| Research Abstract |
1. Rhl(Rho,D),HLA-A2,HLA-A24,HLA-Bw52 and HLA-DR2 antigen activities have been analyzed by the use of indirect FCM. Various RH blood group genotypes including rare variants expressing Del and D^u(high-grade and low-grade)were successfully determined from the intensity of fluorescence detected in FCM using anti-D IgG fractionated in a Protein A Sepharose CL-4B clolumn as the primary antibody. Also, the HLA genotypes could be successfully determined from the different degrees of fluorescence intensity detected by using FITC(fluorescein isothiocyanate) conjugated anti-human IgG goat F(ab)' serum as the secondary antibody.
2. After sensitization by anti-D serum, the ^<125>-protein A bindings were investigated in D, Del, and d red cells. The relative amount of ^<125>-protein A bound to Del cells was less than one per cent of the D cells.
3. Genomic DNAs from 12 normal individual cells expressing DR1,DR2,DRlDR2,DR2DRW8,DRlDRw8,DRw8,DR1DR9 and DR9 antigens were digested with 3 different restriction endonucleases(EcoRI,MspI and TagI) and hybridized to a DQbeta cDNA probe. Distribution analysis of restriction fragment length polymorphism (RFLP) was able to differentiate the zygosity of DR antigens(a DR homozygote or a DR heterozygote involving DR-gene).
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