测定同种异体反应中分泌细胞因子的T淋巴细胞及其临床意义初探

本研究目的是测定同种异基因外周血单个核细胞 (PBMNCs)刺激后分泌不同细胞因子的T淋巴细胞水平并对其临床意义进行研究 ,以建立一种监测同种异体反应性T淋巴细胞的新方法。首先 ,利用新颖的细胞因子分泌检测方法 (CKSA)从单细胞水平定量测定了人混合淋巴细胞反应后分泌IFN γ ,IL 4和IL 10的T淋巴细胞水平 ;然后根据上述实验结果 ,分析 2例接受异基因骨髓移植后发生急性移植物抗宿主病 (aGVHD)的患者外周血中分泌IFN γ的T细胞水平。结果表明 :经同种异体PBMNCs刺激后分泌IFN γ的T淋巴细胞水平显著升高 [(1.12± 0 .13) % ],而分泌IL 4和IL 10的T淋巴细胞则无升高 ,分别为 (0 .12± 0 .0 3) %和 (0 .10± 0 .0 3) %。患者外周血分泌IFN γ的T淋巴细胞水平和aGVHD的发生及严重程度有一定的相关性。结论 :利用CKSA从单细胞水平定量测定同种异体PBMNCs刺激后分泌IFN γ的T淋巴细胞水平的技术具有临床应用价值 ,可以应用于对aGVHD的识别。

Detection of Cytokine-Secreting T Lymphocytes in Allogeneic Reaction and A Preliminary Study on Its Clinical Significance

In order to explore a new way to study allogeneic reactive T lymphocytes, detection of cytokine-secreting T lymphocytes after allogeneic peripheral blood mononuclear cells (PBMNCs) stimulation and investigation of its clinical significance were performed. A novel cytokine secretion assay (CKSA) was first applied to detect T lymphocytes secreting cytokine including IFN-gamma, IL-4 and IL-10 at single cell level in human mixed lymphocytes reaction. IFN-gamma-secreting T cells from PBMNCs were then evaluated in 2 patients with acute graft versus host disease (aGVHD) after allogeneic bone marrow transplantation. The results showed that compared with IL-4 and IL-10 (which were 0.12 +/- 0.03% and 0.10 +/- 0.03% respectively), a sizable proportion of IFN-gamma-secreting T lymphocytes could be detected (1.12 +/- 0.13)% after allogeneic PBMNCs stimulation. Preliminary results indicated that frequency of IFN-gamma-secreting T lymphocytes correlated with the onset and severity of clinical aGVHD. In conclusion, it is feasible to detect IFN-gamma secreting T lymphocytes after allogeneic PBMNCs stimulation and to apply the CKSA technique for clinical identification of aGVHD.