外周血淋巴细胞亚群检测及其rDNA转录活性分析

DETECTION OF PERIPHERAL BLOOD LYMPHOCYTE SUBTYPE AND ANALYSIS OF THE rDNA TRANSCRIPTIONAL ACTIVITY

目的 分析培养后外周血淋巴细胞亚群的构成特征，检测淋巴细胞ｒＤＮＡ转录活性，探讨其在肿瘤发生及诊断中的意义。 方法 对２１ 例健康人及１２７ 例良恶性肿瘤患者的外周血培养７２ 小时，用标记ＣＤ４ 、ＣＤ８、ＣＤ２０、ＣＤ５６ 对培养后外周血作流式细胞术分析，同时用ＡｇＮＯＲ染色结合图像分析检测淋巴细胞的Ｉ－Ｓ％ （银染核仁积分面积细胞核积分面积） 及Ｉ－Ｏ－Ｄ％ （银染核仁积分光密度细胞核积分光密度） 以反映ｒＤＮＡ转录活性。 结果 外周血培养后主要为Ｔ淋巴细胞的转化生长，恶性肿瘤患者的Ｔ淋巴细胞ｒＤＮＡ 转录活性显著低于健康人或良性患者（Ｐ＜０－０５），表明恶性肿瘤患者的细胞免疫功能较低。 结论 外周血Ｔ淋巴细胞ｒＤＮＡ转录活性的检测有助于恶性肿瘤的诊断与鉴别诊断，并可能有助于肿瘤的普查和筛选。用图像分析系统检测外周血淋巴细胞ｒＤＮＡ转录活性，快速、客观、分辨率高，并可作定量分析。

analyze the cultured peripheral blood lymphocyte(PBL) subtype and to detect the rDNA transcriptional activity,further to explore the significance in development and diagnosis of malignancies.METHODS Peripheral blood from 21 normal volunteers and 127 benign and malignant patients were cultured for 72 hours.Flow cytometry analysis was performed with the cultured PBL by labelled CD4,CD8,CD20 and CD56.The staining of AgNOR combined image analysis were used to detect the I.S% and I.O.D% which reflected the rDNA transcriptional activity.RESULTS After 72 hours cultured peripheral blood mainly showed T lymphocyte transformed growth.The rDNA transcriptional activities in malignant patients were lower than normal persons or benign patients ( P <0 05),showing the cellular immunity in malignant patients were lower.CONCLUSIONS The detection of the rDNA transcriptional activity of peripheral T lymphocyte will be helpful for diagnosis and differential diagnosis of malignancies,and may be helpful for the screening of malignancies.Image analysis for the detection of the rDNA transcriptional acitivity is rapid,objective,it has high resolving power and can perform quantitative analysis.It is a good method to be highly recommended.