A Comparison of Human Neutrophils Acquired from Four Experimental Models of Inflammation.
A Comparison of Human Neutrophils Acquired from Four Experimental Models of Inflammation.
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DOI：
10.1371/journal.pone.0165502
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
O'Brien AJ
中科院分区：
文献类型：
--
作者：
Maini AA;George MJ;Motwani MP;Day RM;Gilroy DW;O'Brien AJ
中性粒细胞功能缺陷与多种临床症状有关。几个模型被用来研究激活的人类中性粒细胞，类似于在炎症部位发现的那些。这些技术包括全血(WB)体外脂多糖(LPS)刺激和体内技术：斑疹病毒水泡、皮肤窗口和紫外线灭活大肠杆菌(UVKEc)皮内注射。从这些实验中获得的中性粒细胞从未被比较过。我们比较了每种技术中性粒细胞的激活状态，以确定用于人体研究的最佳模型。健康的男性志愿者随机接受四种技术中的一种(n=5/组)。内毒素：WB用1 ng/mlLPS刺激4h。斑泻素：0.1%斑泻素溶液12.5μL在24小时后引出单发水泡。皮肤窗口：4个6 mm的机械吸水泡，去除屋顶，并在窗户上放置渗出物收集室4小时后进行抽吸。UVKEc：1.5×107 UVKEc皮内注射。单个10 mm机械吸液水泡形成，4小时吸气。以非刺激性白蛋白为对照。流式细胞仪检测CD16、CD11b、CD54、CD62L和CD88的活化状态。用吞噬试验评价细胞功能状态。中性粒细胞激活模式在所有模型中都是相似的。中性粒细胞CD11b在所有模型中均升高，以UVKEC最明显(p<；0.0001)，CD54也升高，但仅在内毒素模型中显著(p=0.001)。CD62L在所有4个模型中均显著降低(p<；0.0001)，CD88在所有模型中也均被抑制。CD16在各模型中均无明显变化，中性粒细胞吞噬能力也无明显差异。总而言之，从每种技术获得的中性粒细胞在激活标记表达或吞噬能力方面没有显著差异。因此，我们认为全血刺激是对住院患者群体进行实验挑战的最佳模式。
Defects in neutrophil function have been implicated in a wide spectrum of clinical conditions. Several models are employed to study activated human neutrophils akin to those found at a site of inflammation. These include whole blood (WB) ex vivo stimulation with lipopolysaccharide (LPS) and in vivo techniques: cantharidin blister, skin windows and intra-dermal injection of UV-killed E.coli (UVKEc). Neutrophils obtained from these have never been compared. We compared the activation status of neutrophils from each technique in order to inform the optimal model for use in human studies. Healthy male volunteers were randomised to undergo one of the four techniques (n = 5/group). LPS: WB stimulated with 1ng/ml of LPS for 4 hours. Cantharidin: 12.5μl of 0.1% cantharidin elicited a single blister, aspirated at 24 hours. Skin windows: four 6mm mechanical-suction blisters created, de-roofed and an exudate-collection chamber placed over the windows for 4 hours before aspiration. UVKEc: 1.5 x 107 UVKEc injected intra-dermally. A single 10mm mechanical-suction blister formed and aspirated at 4 hours. Unstimulated WB used as the control. Flow cytometry was used to determine activation status using CD16, CD11b, CD54, CD62L and CD88. Functional status was assessed with a phagocytosis assay. The pattern of neutrophil activation was similar in all models. Neutrophil CD11b was elevated in all models, most markedly in UVKEc (p<0.0001), and CD54 was also elevated but only significant in the LPS model (p = 0.001). CD62L was significantly reduced in all 4 models (p<0.0001) and CD88 was also suppressed in all. There were no changes in CD16 in any model, neither was there any significant difference in the phagocytic capacity of the neutrophils. In summary, there are no significant differences in activation marker expression or phagocytic capacity in the neutrophils obtained from each technique. Therefore we believe whole blood stimulation is the best model in experimentally challenging inpatient populations.
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DOI：
10.1002/cjp2.43
发表时间：
2016-07
期刊：
The journal of pathology. Clinical research
影响因子：
--
作者：
Motwani MP;Flint JD;De Maeyer RP;Fullerton JN;Smith AM;Marks DJ;Gilroy DW
通讯作者：
Gilroy DW
影响因子：
32.4
作者：
Venturi, GM;Tu, LL;Tedder, TF
通讯作者：
Tedder, TF
影响因子：
7.6
作者：
Harbord MW;Marks DJ;Forbes A;Bloom SL;Day RM;Segal AW
通讯作者：
Segal AW
影响因子：
3.6
作者：
Binder, R;Kress, A;Kirschfink, M
通讯作者：
Kirschfink, M
影响因子：
5.1
作者：
BRIHEIM, G;COBLE, B;DAHLGREN, C
通讯作者：
DAHLGREN, C