Kobayashi Takahiro, Sato Jun-Ichi, Ikuta Kazufumi, Kanno Ryoko, Nishiyama Kyoko, Koshizuka Tetsuo, Ishioka Ken, Suzutani Tatsuo
Department of Microbiology, Fukushima Medical University School of Medicine, Fukushima.
Division of Microbiology, Tohoku Medical and Pharmaceutical University.
Fukushima J Med Sci. 2017 Aug 9;63(2):64-74. doi: 10.5387/fms.2017-01. Epub 2017 Jun 22.
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-γ (IFN-γ) release test (QuantiFERON-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 1×10 to 1×10 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-γ 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.
人类巨细胞病毒（HCMV）在人群中普遍存在且无任何不良影响；然而，它会在免疫功能低下的患者中引发严重疾病，如器官移植受者和艾滋病患者。为了治疗这些免疫功能低下的患者，需要一种简单的临床检查来监测疾病风险。在本研究中，我们使用HCMV立即早期蛋白-1（IE-1）或pp65全蛋白，或紫外线灭活的HCMV颗粒作为抗原，对干扰素-γ（IFN-γ）释放试验（QuantiFERON-CMV）进行了改进。健康志愿者的肝素化外周血对pp65蛋白的反应呈现出明显的剂量依赖性S形曲线，尽管该检测结果与ELISPOT检测结果之间未观察到相关性。发现以1×10至1×10 pg/ml的终浓度向血样中添加pp65是最佳的。使用该检测方法，我们观察到志愿者在每天摄入酸奶8周后细胞免疫有显著增强，这表明该检测方法除了监测HCMV感染风险外还有新的应用。HCMV抗原刺激后外周血细胞分泌的IFN-γ在个体之间差异显著；因此，该检测无法标准化。然而，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。
