OBJECTIVES A recently released new QuantiFERON (QFT) product, QFT TB Gold plus (QFT-plus), is optimized for both CD4 and CD8 responses and reported to have higher sensitivity compared to the former QFT-3 G. Previously, using supernatants of QFT-3 G, we and others have demonstrated that cytokines other than IFN-γ may be useful in diagnosing tuberculosis. The present study aimed to identify cytokines that are useful for accurately diagnosing active tuberculosis by using QFT-plus and compared the data to those with QFT-3 G. METHODS Eighty-three active tuberculosis patients and 70 healthy control subjects who were examined by QFT at Tokyo National Hospital from June 2017 to July 2018 were enrolled. QFT-3 G and QFT-plus were performed according to the manufacturer's instructions. At the same time, blood cell culture supernatants were collected and assayed for their cytokine levels using R&D Systems Luminex Assay and MAGPIX System. The levels of cytokines were compared between different antigen-containing tubes (3 G Ag, TB1 and TB2 tubes), as well as between the patients and the control subjects. ROC curves were drawn, and the AUCs were calculated. RESULTS Five cytokines, i.e., IL-2, IL-6, IL-8, IP-10 and MIP-1β, produced by human blood cells in three independent tubes containing different tuberculosis antigens were higher in the 3 G Ag tube compared to both the TB1 and TB2 tubes. Further, when the TB1 and TB2 tubes were compared, TB2 showed greater production of only PDGF-BB, and less production of IL-6 and TNF-α. For diagnosing active tuberculosis, the levels of IP-10 were superior to the level of IFN-γ based on showing a larger AUC for ROC curves in our present study setting. Finally, the levels of IFN-γ, IL-1RA, IL-2, IP-10, MCP-1 and MIP-1β were distinctly different between the active tuberculosis patients and healthy controls. CONCLUSIONS In summary, there was no cytokine that was higher in the tubes of QFT-plus compared to the tube of QFT-3 G, suggesting inferiority of QFT-plus antigens to 3 G Ag in terms of elicitation of cytokine production. Our results also suggest the usefulness of cytokines that showed a significant difference between the active tuberculosis patients and the healthy controls-namely, IFN-γ, IL-1RA, IL-2, IP-10, MCP-1 and MIP-1β-for diagnosing tuberculosis, but the roles of these cytokines in the pathogenesis of tuberculosis need to be elucidated (UMIN000035253).

中文翻译：

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使用QuantiFERON-TB Gold Plus对活动性肺结核患者进行细胞因子水平评估。

目的最近发布的一种新的QuantiFERON（QFT）产品QFT TB Gold plus（QFT-plus）已针对CD4和CD8响应进行了优化，并据报道比以前的QFT-3 G具有更高的灵敏度。以前，使用QFT的上清液-3 G，我们和其他人已证明，IFN-γ以外的其他细胞因子可能在诊断结核中有用。本研究旨在通过QFT-plus识别可用于准确诊断活动性肺结核的细胞因子，并将数据与QFT-3 G进行比较。方法83例活动性结核病患者和70例健康对照受试者在接受QFT时接受了检查东京国立医院于2017年6月至2018年7月入组。QFT-3 G和QFT-plus根据制造商的说明进行。同时，收集血细胞培养上清液，并使用R＆D Systems Luminex Assay和MAGPIX System测定其细胞因子水平。比较了不同的含抗原试管（3 G Ag，TB1和TB2管）之间以及患者与对照组之间的细胞因子水平。绘制ROC曲线，并计算AUC。结果人血细胞在含有不同结核抗原的三个独立试管中由人血细胞产生的五种细胞因子，即IL-2，IL-6，IL-8，IP-10和MIP-1β在3 G Ag管中比两者均高TB1和TB2管。此外，当比较TB1和TB2管时，TB2仅显示PDGF-BB的产量更高，而IL-6和TNF-α的产量更低。为了诊断活动性肺结核，IP-10水平优于IFN-γ水平，因为在我们目前的研究环境中，ROC曲线显示出较大的AUC。最后，在活动性结核病患者和健康对照组之间，IFN-γ，IL-1RA，IL-2，IP-10，MCP-1和MIP-1β的水平明显不同。结论综上所述，在QFT-plus试管中，没有比QFT-3 G试管中更高的细胞因子，这表明在诱导细胞因子产生方面，QFT-plus抗原比3 G Ag低。我们的研究结果还表明，在活动性结核病患者和健康对照之间显示出显着差异的细胞因子的有用性，即IFN-γ，IL-1RA，IL-2，IP-10，MCP-1和MIP-1β-诊断结核病，但需要阐明这些细胞因子在结核病发病机理中的作用（UMIN000035253）。