摘要
目的 建立一种定量测定红细胞CR1分子的敏感且重复性良好的实验方法 ,研究其临床价值。方法 戊二醛固定红细胞 ,将 3× 10 6的红细胞定量“液相包被”于V型板中 ,依次加入抗红细胞CR1的单抗、碱性磷酸酶标记的第二抗体及可溶性底物显色 ,移显色液于比色孔 ,在 40 5nm比色 ,计算其吸光度A值。结果 本方法具有良好的敏感性和重复性 ,正常健康人红细胞CR1分子数量表达显著高于肝细胞癌 (HCC)、大肠癌 (LIC)、风湿性心脏病 (RC)、卵巢癌 (OC)及系统性红斑狼疮(SLE)等病人 (P <0 0 1) ;SLE患者CR1分子的数量表达显著低于其他疾病 (P <0 0 1)。结论 本方法能准确测定出红细胞CR1分子表达的数量 ;红细胞CR1分子的数量表达及其变化与免疫相关性疾病特别是自身免疫性疾病的发生、发展与转归具有明显的相关性 ;定量测定红细胞CR1分子 ,对免疫相关性疾病的辅助诊断与治疗效果的评价具有重要的理论和临床价值。
Objective To investigate the role of CR1 on erythrocytes(ECR1) in immune-related diseases, we have developed a highly sensitive and reproductive technique to the examine number of the complement receptor 1 on erythrocytes (ECR1). Methods Glutaraldehyde-fixed red blood cells (RBC) were analysed in V well microtiter plates. To the cell buttons in each well, mouse monoclonal anti-CR1 was added and the microtiter was incubated at 37℃ for 50 min, followed by three washes in S/BSA. The anti-mouse IgG, conjugated to alkaline phosphatase was then 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 microtiter plate to facilitate reading in a plate reader at 405nm ( A ). Analysis the relationship between the A and the diseases. Results The method can nicely examine the quantity of CR1 on RBC and has a good sensitivity and reproducibility in methodology. The results showed that the ECR1 expression on RBC from patients with hepatocellular cancer (HCC), large intestine cancer (LIC), rheumatic cardiopathy (RC), ovary cancer(OC) and systemic lupus erythematosus (SLE) was significantly lower than that of healthy individuals( P <0 01). In particular, ECR1 on RBC in patients with SLE is significantly lower than that of patients with other diseases ( P <0 01). Furthermore, the CR1 expressed on RBC from the patients of SLE is obviously related to the development of the disease and the efficiency of treatment. Conclusions The technique has enabled us to investigate the role of ECR1 in immune-related diseases. The results suggest that the expression of ECR1 may be involved in pathogeny, development and prognosis of the diseases. It is very important to analyse the quantity of ECR1 in diagnosis and the evaluation of treatment of diseases.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2000年第4期381-384,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金!资助项目 (39670 754)
