A resting cell assay for improved detection of antibody-mediated neutralization of HIV type 1 primary isolates.
A resting cell assay for improved detection of antibody-mediated neutralization of HIV type 1 primary isolates.
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一种静息细胞测定，用于改进抗体介导的 HIV 1 型初级分离株中和的检测。
DOI：
10.1089/aid.1995.11.1449
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Sharpe,S
中科院分区：
文献类型：
--
作者：
Zolla-Pazner,S;Sharpe,S
在体外检测抗体介导的中和作用的灵敏度取决于病毒、抗体、靶细胞和检测中使用的培养条件。利用活化和转化的靶细胞，深入研究了不同培养适应株和原代HIV-1毒株被不同的多克隆和单抗制剂中和的能力。然而，绝大多数活体中对HIV-1敏感的CD4+细胞并没有被激活或转化，而是处于静止状态。由于静息淋巴细胞可以感染HIV-1，我们启动了一些研究，以确定(1)使用静息淋巴细胞作为靶细胞是否会导致中和试验的灵敏度提高，(2)靶细胞的激活程度是否对检测抗体介导的中和作用的灵敏度有可测量的影响，以及(3)使用更灵敏的分析方法，患者血清中的中和抗体是否可能被检测到，而使用“常规”分析时，中和抗体的检测低于检测阈值。以下研究中描述的实验表明，靶细胞激活水平与检测中和的灵敏度之间存在相反的关系。此外，使用一种以未刺激的外周血单核细胞为靶细胞的检测方法，实验表明，与以刺激的外周血单核细胞为靶细胞时相比，抗体介导的HIV-1原代和实验室原型分离株的敏感性可以提高10到100倍。利用这种静息细胞检测方法，可以在HIV阳性受试者的血清中检测到中和活性，而以前使用的“常规”中和检测方法是无法检测到的。
The sensitivity with which antibody-mediated neutralization is detectedin vitrois dependent on the virus, the antibody, the target cells, and the culture conditions used in the assay. Using activated and transformed target cells, the ability of various culture-adapted and primary strains of HIV-1 to be neutralized by different polyclonal and monoclonal antibody preparations has been thoroughly studied. However, the vast majority of HIV-1-susceptible CD4+cellsin vivoare not activated or transformed, but are quiescent. Because resting lymphocytes can be infected with HIV-1, we initiated studies to determine (1) if the use of resting lymphocytes as target cells would result in a neutralization assay with increased sensitivity, (2) if the degree of target cell activation had a measurable effect on the sensitivity with which antibody-mediated neutralization could be detected, and (3) whether, using a more sensitive assay, neutralizing antibodies in patients' sera might be detectable that had been below the threshold of detection when using "conventional" assays. The experiments described in the studies below reveal that an inverse relationship exists between the level of target cell activation and the sensitivity with which neutralization can be detected. Moreover, using an assay in which unstimulated peripheral blood mononuclear cells serve as target cells, experiments show that antibody-mediated neutralization of primary and prototype laboratory isolates of HIV-1 can be detected with 10- to 100-fold greater sensitivity than when stimulated peripheral blood mononuclear cells are used as target cells. With this resting cell assay, neutralizing activity can be detected in the sera of HIV-positive subjects that, by previously used "conventional" neutralization assays, was undetectable.
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影响因子：
4.4
作者：
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影响因子：
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影响因子：
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作者：
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通讯作者：
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影响因子：
4.4
作者：
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影响因子：
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作者：
WRIN, T;LOH, TP;NUNBERG, JH
通讯作者：
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