【摘要】 目的建立定量测定红细胞膜CD58分子的实验方法,研究其数量变化与肝硬化病情及免疫功能变化之间的关系。方法将戊二醛固定的1.2×106红细胞定量“液相包被”于V型板中,依次加入CD58分子单克隆抗体、碱性磷酸酶标记的第二抗体及可溶性底物,移显色液于比色孔,在405nm比色,计算其吸光值(A405)。结果本方法具有良好的敏感性和重复性。代偿性肝硬化患者红细胞CD58分子数量表达与正常人群无统计学差异。而失代偿性肝硬化患者的红细胞CD58数量表达,明显低于代偿性肝硬化患者和正常人群(P<0.01)。红细胞CD58分子表达变化,对红细胞调节淋巴细胞免疫黏附能力有明显影响。结论失代偿肝硬化红细胞CD58分子数量表达显著下降;红细胞CD58分子参与机体整体免疫调节。更多还原

【Abstract】 Objective To develop a technique to assay the expression of CD58 on erythrocytes and investigate the relationship between the quantity of CD58 expression and the severity and immune function in patients with hepatocirrhosis .Methods Glutaraldehyde-fixed red blood cells (RBC) were analyzed in V well microtitre plates. The primary antibodies of mouse monoclonal anti-human CD58 were added to the cells in each well. The microtitre plates were incubated at 37℃ for 45 min, followed by three washes in 1% S/BSA.Then the secondary antibodies of anti-mouse IgG conjugated to alkaline phosphatase were added, and was incubated in the same condition. Substrate was added and incubated at 37℃ for 90 min.The supernatant was transferred to a clean U microtitre plate to facilitate reading in a plate reader at 405 nm (A_ 405).Results The technique can nicely measure the quantity of CD58 expressed on erythrocytes, and has a good sensitivity and reproducibility. The A_ 405 of CD58 on erythrocytes in patients with compensated liver cirrhosis was not significantly different from that of healthy individuals, whereas the A_ 405 of CD58 on erythrocytes in patients with decompensated liver cirrhosis was significantly lower than those of patients with compensated liver cirrhosis and healthy individuals(P<0.01).The changes of CD58 expressed on erythrocytes can significantly influence the capability of erythrocytes in reglating the immune adherence of lymphocytes. Conclusions The quantity of CD58 expression on erythrocytes is significantly decreased in patients with decompensated liver cirrhosis. Quantitative expression of CD58 on erythrocytes plays an important role in the immune function of body.更多还原