Purpose

Closed eye neutrophils have demonstrated increased prevalence in dry eye disease, but the phenotype and extent of activation of these cells has yet to be described.

Methods

12 normal subjects and 12 subjects with dry eye disease were recruited and trained for self-collection of closed eye leukocytes, immediately upon awakening. Tear leukocytes were isolated and peripheral blood was collected, and stained with a panel of fluorescently-labeled antibodies to determine the activation phenotype of neutrophils. Extracellular matrix metalloproteinase 9 (MMP9) and neutrophil elastase (NE) was quantified by an enzyme-linked immunosorbent assay.

Results

Total numbers of tear leukocytes recovered, at awakening, from normal and dry eye subjects were similar. Tear neutrophils from dry eye subjects had increased expression of membrane receptor CD66b, a marker associated with secondary granule degranulation. There was also a higher proportion of monocytes in the dry eye cohort, as compared to the normal cohort. Extracellular MMP9 was significantly higher in subjects with dry eye disease, and while NE was also elevated, it did not achieve statistical significance.

Conclusions

Increased inflammation can be observed in the closed eye tears of subjects with dry eye disease, and neutrophils may be a potential source of pathogenic species in dry eye disease. Further research is required to determine the diagnostic potential of closed eye tears.

中文翻译：

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在干眼病中，闭眼中性粒细胞脱粒增强。

目的

闭眼中性粒细胞已显示出在干眼病中的患病率增加，但是这些细胞的表型和活化程度尚未描述。

方法

觉醒后立即招募和训练12名正常受试者和12名患有干眼病的受试者进行闭眼白细胞自我收集。分离眼泪白细胞并收集外周血，并用一组荧光标记的抗体染色以确定嗜中性粒细胞的激活表型。细胞外基质金属蛋白酶9（MMP9）和嗜中性粒细胞弹性蛋白酶（NE）通过酶联免疫吸附测定进行定量。

结果

觉醒时从正常和干眼受试者中恢复的眼泪白细胞总数相似。来自干眼受试者的泪中性粒细胞具有增加的膜受体CD66b的表达，CD66b是与继发性颗粒脱粒相关的标志物。与正常人群相比，干眼人群中单核细胞的比例也更高。干眼病患者的细胞外MMP9明显更高，而NE也升高，但未达到统计学意义。

结论

在患有干眼病的受试者的闭合眼泪中可以观察到炎症的增加，并且中性粒细胞可能是干眼病中病原体的潜在来源。需要进一步的研究以确定闭合性眼泪的诊断潜力。