淋巴细胞和巨噬细胞去除对人骨髓造血细胞体外扩增效应的影响

为了探讨淋巴细胞和巨噬细胞去除对人骨髓造血细胞体外扩增效应的影响 ,采用抗CD3、抗CD5 6和抗CD14单克隆抗体介导的补体细胞毒方法 ,去除骨髓单个核细胞 (MNC )中的T淋巴细胞、NK细胞和巨噬细胞 ,流式细胞技术 (FACS )分析细胞表面标志。将经不同单抗处理的骨髓MNC加入含有多种造血生长因子的培养基中进行体外有核细胞扩增 ,并观察多向祖细胞集落 (CFU GEMM )的形成能力。结果显示 ,经抗CD3、抗CD5 6、抗CD14单抗分别处理和用上述三种单抗共同处理的骨髓MNC中相应的靶细胞与MNC对照组相比均有明显的降低。经多种造血生长因子刺激培养 2 0d ,骨髓MNC对照组有核细胞总数扩增了 6 5倍 ,T细胞、NK细胞和巨噬细胞去除组分别扩增了 90倍、 77 2倍和 6 7 4倍 ,而三种细胞共同去除组仅为 2 86倍。骨髓MNC对照组CFU GEMM产率为 16 1 5 0± 12 5 4 ,T细胞去除组CFU GEMM产率为 370 0 0± 5 8 2 2 ,明显高于前者 ,差异显著 (P <0 0 0 5 )。NK细胞去除组为 2 0 5 5 0± 30 73,与MNC对照组相比也有显著差异 (P <0 0 5 )。巨噬细胞去除组为 16 3 2 5± 18 5 4 ,三种细胞共同去除组仅为 4 9 75± 9 98。本文结果提示 ,单纯去除骨髓MNC中的T淋巴细胞或NK细胞可明显促进人骨髓造血细胞的体外扩增效应。

Influences of Elimination of Lymphocytes and Macrophages on the Expansion of Human Bone Marrow Hematopoietic Cells in vitro

To investigate the effect of elimination of lymphocytes and macrophages on the expansion of bone marrow hematopoietic cells in vitro, T lymphocytes, NK cells and macrophages in human mononucleated cells were eliminated by means of complement-dependent cytotoxicity using the anti-CD3, anti-CD56 and anti-CD14 monoclonal antibodies, and the cell surface markers were analysed by FACS. After treatment with different monoclonal antibodies, bone marrow mononucleated cells were expanded in a liquid culture system with certain hematopoietic growth factors, and the number of cells in CFU-GEMM was determined. The results showed that in comparison with the untreated group, the numbers of target cells in the bone marrow mononucleated cells treated with anti-CD3, anti-CD56 and anti CD14 monoclonal antibodies or combined uses of these monoclonal antibodies were significantly decreased. After cocultivation with the hematopoitic growth factors for 20 days, the total numbers of the mononucleated cells expanded in the control cell group, and the T-cell, NK cell and macrophage-eliminated groups were 90, 77.2 and 67.4 folds respectively, but that with the 3 cells elimination was only 2.86 folds. The yields of CFU-GEMM in T cell-eliminated group, in NK cell eliminated group and in the macrophage-eliminated group were 370.00±58.22,205.50±30.73,and 163.25±18.54 respectively, while that in the 3 cells eliminated group was 49.75±9.98. These results indicate that elimination of T lymphocytes and NK cells from bone marrow can promote the expansion of human bone marrow hematopoitic cells in vitro.