Development of an HSV-1 neutralization test with a glycoprotein D specific antibody for measurement of neutralizing antibody titer in human sera.
Development of an HSV-1 neutralization test with a glycoprotein D specific antibody for measurement of neutralizing antibody titer in human sera.
复制标题
使用糖蛋白 D 特异性抗体开发 HSV-1 中和测试，用于测量人血清中的中和抗体滴度
DOI：
10.1186/s12985-016-0508-4
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发表时间：
2016-03-18
期刊：
影响因子：
4.8
通讯作者：
Xia NS
中科院分区：
文献类型：
--
作者：
Luo Y;Xiong D;Li HH;Qiu SP;Lin CL;Chen Q;Huang CH;Yuan Q;Zhang J;Xia NS
研究背景检测HSV-1中和抗体（NAb）滴度对监测易感人群对HSV-1的免疫保护作用至关重要，这将有助于开发抗HSV-1感染的疫苗和改进以HSV-1为基础的溶瘤病毒治疗。我们开发了基于酶联免疫斑点试验的中和试验通过ELISA（ELISPOT-NT）测定人血清样品中抗HSV-1的中和抗体滴度。该优化的测定采用特异性识别糖蛋白D的单克隆抗体来检测HSV-1感染的细胞。通过该试验，可在一天内通过自动判读细胞斑点计数来测定抗HSV-1的中和抗体滴度。我们通过检测22份代表不同滴度的人血清样品，观察到ELISPOT-NT和空斑减少中和试验（PRNT）所得结果具有良好的相关性。此外，对从不同年龄组收集的269份人血清样本进行了检测，（log ~ 2NT_（50））为8.3 ± 2.8，NAb检出率为83.6%，不同年龄组平均中和抗体滴度随年龄增长而升高，结论该方法简便、快速、准确，可用于HSV-1中和抗体滴度的检测。1在大型队列中。
BackgroundInvestigating the neutralizing antibody (NAb) titer against HSV-1 is essential for monitoring the immune protection against HSV-1 in susceptible populations, which would facilitate the development of vaccines against herpes infection and improvement of HSV-1 based oncolytic virotherapy.ResultsIn this study, we have developed a neutralization test based on the enzyme-linked immunospot assay (ELISPOT-NT) to determine the neutralizing antibody titer against HSV-1 in human serum samples. This optimized assay employed a monoclonal antibody specifically recognizing glycoprotein D to detect the HSV-1 infected cells. With this test, the neutralizing antibody titer against HSV-1 could be determined within one day by automated interpretation of the counts of cell spots. We observed good correlation in the results obtained from ELISPOT-NT and plaque reduction neutralization test (PRNT) by testing 22 human serum samples representing different titers. Moreover, 269 human serum samples collected from a wide range of age groups were tested, the average neutralizing antibody titer (log2NT50) was 8.3 ± 2.8 and the prevalence of NAbs was 83.6 % in this cohort, it also revealed that the average neutralizing antibody titer in different groups increased with the age, and no significant difference in neutralizing antibody titers was observed between males and females.ConclusionsThese results prove that this novel assay would serve as an accurate and simple assay for the assessment of the neutralizing antibody titers against HSV-1 in large cohorts.