广泛传代。SIVmac251的原株比实验室适应病毒更能抵抗siv感染猴子的多克隆血清的中和作用。因此，SIVmac似乎与HIV-1相似，通过t细胞系进行广泛的实验室传递，导致病毒对血清中和更加敏感。
CEMx174- and C8166-45-based cell lines which contain a secreted alkaline phosphatase (SEAP) reporter gene under the control of a tat-responsive promoter derived from either SIVmac239 or HIV-1(NL4-3) were constructed. Basal levels of SEAP activity from these cell lines were low but were greatly stimulated upon transfection of tat expression plasmids. Infection of these cell lines with simian immunodeficiency virus (SIS? or human immunodeficiency virus type 1 (HIV-1) resulted in a dramatic increase in SEAP production within 48 to 72 h that directly correlated with the amount of infecting virus. When combined,vith chemiluminescent measurement of SEAP activity in the cell-free supernatant, these cells formed the basis of a rapid, sensitive, and quantitative assay for SIV and HIV infectivity and neutralization. Eight of eight primary isolates of HIV-I that were tested induced readily measurable SEAP activity in this system. While serum neutralization of cloned STVmac239 was difficult to detect with other assays, neutralization of SIVmac239 was readily detected at low titers with this new assay system. The neutralization sensitivities of two stocks of SIVmac251 with different cell culture passage histories were tested by using sera from SIV-infected monkeys. The primary stock of SIVmac251 had been passaged only twice through primary cultures of rhesus monkey peripheral blood mononuclear cells, while the laboratory-adapted stock had been extensively passaged through the MT4 immortalized T-cell line. The primary stock of SIVmac251 was much more resistant to neutralization by a battery of poly clonal sera from SIV-infected monkeys than was the laboratory-adapted virus. Thus, SIVmac appears to be similar to HIV-1 in that extensive laboratory passage through T-cell lines resulted in a virus that is much more sensitive to serum neutralization.