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  • 1
    In: Transfusion, Wiley, Vol. 58, No. 11 ( 2018-11), p. 2675-2682
    Abstract: 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.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2018
    detail.hit.zdb_id: 2018415-3
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  • 2
    In: Transfusion, Wiley, Vol. 61, No. 10 ( 2021-10), p. 2825-2829
    Abstract: In this study, we identified a novel glycophorin variant (GP.MOT) in a Mi a ‐positive Japanese blood donor. The proband with this glycophorin variant was discovered by antigen screening of samples from 475,493 Japanese blood donors using monoclonal anti‐Mi a . Study design and methods Standard serological techniques and flow cytometry were performed. GP.MOT RBCs were examined by immunoblotting using anti‐GPA, anti‐MUT or anti‐Mur. Genome DNA was extracted from whole blood, and the GYPA / GYPB was analyzed by polymerase chain reactions and Sanger sequencing. Results The MNS blood group of the proband was M + N +  w S‐s + with the presence of other low‐frequency antigens including Mi a , Mur, MUT, and KIPP. A 43‐kDa molecule, which is almost equivalent in size to glycophorin A (GPA), was identified by immunoblotting using monoclonal anti‐MUT and anti‐Mur. Sanger sequencing clearly indicated that the proband had two different GYPA*M alleles at SNP rs62334651 ( GYPA*M232 + 55A and GYPA*M232 + 55G ), as well as a GYP ( B‐A ) hybrid allele ( GYP*MOT ) with breakpoints located on pseudoexon 3 of GYPB from c.210 to c.219. Discussion We identified a hybrid glycophorin GP.MOT with the deduced unique amino acid sequence GPB (20–45)‐GP Ψ B (46–70)‐GPA (71–149), which has not been previously reported.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2021
    detail.hit.zdb_id: 2018415-3
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  • 3
    In: Vox Sanguinis, Wiley, Vol. 114, No. 2 ( 2019-02), p. 171-173
    Abstract: We found an individual with weakened S antigen expression on red blood cells ( RBC s) during routine blood grouping. The proband was typed S+s+ by polyclonal antibodies, but the RBC s demonstrated different reactivity with three monoclonal anti‐S. The proband did not have alloanti‐S. Cloning and Sanger sequencing revealed that the proband had a c.166A 〉 T (p.Thr56Ser) mutation in exon 4 of GYPB * S . When antibody screening of 60 455 blood donors was performed using the proband RBC s, no antibodies were detected. GYPB *S with c.166T should encode an unusual S antigen but the creation of a novel antigen remains to be investigated.
    Type of Medium: Online Resource
    ISSN: 0042-9007 , 1423-0410
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2019
    detail.hit.zdb_id: 1483587-3
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  • 4
    In: Vox Sanguinis, Wiley, Vol. 115, No. 8 ( 2020-11), p. 756-766
    Abstract: The low‐incidence antigen St a of the MNS system is usually associated with the GP(B‐A) hybrid molecule, which carries the ‘N’ antigen at the N terminus. The GP(A‐A) molecule with trypsin‐resistant M antigen has been found in a few St(a+) individuals. Materials and Methods Among Japanese blood donors, we screened 24 292 individuals for the presence of St(a+) with trypsin‐resistant ‘N’ antigen and 193 009 individuals for the presence of St(a+) with trypsin‐resistant M antigen. The breakpoints responsible for the St a antigen were analysed by sequencing the genomic DNAs. Results A total of 1001 (4·1%) individuals were identified as St(a+) with trypsin‐resistant ‘N’ antigen. Out of 1001 individuals, 115 were selected randomly for sequencing. Two novel GYP*Sch ( GYP*401 ) variants with new intron 3 breakpoints of GYPA were detected in three cases. Twenty‐five (0·013%) individuals were identified as St(a+) with trypsin‐resistant M antigen. Five individuals had the GYP(A‐ψB‐A) hybrid allele; two of these five individuals were GYP*Zan ( GYP*101.01 ), and the remaining three had a novel GYP(A‐ψB‐A) allele with the first breakpoint in GYPA exon A3 between c.178 and c.203. Nine individuals had a novel GYP(A‐E‐A) allele with GYPE exon E2 and pseudoexon E3 instead of GYPA exon A2 and A3. The 11 remaining individuals had a novel GYP(A‐A) allele with a 9‐bp deletion that included the donor splice site of intron 3 of GYPA . Conclusion Our finding on diversity of glycophorin genes responsible for St a antigen provides evidence for further complexity in the MNS system.
    Type of Medium: Online Resource
    ISSN: 0042-9007 , 1423-0410
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2020
    detail.hit.zdb_id: 1483587-3
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