Institute of Health Service and Transfusion Medicine, Academy of Military Medical Sciences, Beijing, China.
Transfusion. 2019 May;59(5):1799-1808. doi: 10.1111/trf.15183. Epub 2019 Feb 8.
Cryopreservation in dimethyl sulfoxide and storage at -80 °C extends the shelf life of platelets to at least 2 years, allowing greater availability in rural and military areas. While cryopreserved platelets (CPPs) have been extensively characterized for coagulation and thrombin generation, reports on the mechanism of adverse reactions to CPPs transfusion are scarce. Here, we tested the hypothesis that CPPs facilitate phagocytosis by Kupffer cells and subsequently promote the inflammatory response in Kupffer cells.
P-selectin expression, glycoprotein Ibα clustering and phosphatidylserine (PS) surface exposure on platelets stored at 22 °C, 4 °C and - 80 °C for 3 days were examined by flow cytometry. The phagocytosis of mepacrine-labeled platelets coincubated with THP-1 cells was examined by flow cytometry and confocal microscopy, and the release of cytokines from THP-1 cells was measured by enzyme-linked immunosorbent assay.
CPPs showed a marked enhancement of exposed PS but dramatically reduced glycoprotein Ibα expression and clustering compared with platelets stored at 4 °C. Activation of THP-1 cells was stronger by CPPs than by platelets stored at 22 °C and 4 °C. CPP interference tests using annexin V and anti-P-selectin showed that CPPs induced increases in PS- and P-selectin-mediated phagocytosis, as well as secretion of the proinflammatory cytokine tumor necrosis factor-α, and interleukins IL-1β and IL-6, but a decrease in transforming growth factor-β production in THP-1 cells. Surface-exposed PS was more effective than P-selectin for the activation of THP-1 cells.
CPPs triggered PS and P-selectin-mediated phagocytosis by macrophages and stimulated the inflammatory response of macrophages.
在二甲基亚砜中冷冻保存并储存在-80°C 下可将血小板的保质期延长至至少 2 年，从而在农村和军事地区增加其可用性。虽然已广泛研究了冷冻保存的血小板（CPP）在凝血和血栓生成方面的特性，但关于 CPP 输血不良反应机制的报告却很少。在这里，我们检验了以下假设：CPP 促进了枯否细胞的吞噬作用，随后促进了枯否细胞的炎症反应。
通过流式细胞术检查了在 22°C、4°C 和-80°C 下储存 3 天的血小板中 P-选择素表达、糖蛋白 Ibα 聚集和磷脂酰丝氨酸（PS）表面暴露情况。通过流式细胞术和共聚焦显微镜检查了与 THP-1 细胞共孵育的 Mepacrine 标记的血小板的吞噬作用，并通过酶联免疫吸附试验测量了来自 THP-1 细胞的细胞因子释放。
与在 4°C 下储存的血小板相比，CPP 显示出明显增强的 PS 暴露，但糖蛋白 Ibα表达和聚集却大大降低。CPP 对 THP-1 细胞的激活作用强于在 22°C 和 4°C 下储存的血小板。用 Annexin V 和抗 P-选择素进行 CPP 干扰试验表明，CPPs 诱导 PS 和 P-选择素介导的吞噬作用增加，以及促炎细胞因子肿瘤坏死因子-α、白细胞介素-1β和白细胞介素-6 的分泌增加，但转化生长因子-β的产生减少THP-1 细胞。表面暴露的 PS 比 P-选择素更有效地激活 THP-1 细胞。
CPPs 触发了巨噬细胞的 PS 和 P-选择素介导的吞噬作用，并刺激了巨噬细胞的炎症反应。
