通过生物素标记,可对小鼠体内储存的人类血小板进行输血后功能评估。

IF 2.5 3区 医学 Q2 HEMATOLOGY
Transfusion Pub Date : 2024-07-01 Epub Date: 2024-05-17 DOI:10.1111/trf.17879
S Lawrence Bailey, Martin Bochenek, Aastha Chauhan, Brandon Miller, Moritz Stolla
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引用次数: 0

摘要

背景:使用放射性同位素对血小板进行放射性标记目前用于人体血小板恢复和存活研究。生物素化使体内外输血后血小板功能测试成为可能。研究设计和方法:研究设计:健康人的血小板浓缩物储存 6 天。研究设计:将健康人的血小板浓缩液储存 6 天,在 1 天、2 天和 6 天时采集样本,并按照放射性标记方案用生理盐水代替放射性铟-111 对血小板进行标记(假放射性标记 [sham-RL])。或者使用新开发的生物素化方案、洗涤方案或未经处理的对照样本。使用血小板激动剂刺激血小板并用血小板活化标记物标记血小板后,通过流式细胞术评估血小板功能。为了检测血小板是否能在输血后被激活,将标记的血小板输给非肥胖糖尿病/重症联合免疫缺陷小鼠,并在输血后 1 小时采集样本:结果:生物素化血小板的活化情况与输血前的假RL血小板相似,只是在储存第1/2天时α-脱颗粒明显减少,磷脂酰丝氨酸暴露增加。在储存第 6 天,假-RL 和生物素化血小板没有明显差异。假-RL 和生物素化血小板的活化性明显低于水洗血小板和未处理的对照血小板。输血后,生物素化血小板的活化情况与未处理的血小板基本无异:讨论:我们和其他人观察到的生物素化血小板活化水平的下降似乎主要是由于标记过程中的物理操作所致。总之,生物素化血小板可用于输血后功能评估,这是与放射性标记法相比的一大优势。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Biotin labeling allows for post-transfusion functional assessment of stored human platelets in mice.

Background: Platelet radiolabeling with radioisotopes is currently used for human platelet recovery and survival studies. Biotinylation enables ex vivo post-transfusion platelet function testing. Whether platelet biotinylation itself affects platelet function is controversial.

Study design and methods: Platelet concentrates from healthy humans were stored for 6 days. Samples were obtained at 1 or 2 and 6 days, and platelets were labeled following a radiolabeling protocol using saline instead of radioactive indium-111 (sham radiolabeling [sham-RL]). Alternatively, a newly developed biotinylation protocol, a washing protocol, or an unmanipulated control sample were used. Platelet function was assessed by flow cytometry after stimulation with platelet agonists and labeling of platelets with platelet activation markers. To test whether platelets can be activated after transfusion, labeled platelets were transfused into nonobese diabetic/severe combined immunodeficiency mice, and samples were obtained 1 h after transfusion.

Results: The activation profile of biotinylated platelets was comparable to sham-RL platelets before transfusion except for significantly less α-degranulation and more phosphatidyl serine exposure on storage day 1/2. There was no significant difference between sham-RL and biotinylated platelets on storage day 6. Sham-RL and biotinylated platelets were significantly less activatable than washed and unmanipulated control platelets. After transfusion, the activation profile of biotinylated platelets was largely indistinguishable from unmanipulated ones.

Discussion: The decrease in activation level in biotinylated platelets we and others observed appears mainly due to the physical manipulation during the labeling process. In conclusion, biotinylated platelets allow for post-transfusion function assessment, a major advantage over radiolabeling.

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来源期刊
Transfusion
Transfusion 医学-血液学
CiteScore
4.70
自引率
20.70%
发文量
426
审稿时长
1 months
期刊介绍: TRANSFUSION is the foremost publication in the world for new information regarding transfusion medicine. Written by and for members of AABB and other health-care workers, TRANSFUSION reports on the latest technical advances, discusses opposing viewpoints regarding controversial issues, and presents key conference proceedings. In addition to blood banking and transfusion medicine topics, TRANSFUSION presents submissions concerning patient blood management, tissue transplantation and hematopoietic, cellular, and gene therapies.
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