Application of immortalized human erythroid progenitor cell line in serologic tests to detect red blood cell alloantibodies

Transfusion. 2018 Nov;58(11):2675-2682. doi: 10.1111/trf.14840. Epub 2018 Sep 4.

Abstract

Background: 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.

Study design and methods: 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: 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.

Conclusion: 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.

MeSH terms

  • Anion Exchange Protein 1, Erythrocyte / genetics
  • Cell Line
  • Erythrocytes / immunology*
  • Erythroid Precursor Cells / cytology*
  • Erythroid Precursor Cells / immunology*
  • Flow Cytometry
  • Humans
  • Isoantibodies / immunology*
  • K562 Cells

Substances

  • Anion Exchange Protein 1, Erythrocyte
  • Isoantibodies
  • SLC4A1 protein, human