Microculture assay for isolation of human immunodeficiency virus type 1 and for titration of infected peripheral blood mononuclear cells.
Microculture assay for isolation of human immunodeficiency virus type 1 and for titration of infected peripheral blood mononuclear cells.
复制标题
用于分离 1 型人类免疫缺陷病毒和滴定感染的外周血单核细胞的微量培养测定。
DOI：
10.1128/jcm.28.4.734-737.1990
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发表时间：
1990
影响因子：
9.4
通讯作者：
Hollinger,FB
中科院分区：
文献类型：
--
作者：
Dimitrov,DH;Melnick,JL;Hollinger,FB
为了确定人类免疫缺陷病毒（HIV）在微培养中检测的最佳条件，用不同比例的患者和供者外周血单个核细胞（PBMCs）进行了实验。供体/患者PBMC比例为1:1至1:125。当150万个供体细胞与等量或更少量的患者pbmc共培养时，获得最佳结果。因此，可以通过稀释患者细胞来达到病毒学终点。供体细胞数量较少，患者细胞数量较多或不较多，导致HIV分离率较低。同样，在不添加正常供体细胞的情况下，用植物血凝素直接刺激患者外周血单核细胞也会显著降低检测的敏感性。我们建议，使用一定量的供体细胞和不同稀释度的患者细胞进行微培养，可能有助于监测抗病毒治疗期间HIV水平的变化。
To define the optimal conditions for human immunodeficiency virus (HIV) detection in microcultures, experiments were conducted with different ratios of patient and donor peripheral blood mononuclear cells (PBMCs). Donor/patient PBMC ratios ranged from 1:1 to 1:125. Optimal results were obtained when 1,500,000 donor cells were cocultured with equal or smaller quantities of patient PBMCs. Thus, virologic endpoints could be achieved by diluting patient cells. Smaller numbers of donor cells, with or without larger numbers of patients cells, resulted in lower rates of HIV isolation. Similarly, the direct stimulation of patient PBMCs with phytohemagglutinin without the addition of normal donor cells lowered the sensitivity of the assay significantly. We suggest that a microculture procedure using a fixed quantity of donor cells with different dilutions of patient cells may be useful for monitoring changing HIV levels during antiviral therapy.