人外周血淋巴细胞化学发光的实验研究

EXPERIMENTAL STUDY ON THE CHEMILUMINESCENCEOF HUMAN PERIPHERAL BLOOD LYMPHOCYTES

目的研究各种有丝分裂原诱导的Ｌｙ－ＣＬ反应动力学特征和参与ＣＬ反应的细胞类型。方法从正常人外周血分离ＰＢＭＣ、淋巴细胞和Ｍｏ，经各种有丝分裂原诱导，测定和比较ＰＢＭＣ、淋巴细胞和Ｍｏ的ＣＬ反应强度及动力学。结果ＣｏｎＡ、ＰＨＡ、ＳＰＡ和ＰＷＭ均能诱导淋巴细胞产生ＣＬ反应，其最适刺激浓度分别为：ＣｏｎＡ１００μｇ／ｍｌ、ＰＨＡ１００μｇ／ｍｌ、ＳＰＡ１２５μｇ／ｍｌ、ＰＷＭ＞８００μｇ／ｍｌ。用平皿粘附法去除ＰＢＭＣ中的Ｍｏ，对Ｌｙ－ＣＬ反应无明显影响。Ｍｏ本身对ＣｏｎＡ只能产生缓慢、微弱的ＣＬ反应，而且其动力学与Ｌｙ－ＣＬ具有显著性差异。结论淋巴细胞是有丝分裂原诱导的ＰＢＭＣ－ＣＬ反应的主要产生细胞，Ｌｙ－ＣＬ测定是一种简便、快速检测淋巴细胞早期活化的有效方法。

Objective To investigate the kinetics of chemiluminescence(CL) and the cell types responsible for the generation of the CL in human peripheral blood lymphocytes induced by various mitogens. Methods Human peripheral blood mononuclear cells (PBMC),lymphocytes and monocytes (Mo) isolated from the peripheral blood of normal subjects were respectively stimuated with various mitogens. Results ConA, PHA, SPA and PWM were all capable of evoking the lymphocyte CL (Ly-CL) response.The maximal CL response was obtained after being stimulated with 100g/ml ConA,100g/ml PHA,125g/ml SPA and 800g/ml PWM respectively.LPS(625800g/ml) was unable to evoke Ly-CL response.Removal of adherent cells (Mo) from PBMC had no effect on Ly-CL.Monocytes could only generate very slow and weak CL response. There was significant difference between the CL kinetics of Mo and lymphocytes. Conclusion The results suggest that lymphocytes rather than monocytes are the main source of CL response induced by mitogens in human peripheral blood mononuclear cells.