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为了研究脐血淋巴细胞能否在体外培养成为特异性杀伤白血病细胞的杀伤性T细胞(CTL),联合细胞因子体外诱导脐血单个核细胞分化为树突状细胞(DC),再吞噬凋亡白血病细胞并将其抗原呈递给相同脐血的T淋巴细胞,得到杀伤性T细胞。用形态学及流式细胞术检测DC。CTL细胞的杀伤功能用乳酸脱氢酶(LDH)释放法测定。结果表明:12份脐血标本均可培养出形态典型的DC,DC的表面标志CD1a+、HLA DR+、CD86+、CD83+表达水平显著升高(P<0.05)。CTL可以杀伤未经培养的白血病细胞(效∶靶=50∶1对AML细胞的平均杀伤率为44.76±17.42%,对ALL细胞的平均杀伤率为8.50±4.25%),对相同患者缓解期的骨髓细胞杀伤率极低。结论: 在外体应用多种细胞因子刺激可诱导脐血单个核细胞分化成典型的DC;负载有白血病抗原的DC可诱导同一脐血的淋巴细胞生成白血病特异的杀伤性T细胞(CTL),所得CTL可特异性杀伤未经培养的白血病细胞而不严重伤害相同患者缓解期的骨髓细胞。
In order to investigate whether cord blood lymphocytes can be cultured in vitro as a specific cytotoxic killer T cell (CTL), combined with cytokines induced in vitro differentiation of cord blood mononuclear cells into dendritic cells (DC), and then engulf apoptosis Leukemia cells and their antigens presented to the same cord blood T lymphocytes, get killer T cells. DCs were examined by morphology and flow cytometry. The cytotoxicity of CTL cells was determined by lactate dehydrogenase (LDH) release assay. The results showed that DCs were cultured in all the 12 cord blood samples. The expression of CD1a +, HLA DR +, CD86 + and CD83 + on the surface of DCs was significantly increased (P <0.05). CTL can kill non-cultured leukemia cells (effect: the target = 50: 1 average killing rate of AML cells was 44.76 ± 17.42%, the average killing rate of ALL cells was 8.50 ± 4.25%), the same patient remission Bone marrow cell killing rate is very low. CONCLUSION: DCs can induce cord blood mononuclear cells to differentiate into typical DC in vitro by using a variety of cytokines. DCs loaded with leukemia antigen can induce lymphocyte-specific leukemia-specific killer T cells (CTL) from the same cord blood CTLs can specifically kill untreated leukemic cells without seriously injuring the bone marrow cells of the same patient during remission.