摘要
目的 :初步探讨钙离子载体 (calciumionophore,CI)诱导人外周血单个核细胞 (PBMC)向树突状细胞 (DC)分化的细胞信号转导途径。方法 :分离健康献血者的PBMC ,加入rhGM CSF及A2 3187各 10 0 μg/L ,部分细胞预先用W 7(10 μmol/L)或CsA(0 .5mg/L)或KT5 92 6 (1μmol/L)处理 30min后 ,再加入rhGM CSF及A2 3187各 10 0 μg/L。体外培养 4 0h后 ,于相差显微镜下观察细胞的形态 ;流式细胞仪检测细胞的表面标志 ;用MTT比色法检测上述细胞刺激同种异体T细胞的增殖作用。结果 :与 10 0 μg/L的A2 3187及rhGM CSF共同培养 4 0h的健康献血者的PBMC ,出现典型的树突状突起 ,同时CD83、CD80及CD86分子的表达上调 ,CD14分子的表达下调 ,刺激同种异体T细胞增殖的能力增强 ;而预先用W 7或CsA或KT5 92 6处理 30min后、再给予rhGM CSF及A2 3187处理的PBMC ,其形态、表面标志物及对T细胞的刺激增殖能力 ,均不同程度的受到抑制。结论 :CI诱导的PBMC向DC的分化 ,可能受控于Ca2 +
AIM: To explore the intracellular signal transduction pathway in the differentiation of human peripheral blood mononucleocytes (PBMCs) towards dendritic cells (DCs) induced by calcium ionophore (CI). METHODS: PBMCs isolated from a healthy donor were cultured with A23187 plus rhGM-CSF, 100 μg/L each. In some experiments, PBMCs were cultured for 30 minutes with W-7 (10 μmol/L), CsA(0.5 mg/L) or KT5926(1 μmol/L) before addition of rhGM-CSF and A23187. After culture for 40 hours, morphological change of the cells were observed under phase contrast microscope; surface markers on treated PBMCs were analyzed by flow cytometry; the proliferation of allogeneic human T cells stimulated by the treated PBMCs was detected by MTT colorimetry. RESULTS: PBMCs of the healthy donor cocultured with rhGM-CSF plus CI for 40 hours had the typical morphology of DCs, with decreased CD14 expression, and increased CD83, CD80 and CD86 expressions. The proliferation of allogeneic T cells stimulated by PBMCs treated with A23187 plus rhGM-CSF was strengthened. But the morphological changes, surface marker expressions and the ability to enhancing proliferation of allogeneic T cells were inhibited to different degrees by W-7, CsA or KT5926. CONCLUSION: The differentiation of PBMCs towards DCs by CI may be modulated by Ca 2+/calmodulin and multiple signal transduction pathways downstream.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2004年第5期540-543,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
广东省科技计划基金资助项目 (No .2 0 0 3A30 2 0 30 1 )
