Dircks Brigitte Hedwig, Mischke Reinhard, Schuberth Hans-Joachim
Small Animal Clinic, University of Veterinary Medicine Hanover, 30559 Hanover, Germany.
Am J Vet Res. 2012 Jul;73(7):939-45. doi: 10.2460/ajvr.73.7.939.
To evaluate platelet-neutrophil aggregate (PNA) formation and neutrophil shape as indicators of neutrophil activation in dogs with systemic inflammatory diseases and after blood sample incubation with various platelet and neutrophil agonists.
20 dogs with systemic inflammatory response syndrome (SIRS) and 10 healthy Beagles.
Neutrophils were isolated from blood samples directly after blood sample collection and after incubation of blood samples with phorbol myristate acetate, collagen, adenosine diphosphate, epinephrine, or various concentrations of lipopolysaccharide or arachidonic acid. CD61+ neutrophils as an indicator of PNA formation were evaluated, and neutrophil size and granularity were assessed via flow cytometry.
Dogs with SIRS had more PNA formation, larger neutrophil size, and less granularity relative to control dogs, but no differences were evident when these dogs were grouped by whether they had sepsis (n = 6) or disseminated intravascular coagulation (12). A significant increase in PNA formation occurred after neutrophil incubation with all agonists, and incubation with phorbol myristate acetate elicited the strongest response. Neutrophils increased in size and decreased in granularity after incubation with all agonists except epinephrine. Incubation with lipopolysaccharide or arachidonic acid resulted in a dose-dependent effect on PNA formation and neutrophil shape.
SIRS appeared to increase the degree of PNA formation and neutrophil shape change. Similar changes after neutrophil incubation with platelet agonists suggested that platelet activation has a role in PNA formation. Additional studies are necessary to determine the clinical importance and diagnostic value of PNA formation in dogs with SIRS and sepsis.
评估血小板 - 中性粒细胞聚集体（PNA）的形成以及中性粒细胞形态，以此作为患有全身性炎症疾病的犬以及血液样本与各种血小板和中性粒细胞激动剂孵育后中性粒细胞活化的指标。
20只患有全身性炎症反应综合征（SIRS）的犬和10只健康的比格犬。
在采集血样后直接从血样中分离中性粒细胞，并在血样与佛波酯肉豆蔻酸酯乙酸盐、胶原蛋白、二磷酸腺苷、肾上腺素或不同浓度的脂多糖或花生四烯酸孵育后分离。评估作为PNA形成指标的CD61 +中性粒细胞，并通过流式细胞术评估中性粒细胞大小和颗粒度。
与对照犬相比，患有SIRS的犬有更多的PNA形成、更大的中性粒细胞大小以及更低的颗粒度，但根据这些犬是否患有败血症（n = 6）或弥散性血管内凝血（12）进行分组时，差异并不明显。中性粒细胞与所有激动剂孵育后，PNA形成显著增加，与佛波酯肉豆蔻酸酯乙酸盐孵育引发的反应最强。除肾上腺素外，中性粒细胞与所有激动剂孵育后大小增加且颗粒度降低。与脂多糖或花生四烯酸孵育对PNA形成和中性粒细胞形态产生剂量依赖性影响。
SIRS似乎增加了PNA形成程度和中性粒细胞形态变化。中性粒细胞与血小板激动剂孵育后出现类似变化表明血小板活化在PNA形成中起作用。需要进一步研究以确定PNA形成在患有SIRS和败血症的犬中的临床重要性和诊断价值。
