Orlov Marika, Dmyterko Victoria, Wurfel Mark M, Mikacenic Carmen
Department of Medicine, University of Washington, Seattle, Washington, United States of America.
Department of Medicine, Division of Pulmonary and Critical Care Medicine, University of Washington, Seattle, Washington, United States of America.
PLoS One. 2017 Aug 14;12(8):e0182966. doi: 10.1371/journal.pone.0182966. eCollection 2017.
CD4+ T-helper 17 (Th17) cells and Interleukin (IL)-17A play an important role in clearing pathogens in mouse models of pneumonia. We hypothesized that numbers of Th17 cells and levels of IL-17A are associated with risk for nosocomial pneumonia in humans.
We collected bronchoalveolar lavage (BAL) fluid from mechanically ventilated (n = 25) patients undergoing quantitative bacterial culture to evaluate for ventilator associated pneumonia (VAP). We identified Th17 cells by positive selection of CD4+ cells, stimulation with ionomycin and PMA, then staining for CD4, CD45, CCR6, IL-17A, and IFN-γ followed by flow cytometric analysis (n = 21). We measured inflammatory cytokine levels, including IL-17A, in BAL fluid by immunoassay.
VAP was detected in 13 of the 25 subjects. We identified a decreased percentage of IL-17A producing Th17 cells in BAL fluid from patients with VAP compared to those without (p = 0.02). However, we found no significant difference in levels of IL-17A in patients with VAP compared to those without (p = 0.07). Interestingly, IL-17A levels did not correlate with Th17 cell numbers. IL-17A levels did show strong positive correlations with alveolar neutrophil numbers and total protein levels.
Th17 cells are found at lower percentages in BAL fluid from mechanically ventilated patients with VAP and IL-17A levels correlated with Th17 cell percentages in non-VAP subjects, but not those with VAP. These findings suggest that Th17 cells may be protective against development of nosocomial pneumonia in patients receiving mechanical ventilation and that alveolar IL-17A in VAP may be derived from sources other than alveolar Th17 cells.
在肺炎小鼠模型中,CD4 +辅助性T细胞17(Th17)和白细胞介素(IL)-17A在清除病原体方面发挥重要作用。我们推测Th17细胞数量和IL-17A水平与人类医院获得性肺炎的风险相关。
我们收集了接受定量细菌培养以评估呼吸机相关性肺炎(VAP)的机械通气患者(n = 25)的支气管肺泡灌洗(BAL)液。我们通过阳性选择CD4 +细胞、用离子霉素和佛波酯刺激,然后对CD4、CD45、CCR6、IL-17A和IFN-γ进行染色,随后进行流式细胞术分析来鉴定Th17细胞(n = 21)。我们通过免疫测定法测量BAL液中的炎性细胞因子水平,包括IL-17A。
25名受试者中有13名检测到VAP。我们发现与未患VAP的患者相比,VAP患者BAL液中产生IL-17A的Th17细胞百分比降低(p = 0.02)。然而,我们发现VAP患者与未患VAP患者的IL-17A水平没有显著差异(p = 0.07)。有趣的是,IL-17A水平与Th17细胞数量无关。IL-17A水平与肺泡中性粒细胞数量和总蛋白水平呈强正相关。
在发生VAP的机械通气患者的BAL液中,Th17细胞的百分比更低,并且IL-17A水平与非VAP受试者的Th17细胞百分比相关,但与VAP患者无关。这些发现表明Th17细胞可能对接受机械通气的患者发生医院获得性肺炎具有保护作用,并且VAP中的肺泡IL-17A可能来源于肺泡Th17细胞以外的其他来源。