Berneman Z N, van Bockstaele D R, Uyttenbroeck W M, Van Zaelen C, Cole-Dergent J, Muylle L, Peetermans M E
Labaratory of Experimental Hematology, University of Antwerp (UIA), Belgium.
Vox Sang. 1991;61(4):265-74. doi: 10.1111/j.1423-0410.1991.tb00958.x.
Blood group A antigen density on red blood cells (RBC) was studied using flow cytometry (FCM) and fluoresceinated polyclonal and monoclonal IgG anti-A antisera. Agglutination was a problem, which could only be solved by prefixation of the RBC with glutaraldehyde or formaldehyde. However, this treatment resulted in a significant reduction of the number of antigen sites as compared to the native (i.e. nonfixed) RBC. Two major new findings came out of this study: (1) A antigen density on native RBC seems to be higher than previously recognized, and (2) A antigen density distribution is probably non-Gaussian. The absolute number of A antigen sites was determined, using a human polyclonal IgG antiserum and commercially available absolute fluorescence standards. The site numbers on fixed RBC were comparable to those found by earlier radioimmunological studies (x 10(6)/RBC): A1, 1.07 +/- 0.28; A2, 0.21 +/- 0.09; A1B, 0.79 +/- 0.26 sites (mean +/- SD). The values found for native RBC were considerably higher (x 10(6)/RBC): A1, 2.86 +/- 0.95; A2, 0.47 +/- 0.29; A1B, 1.98 +/- 0.58 sites (mean +/- SD). With the 1 monoclonal and the 3 polyclonal antisera used in this study, and in contrast to Rh D, the erythrocytic A antigen density distribution of a given sample is highly asymmetrical. This non-Gaussian distribution profile does not seem to be affected by such factors as antibody heterogeneity, variability in antibody fluoresceination range, RBC density and reticulocyte content. This suggests that the asymmetrical A antigen distribution may be an intrinsic property of the RBC population.
使用流式细胞术（FCM）以及荧光标记的多克隆和单克隆IgG抗A抗血清，对红细胞（RBC）上的A血型抗原密度进行了研究。凝集是个问题，只有通过用戊二醛或甲醛对红细胞进行预固定才能解决。然而，与天然（即未固定）红细胞相比，这种处理导致抗原位点数量显著减少。这项研究得出了两个主要的新发现：（1）天然红细胞上的A抗原密度似乎比之前认为的要高，（2）A抗原密度分布可能不是高斯分布。使用人多克隆IgG抗血清和市售的绝对荧光标准品，测定了A抗原位点的绝对数量。固定红细胞上的位点数量与早期放射免疫研究的结果相当（×10⁶/RBC）：A1型，1.07±0.28；A2型，0.21±0.09；A1B型，0.79±0.26个位点（平均值±标准差）。天然红细胞的测定值要高得多（×10⁶/RBC）：A1型，2.86±0.95；A2型，0.47±0.29；A1B型，1.98±0.58个位点（平均值±标准差）。与Rh D不同，使用本研究中的1种单克隆和3种多克隆抗血清时，给定样本的红细胞A抗原密度分布高度不对称。这种非高斯分布模式似乎不受抗体异质性、抗体荧光标记范围的变异性、红细胞密度和网织红细胞含量等因素的影响。这表明A抗原的不对称分布可能是红细胞群体的固有特性。
