Department of Paediatrics, "Sapienza" University of Rome, Rome Italy.
Department of Infectious Diseases, Istituto Superiore di Sanità, Rome, Italy.
J Biol Regul Homeost Agents. 2018 Sep-Oct;32(5):1339-1344.
The aim of the present study was the development of a reliable method to evaluate the pattern of the ongoing T-cell response in young infants affected by respiratory infection. To this purpose, we enrolled 44 infants hospitalized with a diagnosis of respiratory syncytial virus bronchiolitis. After a short-term stimulation of whole blood samples, intracellular IFN-g and IL-4 cytokines were measured in CD4+ and CD8+ T-cell subsets by flow cytometry. A stringent staining and gating strategy was used in order to maximize the reduction of background noise and to exclude false positives. The frequencies of cytokine-producing T-cell subsets, albeit low, were easily quantifiable. Cytokine responses were higher in infants sampled > 7 days from the onset of symptoms. The use of a rigorous strategy for cell staining and gating, coupled with a short-term stimulation of whole blood and a careful evaluation of time elapsed from the onset of symptoms constitutes a convincing approach for future clinical studies.
本研究旨在开发一种可靠的方法，以评估受呼吸道感染影响的婴幼儿中持续 T 细胞反应的模式。为此，我们招募了 44 名因呼吸道合胞病毒细支气管炎住院的婴儿。在对全血样本进行短期刺激后，通过流式细胞术测量 CD4+和 CD8+T 细胞亚群中的 IFN-γ和 IL-4 细胞因子。使用严格的染色和门控策略，以最大限度地减少背景噪音并排除假阳性。细胞因子产生 T 细胞亚群的频率虽然较低，但很容易定量。在症状发作后 >7 天取样的婴儿中，细胞因子反应更高。使用严格的细胞染色和门控策略，结合全血的短期刺激以及从症状发作到评估时间的仔细评估，是未来临床研究的一种有说服力的方法。
