Department of Biochemistry & Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA.
Transfusion. 2011 May;51(5):1047-57. doi: 10.1111/j.1537-2995.2010.02926.x. Epub 2010 Nov 9.
Safe, accurate methods permitting simultaneous and/or repeated measurement of red blood cell (RBC) survival (RCS) are important to investigate pathophysiology and therapy of anemia. Methods using chromium 51 ((51) Cr)-labeled RBCs are unacceptable for infants, children, and pregnant women. We report RCS measured in vivo using RBCs labeled with several densities of biotin (BioRBCs).
Aliquots of autologous RBCs from eight healthy adult subjects were labeled separately at four discrete biotin densities, mixed, and infused. The proportion of each population of BioRBCs circulating was determined serially by flow cytometry over 20 weeks. For each population, RCS was assessed by the following: 1) posttransfusion BioRBC recovery at 24 hours (PTR(24) ); 2) time to decrease to 50% of the enrichment at 24 hours (T(50) ); and 3) mean potential lifespan (MPL).
Among the four BioRBC densities, no significant differences in PTR(24) were observed. T(50) and MPL were similar for the two lowest BioRBC densities. In contrast, the two highest BioRBC densities demonstrated progressively decreased T(50) and MPL.
RBCs labeled at four biotin densities can be used to independently and accurately measure PTR(24 ) and two lowest biotin densities can accurately quantitate long-term RCS. This method provides a tool for investigating anemia in infants, fetuses, and pregnant women with the following advantages over the standard (51) Cr method: 1) study subjects are not exposed to radiation; 2) small blood volumes (e.g., 20 µL) are required; and 3) multiple independent RCS measurements can be made simultaneously in the same individual.
安全、准确的方法可以同时或重复测量红细胞(RBC)的存活率(RCS),这对于研究贫血的病理生理学和治疗方法非常重要。使用铬 51(51)Cr 标记 RBC 的方法对于婴儿、儿童和孕妇是不可接受的。我们报告了使用几种生物素密度(BioRBC)标记的 RBC 进行体内测量的 RBC 存活率。
从 8 位健康成年受试者中分别取等量的自体 RBC,在四个不同的生物素密度下进行单独标记,混合并输注。通过流式细胞术在 20 周内连续测定循环中每个 BioRBC 群体的比例。对于每个群体,通过以下方法评估 RBC 存活率:1)输注后 24 小时的 BioRBC 恢复(PTR(24));2)24 小时内降低到富集量的 50%的时间(T(50));和 3)平均潜在寿命(MPL)。
在四种 BioRBC 密度中,PTR(24)没有明显差异。两种最低密度的 BioRBC 的 T(50)和 MPL 相似。相比之下,两种最高密度的 BioRBC 显示出逐渐降低的 T(50)和 MPL。
可以使用标记有四种生物素密度的 RBC 独立且准确地测量 PTR(24),并且两种最低生物素密度可以准确地定量长期 RBC 存活率。与标准的 51)Cr 方法相比,这种方法为研究婴儿、胎儿和孕妇的贫血提供了一种工具,具有以下优点:1)研究对象不会暴露于辐射下;2)需要的血量少(例如,20µL);3)可以在同一个体中同时进行多个独立的 RBC 存活率测量。