Identification of Stages of Erythroid Differentiation in Bone Marrow and Erythrocyte Subpopulations in Blood Circulation that Are Preferentially Lost in Autoimmune Hemolytic Anemia in Mouse
Identification of Stages of Erythroid Differentiation in Bone Marrow and Erythrocyte Subpopulations in Blood Circulation that Are Preferentially Lost in Autoimmune Hemolytic Anemia in Mouse
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DOI：
10.1371/journal.pone.0166878
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发表时间：
2016-11-21
期刊：
影响因子：
3.7
通讯作者：
Saxena, Rajiv K.
中科院分区：
文献类型：
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作者：
Chatterjee, Sreoshi;Bhardwaj, Nitin;Saxena, Rajiv K.
每周重复注射大鼠红细胞，5-6 周后，C57BL/6 小鼠产生自身免疫性溶血性贫血 (AIHA)。使用我们实验室最近开发的双体内生物素化 (DIB) 技术，监测 AIHA 期间不同年龄组红细胞群的周转情况。结果显示，血液循环中网织红细胞、中青年红细胞比例明显下降，老年红细胞比例明显上升。自身抗体与年轻红细胞的结合相对较高，并且在这些细胞中还观察到较高水平的细胞内活性氧（ROS）。通过监测这些器官中处于不同分化阶段的红细胞的相对比例来检查AIHA诱导小鼠的骨髓和脾脏中的红细胞生成活性。通过使用抗 Ter119 和抗转铁蛋白受体 (CD71) 单克隆抗体双染色，对不同分化阶段的细胞进行流式细胞术计数。在 AIHA 诱导的小鼠中，骨髓中的红细胞明显下降，其中成红细胞 C 受影响最大（下降 50%）。成红细胞 C 还记录了高细胞内 ROS 水平以及膜结合自身抗体水平的增加。脾脏中没有观察到这种下降。已提出的 AIHA 模型表明自身抗体的结合可能不是破坏骨髓和血液循环中红细胞的充分条件。骨髓中红细胞生成分化的最后阶段和血液循环中红细胞的早期阶段特别容易在 AIHA 中被去除。
Repeated weekly injections of rat erythrocytes produced autoimmune hemolytic anemia (AIHA) in C57BL/6 mice after 5-6 weeks. Using the double in vivo biotinylation (DIB) technique, recently developed in our laboratory, turnover of erythrocyte cohorts of different age groups during AIHA was monitored. Results indicate a significant decline in the proportion of reticulocytes, young and intermediate age groups of erythrocytes, but a significant increase in the proportion of old erythrocytes in blood circulation. Binding of the autoantibody was relatively higher to the young erythrocytes and higher levels of intracellular reactive oxygen species (ROS) were also seen in these cells. Erythropoietic activity in the bone marrows and the spleen of AIHA induced mice was examined by monitoring the relative proportion of erythroid cells at various stages of differentiation in these organs. Cells at different stages of differentiation were enumerated flow cytometrically by double staining with anti-Ter119 and anti-transferrin receptor (CD71) monoclonal antibodies. Erythroid cells in bone marrow declined significantly in AIHA induced mice, erythroblast C being most affected (50% decline). Erythroblast C also recorded high intracellular ROS level along with increased levels of membrane-bound autoantibody. No such decline was observed in spleen. A model of AIHA has been proposed indicating that binding of autoantibodies may not be a sufficient condition for destruction of erythroid cells in bone marrow and in blood circulation. Last stage of erythropoietic differentiation in bone marrow and early stages of erythrocytes in blood circulation are specifically susceptible to removal in AIHA.