抗原负载的树突细胞对细胞因子诱导的杀伤细胞的影响

The Effect of Dendritic Cells Pulsed with Antigens on Cytokines Induced Killer Cells

目的:研究树突细胞(DC)负载不同形式的抗原对其成熟和功能的影响,及DC对细胞因子诱导的杀伤细胞(CIK)增殖和功能的影响。方法:外周血单个核细胞(PBMC)经不同细胞因子作用后培养成DC和CIK细胞,将Bel7402细胞制备成肿瘤细胞冻融物、坏死肿瘤细胞及正常生长的肿瘤细胞,并作为不同状态的肿瘤抗原刺激DC,以流式细胞仪检测DC表面分子的表达。应用MTT法检测肿瘤抗原负载的DC刺激的CIK对肿瘤细胞的杀伤力。结果:负载肿瘤细胞冻融物的DC其表面CD1a、CD80、CD83、HLA-DR高表达,促进CIK增殖和杀伤力的功能最强;与正常肿瘤细胞共培养的DC细胞成熟程度受到抑制;负载Bel7402抗原的DC刺激的CIK对Bel7402有较强杀伤力,而对HerpG2杀伤力弱。结论:肿瘤细胞冻融物预刺激的DC成熟程度最高,诱导抗原特异性CIK细胞的功能最强,为临床DC瘤苗和CIK的应用提供了实验依据。

Objective: To research the effect of burdening different antigens on the maturation and function of dendritic cells, and the effect of DCs on cytokines induced killers' (CIKs) proliferations and functions. Methods: Human peripheral blood monocyte (PBMC) were cultured into DCs and CIKs with different cytokines. The tumor cell line Bel 7402 prepared into carcinoma cell lysates、necrotic tumor cells and normally grew tumor cells were burdened on DCs as different tumor antigens and the DCs were used to stimulate CIKs. Through the analysis of DCs' surface molecules the detection of the proliferation and anti-tumor function of CIKs activated by DCs, we researched the influence of different antigens on DCs' maturation and function. Results: DCs pulsed with carcinoma cell lysates had the strongest power of promoting the proliferation and anti-tumor function of CIKs and had the highest expressed surface molecules as CD1a、CD80、 CD83、 HLA-DR; while the DCs co-cultured with normally grew tumor cells had the lowest maturation degree and function of stimulating CIKs; CIKs stimulated by DCs burdening on Bel 7402 antigens had relatively strong anti-tumor power on Bel 7402 while there was weak power on HerpG2. Conclusion: DCs pulsed with carcinoma cell lysates had the highest maturation degree and the strongest power of inducing CIKs, and it provided an experimental basis for the clinical use of CIKs.