脐带血树突状细胞的生物活性和抗白血病作用

IF 0.6 Q4 MEDICINE, RESEARCH & EXPERIMENTAL International journal of clinical and experimental medicine Pub Date : 2015-10-15 eCollection Date: 2015-01-01
Xu-Cang Wei, Di-Di Yang, Xiu-Rui Han, Yu-An Zhao, Yan-Chun Li, Li-Jie Zhang, Jiu-Ju Wang
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摘要

研究目的我们研究了脐带血树突状细胞(DC)对体外增殖、免疫表型和同源细胞因子诱导的杀伤细胞(CIK)水平的影响以及对白血病细胞的毒性:方法:收集来自脐带血的单核细胞诱导的 DC-CIK 细胞,按 1:5 的比例共培养。脐带血 CIK 细胞或外周血 DC-CIK 细胞作为对照。用流式细胞仪分析表型;用胰蓝进行小瓶细胞计数;用 MTT 法测量效应细胞对白血病细胞的杀伤活性。用酶联免疫吸附法测定干扰素-r(IFN-r)、肿瘤坏死因子-a(TNF-α)和白细胞介素-12(IL-12)的水平:结果:与脐带血CIK细胞和外周血DC-CIK细胞(PC)相比,DC-CIK细胞的增殖能力明显提高:结论:脐带血DC可增强同源CIK细胞的增殖能力和抗白血病作用。虽然脐带血DC-CIK细胞的增殖能力高于外周血DC-CIK细胞,但两种DC-CIK细胞在细胞毒性方面并无明显差异。脐带血DC-CIK细胞的可用性高、移植排斥反应的概率低,在免疫疗法中的应用前景更为广阔。
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Bioactivity of umbilical cord blood dendritic cells and anti-leukemia effect.

Objective: We investigated the effect of umbilical cord blood dendritic cells (DCs) on in vitro proliferation, immunophenotypes and levels of homologous cytokine-induced killer cells (CIK) and the toxicity on leukemia cells.

Method: Mononuclear cell-induced DC-CIK cells derived from umbilical cord blood were collected and co-cultured in the proportion of 1:5. Cord blood CIK cells or peripheral blood DC-CIK cells were used as control. Phenotypes were analyzed by flow cytometry; vial cell counting was performed using trypan blue, and the killing activity of effector cells against leukemia cells was measured by MTT assay. The levels of interferon-r (IFN-r), tumor necrosis factor-a (TNF-α) and interleukin-12 (IL-12) were determined by ELISA.

Results: The proliferative capacity of DC-CIK cells was obviously improved compared with cord blood CIK cells and peripheral blood DC-CIK cells (P<0.05, P<0.05). During the co-culture of cord blood DC-CIK cells, the ratios of CD 3 (+) CD 8 (+) and CD 3 (+) CD 56 (+) cells were obviously higher than that of CIK cells under the same conditions (P<0.05). On day 3 of co-culture, the levels of IL-12, IFN-r and TNF-a in cultured supernatant of cord blood DC-CIK cells were all higher than those secreted by CIK cells cultured alone (P<0.01, P<0.05, P<0.05). When the effector to target ratio was 2.5-20:1, the killing effect of cord blood DC-CIK cells against each subtype of acute leukemia cells was obviously higher than that of CIK cells (P<0.05). No significant differences in killing effect were observed for different subtypes. This finding was consistent with the killing effect of peripheral blood DC-CIK cells against leukemia cells.

Conclusion: Cord blood DCs can enhance the proliferative capacity of homologous CIK cells and its anti-leukemia effect. Though cord blood DC-CIK cells showed a higher proliferative capacity than peripheral blood DC-CIK cells, the two types of DC-CIK cells did not differ significantly in terms of cytoxicity. With a high availability and the low probability of graft rejection reaction, cord blood DC-CIK cells have a brighter prospect for application in immunotherapy.

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