Flow cytometric analysis of the Rh1 (Rho, D) antigen activity on red cells: various Rh blood group phenotypes including du variants
Ota, M.; Hasekura, H.; Fukushima, H.; Yonemura, I.
Nihon Hoigaku Zasshi 43(2): 122-127
1989
ISSN/ISBN: 0047-1887 PMID: 2509769 Document Number: 334134
Rh1 (Rho, D) antigen activity has been analyzed by the use of the indirect immunofluorescence flow cytometry (FCM), and the Rh blood group genotypes were able to be successfully determined from the intensity of fluorescence detected in flow cytometry using the anti-D IgG that was fractionated in a Protein A Sepharose CL-4B column as the primary antibody. The relative amount of the fluorescein isothiocyanate (FITC) bound to the D (R1R1, CDe/CDe), the high grade Du (R2r',cDE/Cde), the low grade Du (K1r, CDue/cde), and the d (rr, cde/cde) red cells was estimated from the mean fluorescent intensity. The FITC-binding activity of the high grade Du and low grade Du was 83% and 21% that of D. The antigen-antibody complex density profile was analyzed by using the FITC-conjugated protein-A in place of the second antibody. Compared with the found results using anti-human globulin as the second antibody, this method was less sensitive but it still was able to demonstrate the different degrees of fluorescence according to the Rh genotypes. The present FCM method is both simple and useful for (1) measuring the relative amount of antigens, (2) for detecting the dosage effect and (3) for deferminins the blood group genotypes.