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目的通过检测不同老化程度红细胞表面CD55、CD35和CD59数量,来探索老化红细胞清除机制中补体调节蛋白所发挥的作用。方法利用流式细胞术,检测30份标本其毛细管分离的年轻、年老红细胞表面CD35数量,和21份样本年轻、年老红细胞表面CD55和CD59数量。另外检测了其中随机选取的3份标本的4℃保存红细胞和37℃保存红细胞表面CD35、CD55和CD59数量。通过流式细胞术检测了15份标本自身IgG抗体的敏感性。结果与毛细管分离的年轻红细胞相比,年老红细胞CD35的平均荧光强度(MFI)下降了约35%(P<10-22)。年轻、年老红细胞表面CD55和CD59数量也呈现显著性下降(CD55:下降约44.17%;CD59:下降约36.18%,P<10-12)。体外保存的红细胞结果有所不同。对于CD35,4℃和37℃保存的红细胞表面数量变化不大无显著性统计学意义(P>0.05)。对于CD55和CD59,37℃保存的红细胞表面数量显著降低(P<0.01),而4℃保存的红细胞数量无显著性差异(P>0.05)。另外,毛细管分离得到的年老红细胞结合的自身抗体IgG显著高于年轻红细胞,荧光强度是年轻红细胞的1.8倍(P<0.01)。结论年轻、年老红细胞自身IgG抗体敏感性不同,说明毛细管离心方法可以一定程度反映人体生理条件下红细胞的老化状态。随着红细胞在血液循环中的老化,3种补体调节蛋白的显著下降,可能会使更多的C3b在红细胞表面沉积,从而中和调理、增强吞噬。此外,体外红细胞的老化与生理条件下的老化可能存在不同机制。
Objective To explore the role of complement regulatory proteins in the mechanism of erythrocyte clearance by detecting the numbers of CD55, CD35 and CD59 on the surface of erythrocytes with different degrees of aging. Methods Flow cytometry was used to detect the number of CD35 on the surface of young and old erythrocytes isolated from 30 samples and the numbers of CD55 and CD59 on the surface of young and old erythrocytes in 21 samples. In addition, the numbers of CD35, CD55 and CD59 on the surface of preserved erythrocytes at 4 ℃ and preserved erythrocytes at 37 ℃ were tested in 3 randomly chosen samples. Fifteen specimens were tested for their own IgG antibody sensitivity by flow cytometry. Results The mean fluorescence intensity (MFI) of CD35 in aged erythrocytes was decreased by about 35% (P <10-22) compared to capillary isolated red blood cells. The numbers of CD55 and CD59 on young and old erythrocytes also showed a significant decrease (CD55: a decrease of 44.17%; CD59: a decrease of 36.18%, P <10-12). Erythrocytes preserved in vitro have different results. There was no significant difference in the number of erythrocytes preserved at 4, 37 and 37 ℃ (P> 0.05). For CD55 and CD59, the number of erythrocytes preserved at 37 ℃ was significantly decreased (P <0.01), while there was no significant difference in the number of erythrocytes preserved at 4 ℃ (P> 0.05). In addition, IgG isolated from capillaries was significantly higher than that of young erythrocytes, and the intensity of fluorescence was 1.8 times that of young erythrocytes (P <0.01). Conclusion The sensitivities of young and old red blood cells to their own IgG antibodies are different, indicating that the capillary centrifugation method can reflect the aging status of red blood cells to a certain extent under physiological conditions. With the aging of red blood cells in the blood circulation, the significant decrease of 3 kinds of complement regulatory proteins may cause more C3b to deposit on the surface of erythrocytes, thereby neutralizing and enhancing phagocytosis. In addition, there may be different mechanisms of aging of erythrocytes in vitro and aging under physiological conditions.