Modification of the HCMV-specific IFN-γ release test (QuantiFERON-CMV) and a novel proposal for its application
Modification of the HCMV-specific IFN-γ release test (QuantiFERON-CMV) and a novel proposal for its application
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DOI：
10.5387/fms.2017-01
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发表时间：
2017-08-01
影响因子：
0.8
通讯作者：
Suzutani, Tatsuo
中科院分区：
文献类型：
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作者：
Kobayashi, Takahiro;Sato, Jun-Ichi;Suzutani, Tatsuo
人巨细胞病毒（Human cytomegalovirus，HCMV）是一种广泛分布于人体内的病毒，对人体无不良影响，但对器官移植受者、艾滋病患者等免疫功能低下的患者却有严重的致病作用。为了管理这些免疫功能低下的患者，需要进行简单的临床检查以监测疾病风险。在这项研究中，我们使用HCMV立即早期-1（IE-1）或pp 65全蛋白或UV灭活的HCMV颗粒作为抗原，修改了干扰素-γ（IFN-γ）释放试验（QuantiFERON（R）CMV）。来自健康志愿者的肝素化外周血对pp 65蛋白的反应显示出明显的剂量依赖性S形曲线，尽管在该测定和ELISPOT测定的结果之间没有观察到相关性。发现向血液样品中添加pp 65的最终浓度为1x 10（3）至1x 10（5）pg/ml是最佳的。使用这种检测方法，我们观察到志愿者每天摄入酸奶8周后细胞免疫力显著增强，这表明除了监测HCMV感染风险外，该检测方法还具有新的应用。在HCMV抗原刺激下，外周血细胞分泌的IFN-γ在个体之间存在显著差异;因此，该测定无法标准化。尽管如此，人们发现它对于观察个体水平上细胞免疫活性的波动特别有用。
Human cytomegalovirus (HCMV) is universally distributed among humans without any adverse effects; however it induces severe diseases in immunocompromised patients such as organ transplant recipients and AIDS patients. To manage these immunocompromised patients, an easy clinical examination for the monitoring of disease risk is required. In this study, we modified the interferon-gamma (IFN-gamma) release test (QuantiFERON (R) CMV) using HCMV immediate early-1 (IE-1) or pp65 whole proteins, or UV inactivated HCMV particles as an antigen. The response of heparinized peripheral blood from healthy volunteers to the pp65 protein showed an obvious dose-dependent sigmoid curve, although no correlation was observed between results of this assay and an ELISPOT assay. The addition of pp65 to the blood samples at a final concentration of 1x10(3) to 1x10(5) pg/ml was found to be optimum. Using this assay, we observed a significant enhancement in cellular immunity in volunteers after the daily ingestion of yogurt for 8 weeks, which suggested a novel application of the assay in addition to monitoring HCMV infection risk. IFN-gamma secretion from peripheral blood cells on HCMV-antigen stimulation differed significantly between individuals ; therefore, the assay could not be normalized. Nevertheless, it was found to be particularly useful for observing fluctuations in cellular immune activity on an individual level.