Premium
Identification of a novel DI*02(2558T) allele associated with weakened expression of DI2 antigen
Author(s) -
Wen Jizhi,
Akker Emile,
Luo Guangping,
Jia Shuangshuang,
Wei Ling,
Wang Zhen,
Schoot C. Ellen,
Ji Yanli
Publication year - 2020
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.16013
Subject(s) - allele , transfection , antigen , mutant , flow cytometry , western blot , sanger sequencing , phenotype , microbiology and biotechnology , biology , hek 293 cells , genetics , mutation , gene
Background The distribution of DI1/DI2 antigens of the Diego blood group system is polymorphic in Mongoloid populations and the corresponding alloantibodies are clinically significant. Here a novel DI variant was found by donor screening, and the effect of the novel and previously reported mutations on expression of DI1/DI2 antigens and Band 3 protein was explored. Study Design and Methods DNA samples of 1150 Chinese donors were collected. DI*01 / DI*02 genotyping was determined by Sanger sequencing. For the carrier of novel allele, the expression of Band 3 and DI1/DI2 antigens on red blood cells (RBCs) was detected by Western blot and flow cytometry, respectively. in vitro expression studies were conducted by transfecting the mutant (including the novel and three reported DI*02(2534T) , DI*02(2358_2359insCAC), and DI*02(2572T) alleles) or wild‐type DI*02 constructs into HEK 293T cells, the expression of Band 3 and DI1/DI2 antigens was analyzed. Results A novel heterozygous mutation (c.2558C>T, p.Thr853Met), which is located near the DI1/DI2 polymorphism site (c.2561T>C), was identified in a donor with DI:‐1,2 phenotype. Reduced expression of DI2 antigen was observed on the RBCs, while weakened expression of Band 3 and absence of DI2 antigen were detected in cells transfected with the mutant DI*02(2558T) construct. In addition, absent or decreased expression of Band 3 and DI2 antigen was also detected in cells transfected with three reported mutant constructs. Conclusion The novel DI*02(2558T) allele and three previously described DI mutations can affect the expression of Band 3 protein and/or DI2 antigen and/or interfere with DI*01/DI*02 genotyping result.