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目的:观察胸腺肽α1(Tα1)对结肠癌细胞裂解物(TuLy)负载DC(LyDC)的表型和功能的影响,以及二者联合应用对裸鼠结肠癌的治疗作用。方法:从健康人外周血单个核细胞中常规诱导培养未成熟DC(imDC),并负载TuLy后制备LyDC疫苗。Tα1体外刺激imDC、LyDC,流式细胞术(FCM)检测DC表型变化;ELISA法检测LyDC与自体T细胞共培养时,Tα1对LyDC分泌IL-12以及活化T细胞分泌IFN-γ水平的影响;MTT法检测LyDC经Tα1刺激后所诱导的细胞毒活性的变化。对HT-29结肠癌裸鼠模型行人源化T细胞免疫重建,观察LyDC与Tα1联合应用时的体内抗肿瘤效果。结果:Tα1刺激后的imDC、LyDC表型HLA-DR、CD80、CD86、CD83均上调(P<0.01);Tα1刺激组上清液中细胞因子IL-12和IFN-γ的水平较未刺激组增加(P<0.05,P<0.01);LyDC经Tα1刺激后诱导的CTL细胞毒活性较未经Tα1刺激组增强(P<0.01)。结肠癌裸鼠模型体内的人源化细胞免疫重建成功,在接种HT-29细胞58d后,LyDC+Tα1组和LyDC组的抑瘤率分别为60.41%和37.20%,二组之间瘤体积及瘤质量比较具有统计学意义(P<0.01)。结论:Tα1可促进DC分化成熟,并能增强LyDC诱导的CD4+Th1细胞反应和CTL的杀伤效应。Tα1与LyDC疫苗联合应用时具有较好的免疫佐剂活性和抗肿瘤作用。
Objective: To observe the effect of thymosin α1 (Tα1) on the phenotype and function of colon cancer cell lysate (TuLy) -loaded DC (LyDC) and the therapeutic effect of the combination of the two on colon cancer in nude mice. Methods: ImDC (imDC) was routinely induced from healthy human peripheral blood mononuclear cells and LyDC vaccine was prepared after loading TuLy. The effects of Tα1 on the secretion of IL-12 by LyDC and the secretion of IFN-γ by activated T cells were detected by ELISA assay when imDC, LyDC and FCM were used to detect the changes of DC phenotype. The cytotoxic activity of LyDC induced by Tα1 was detected by MTT assay. To investigate the anti-tumor effect of LyDC in combination with Tα1 in human colon cancer xenografts in nude mice. Results: The levels of HLA-DR, CD80, CD86 and CD83 in imDC and T lymphocytes were significantly increased after Tα1 stimulation (P <0.01). The levels of IL-12 and IFN-γ in supernatant of Tα1-stimulated group were significantly higher than those of untreated (P <0.05, P <0.01). The cytotoxicity of CTL induced by Tα1 stimulated by LyDC was higher than that without Tα1 stimulation (P <0.01). The successful immunization of human colon carcinoma xenografts in nude mice models showed that the tumor inhibition rates of LyDC + Tα1 group and LyDC group were 60.41% and 37.20% respectively after being inoculated with HT-29 cells for 58 days. The tumor volume, The tumor mass was statistically significant (P <0.01). Conclusion: Tα1 can promote the differentiation and maturation of DCs, and enhance the cytotoxicity of Lymph-induced CD4 + Th1 cells and CTL. Tα1 and LyDC vaccine combination has good immune adjuvant activity and anti-tumor effect.