Seshadri Anupamaa, Brat Gabriel A, Yorkgitis Brian K, Keegan Joshua, Dolan James, Salim Ali, Askari Reza, Lederer James A
All authors: Department of Surgery, Brigham and Women's Hospital and Harvard Medical School, Boston, MA.
Crit Care Med. 2017 Sep;45(9):1523-1530. doi: 10.1097/CCM.0000000000002577.
Trauma induces a complex immune response that requires a systems biology research approach. Here, we used a novel technology, mass cytometry by time-of-flight, to comprehensively characterize the multicellular response to trauma.
Peripheral blood mononuclear cells samples were stained with a 38-marker immunophenotyping cytometry by time-of-flight panel. Separately, matched peripheral blood mononuclear cells were stimulated in vitro with heat-killed Streptococcus pneumoniae or CD3/CD28 antibodies and stained with a 38-marker cytokine panel. Monocytes were studied for phagocytosis and oxidative burst.
Single-institution level 1 trauma center.
Trauma patients with injury severity scores greater than 20 (n = 10) at days 1, 3, and 5 after injury, and age- and gender-matched controls.
None.
Trauma-induced expansion of Th17-type CD4 T cells was seen with increased expression of interleukin-17 and interleukin-22 by day 5 after injury. Natural killer cells showed reduced T-bet expression at day 1 with an associated decrease in tumor necrosis factor-β, interferon-γ, and monocyte chemoattractant protein-1. Monocytes showed robust expansion following trauma but displayed decreased stimulated proinflammatory cytokine production and significantly reduced human leukocyte antigen - antigen D related expression. Further analysis of trauma-induced monocytes indicated that phagocytosis was no different from controls. However, monocyte oxidative burst after stimulation increased significantly after injury.
Using cytometry by time-of-flight, we were able to identify several major time-dependent phenotypic changes in blood immune cell subsets that occur following trauma, including induction of Th17-type CD4 T cells, reduced T-bet expression by natural killer cells, and expansion of blood monocytes with less proinflammatory cytokine response to bacterial stimulation and less human leukocyte antigen - antigen D related. We hypothesized that monocyte function might be suppressed after injury. However, monocyte phagocytosis was normal and oxidative burst was augmented, suggesting that their innate antimicrobial functions were preserved. Future studies will better characterize the cell subsets identified as being significantly altered by trauma using cytometry by time-of-flight, RNAseq technology, and functional studies.
创伤会引发复杂的免疫反应，这需要采用系统生物学研究方法。在此，我们使用了一种新技术——飞行时间质谱流式细胞术，来全面表征对创伤的多细胞反应。
外周血单个核细胞样本用一个包含38种标志物的飞行时间免疫表型流式细胞术检测板进行染色。另外，将匹配的外周血单个核细胞在体外分别用热灭活的肺炎链球菌或CD3/CD28抗体刺激，并用一个包含38种标志物的细胞因子检测板进行染色。对单核细胞进行吞噬作用和氧化爆发的研究。
单机构一级创伤中心。
受伤严重程度评分大于20分的创伤患者（n = 10），分别在受伤后第1天、第3天和第5天，以及年龄和性别匹配的对照组。
无。
创伤后诱导了Th17型CD4 T细胞的扩增，在受伤后第5天时白细胞介素-17和白细胞介素-22的表达增加。自然杀伤细胞在第1天时T-bet表达降低，同时肿瘤坏死因子-β、干扰素-γ和单核细胞趋化蛋白-1也相应减少。单核细胞在创伤后出现强劲扩增，但刺激后的促炎细胞因子产生减少，且人白细胞抗原-D相关表达显著降低。对创伤诱导的单核细胞的进一步分析表明，其吞噬作用与对照组无差异。然而，受伤后刺激后的单核细胞氧化爆发显著增加。
通过飞行时间质谱流式细胞术，我们能够识别创伤后血液免疫细胞亚群中几种主要的时间依赖性表型变化，包括Th17型CD4 T细胞的诱导、自然杀伤细胞T-bet表达降低，以及血液单核细胞扩增，其对细菌刺激的促炎细胞因子反应减弱且人白细胞抗原-D相关表达减少。我们推测受伤后单核细胞功能可能受到抑制。然而，单核细胞吞噬作用正常且氧化爆发增强，表明其天然抗菌功能得以保留。未来的研究将利用飞行时间质谱流式细胞术、RNA测序技术和功能研究，更好地表征被确定为因创伤而发生显著改变的细胞亚群。
