Department of Immunology and Infectious Biology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland.
Department of Immunology and Infectious Biology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland; Department of Neurology and Stroke, Medical University of Lodz, Lodz, Poland.
Vaccine. 2018 Jul 16;36(30):4566-4577. doi: 10.1016/j.vaccine.2018.05.068. Epub 2018 Jun 19.
Bacillus Calmette-Guérin (BCG) is the only vaccine available against tuberculosis and the tuberculin skin test (TST) is the most widely used method to detect BCG take. However, subjects may remain TST-negative, even after several BCG administrations. To investigate some of the potential reasons underlying this inability of developing tuberculin sensitivity in response to BCG we compared the effect of different mycobacterial stimuli in the groups differently responding to tuberculin. TST was performed on 71 healthy adults aged 25-30 years, who had received BCG in their childhood, and considered TST-positive at ≥10 mm. Dendritic cells (DCs) were incubated with PPD, live BCG or rBCGhIL-18, producing human IL-18. The latter strain was used to investigate whether the production of IL-18 could overcome some of the immune read-out limitations in the TST-negative subjects. CD86, CD80, CD40, and DC-specific intracellular adhesion molecule-3 grabbing nonintegrin (DC-SIGN) expression was analysed by flow cytometry and IL-10, IL-23 and IP-10 secretion in culture supernatants by ELISA. In DCs-T cell co-cultures with naive and memory CD4 T cells, the IFN-γ and IL-10 levels were determined by ELISA. We found no difference in IL-10 and IFN-γ production by naive T cells between the TST-negative and TST-positive subjects. However, IFN-γ was produced in significantly higher amounts by memory T cells incubated with PPD, BCG or rBCGhIL-18-pulsed DCs in TST-positive than in TST-negative subjects, whereas the numbers of the IFN-γ-producing T cells were similar in both groups. This difference may be partially due to a decreased CD40 and enhanced reduction in DC-SIGN expression by DCs of TST-negative versus TST-positive subjects. A strong effect of IL-18 expression by rBCGhIL-18 on IL-23 production by the DC was seen in both groups, which likely was the reason for the increased IFN-γ production by naïve T cells upon incubation with mycobacteria-pulsed DC, regardless of the TST status.
卡介苗（BCG）是唯一可用的结核病疫苗，结核菌素皮肤试验（TST）是最广泛用于检测卡介苗接种的方法。然而，即使多次接种 BCG，仍有一些受试者 TST 呈阴性。为了研究导致这种对卡介苗产生结核敏感性能力缺失的一些潜在原因，我们比较了不同分枝杆菌刺激物在对结核菌素反应不同的组中的作用。对 71 名年龄在 25-30 岁之间的健康成年人进行了 TST，这些成年人在儿童时期接种过卡介苗，并且 TST 结果≥10mm 为阳性。将树突状细胞（DC）与 PPD、活卡介苗或 rBCGhIL-18 孵育，产生人 IL-18。后一种菌株用于研究 IL-18 的产生是否可以克服 TST 阴性受试者中一些免疫检测的局限性。通过流式细胞术分析 CD86、CD80、CD40 和 DC 特异性细胞间黏附分子-3 抓取非整合素（DC-SIGN）的表达，通过 ELISA 检测培养上清液中 IL-10、IL-23 和 IP-10 的分泌。在幼稚和记忆 CD4 T 细胞的 DC-T 细胞共培养物中，通过 ELISA 测定 IFN-γ和 IL-10 水平。我们发现，TST 阴性和 TST 阳性受试者之间，幼稚 T 细胞产生的 IL-10 和 IFN-γ 没有差异。然而，与 TST 阴性受试者相比，用 PPD、BCG 或 rBCGhIL-18 脉冲 DC 孵育的记忆 T 细胞产生的 IFN-γ 明显更多，而两组中产生 IFN-γ 的 T 细胞数量相似。这种差异可能部分归因于 TST 阴性与 TST 阳性受试者的 DC 中 CD40 减少和 DC-SIGN 表达增强。rBCGhIL-18 对 DC 中 IL-23 产生的强烈影响在两组中均可见，这可能是无论 TST 状态如何，用分枝杆菌脉冲 DC 孵育后幼稚 T 细胞 IFN-γ 产生增加的原因。
