Application of immortalized human erythroid progenitor cell line in serologic tests to detect red blood cell alloantibodies
Application of immortalized human erythroid progenitor cell line in serologic tests to detect red blood cell alloantibodies
复制标题
DOI：
10.1111/trf.14840
复制
发表时间：
2018-11-01
期刊：
影响因子：
2.9
通讯作者：
Satake, Masahiro
中科院分区：
文献类型：
--
作者：
Kikuchi, Go;Kurita, Ryo;Satake, Masahiro
背景研究设计和方法输血前试验中的抗体筛查对于避免输血的严重并发症是必要的。尽管表达相关同种异体抗原的红细胞（RBC）已被用于血清学抗体筛选，但很少注意使用其中转导了血型抗原基因的细胞系作为抗体筛选的试剂RBC。研究了红系祖细胞系在血清学试验中的应用。采用基因分型和流式细胞术分析红系祖细胞血型抗原的表达。使用红系祖细胞进行包括血凝在内的血清学分析，以评价其抗体检测的灵敏度。通过慢病毒转导进行外源性红系抗原的过表达，并研究抗体检测灵敏度。结果结论红系祖细胞含有大量的血红蛋白，并表达足够水平的血型抗原，以检测相应的单克隆抗体。此外，该细胞系可以获得一个外源性红细胞抗原慢病毒转导后，并检测相应的单克隆抗体和同种抗体具有相同的敏感性抗原阳性红细胞。应用红系祖细胞系筛选非预期抗体可能有助于解决与常规基于RBC的测定相关的试剂可用性等问题。通过基因修饰技术的红系祖细胞系的遗传扩增性可能导致更方便的试剂RBC的开发。
BACKGROUND STUDY DESIGN AND METHODS Antibody screening in pretransfusion tests is necessary to avoid critical complications of blood transfusion. Although red blood cells (RBCs) expressing relevant alloantigen(s) have been used for serologic antibody screening, little attention has been given to the use of cell lines, in which blood group antigen gene(s) are transduced, as reagent RBCs for antibody screening. The use of an erythroid progenitor cell line for serologic tests was studied. The expression of blood group antigens of erythroid progenitor cells was analyzed by genotyping and flow cytometry. Serologic analysis including hemagglutination was performed using erythroid progenitor cells to evaluate their sensitivity for antibody detection. Overexpression of exogenous erythroid antigen by lentiviral transduction was carried out and investigated for antibody detection sensitivity. RESULTS CONCLUSION Erythroid progenitor cells contained a substantial amount of hemoglobin and expressed sufficient levels of blood group antigens to detect corresponding monoclonal antibodies. Furthermore, the cell line could acquire an exogenous RBC antigen after lentiviral transduction and detected corresponding monoclonal and alloantibodies with equal sensitivity to antigen-positive RBCs. Application of erythroid progenitor cell lines for screening for unexpected antibodies could be helpful in solving issues such as reagent availability associated with the conventional RBC-based assay. The genetic expandability of erythroid progenitor cell lines by gene modification techniques could lead to the development of more convenient reagent RBCs.