Lietaer Leen, Demeyere Kristel, Heirbaut Stijn, Meyer Evelyne, Opsomer Geert, Bogado Pascottini Osvaldo
Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium.
Department of Pharmacology, Toxicology and Biochemistry, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium.
Animals (Basel). 2021 Apr 10;11(4):1081. doi: 10.3390/ani11041081.
Postpartum dairy cows experience impaired peripheral polymorphonuclear leukocyte (PMN) functionality, which has been associated with reproductive tract inflammatory diseases. However, it has not been elucidated yet whether endometrial PMN functionality is (equally) impaired. We developed a method for endometrial PMN isolation and flow cytometric assessment of their viability and functionality. We also evaluated PMN immunolabeling, using a specific bovine granulocyte marker, CH138A. Blood and endometrial cytobrush samples were collected in duplicate from seventeen clinically healthy Holstein-Friesian cows between 9 and 37 days in milk. The proportion of viable, apoptotic, and necrotic PMN in endometrial samples roughly ranged from 10 to 80%, indicating highly dynamic endometrial PMN populations in the postpartum uteri. Endometrial PMN functionality testing revealed that PMN immunolabeling increased the accuracy, although this protocol might influence the median fluorescence intensity of the sample. Phagocytosis seemed the most stable and reliable endometrial PMN function and could be assessed satisfactorily without prior CH138A immunolabeling. However, the interpretation of oxidative burst and intracellular proteolysis tests remains challenging. The correlation between peripheral and endometrial PMN functionality was poor. Further research is warranted to unravel the role of uterine PMN viability and functionality in bovine uterine health.
产后奶牛的外周多形核白细胞（PMN）功能受损，这与生殖道炎症性疾病有关。然而，子宫内膜PMN功能是否（同样）受损尚未阐明。我们开发了一种用于分离子宫内膜PMN并通过流式细胞术评估其活力和功能的方法。我们还使用特异性牛粒细胞标志物CH138A评估了PMN免疫标记。从17头临床健康的荷斯坦 - 弗里生奶牛在产奶9至37天期间重复采集血液和子宫内膜细胞刷样本。子宫内膜样本中存活、凋亡和坏死PMN的比例大致在10%至80%之间，表明产后子宫内的子宫内膜PMN群体具有高度动态性。子宫内膜PMN功能测试表明，PMN免疫标记提高了准确性，尽管该方案可能会影响样本的中位荧光强度。吞噬作用似乎是最稳定可靠的子宫内膜PMN功能，无需事先进行CH138A免疫标记即可令人满意地评估。然而，氧化爆发和细胞内蛋白水解测试的解读仍然具有挑战性。外周和子宫内膜PMN功能之间的相关性较差。有必要进行进一步研究以阐明子宫PMN活力和功能在奶牛子宫健康中的作用。
