抗原负载DCs诱导的CIK细胞对B16黑色素瘤抑制作用的形态学观察

目的:探讨抗原负载树突状细胞(dentritic cells,DCs)诱导的CIK(cytokine induced killer)细胞对B16黑色素瘤的抑瘤作用。方法:分离、培养DC和CIK细胞,取部分DC进行肿瘤抗原负载,将其与CIK细胞按1:10的比例共培养3d,即为抗原负载的DC-CIK。建立B16黑色素瘤小鼠模型,分别于瘤周围皮下注射经Brdu标记的CIK、DC-CIK、抗原负载DC-CIK。按注射细胞进行分组,测量注射前后各组小鼠的瘤体积,计算抑瘤率,比较其抑瘤作用。应用免疫组化方法和透射电镜观察抗原负载DC-CIK细胞在皮肤中的分布及杀伤肿瘤细胞的形态学表现。结果:抗原负载DC诱导的CIK(细胞组抑瘤率(86.57%)高于CIK细胞组(33.34%,P<0.05)和DC-CIK细胞组(61.08%,P<0.05);光镜下抗原负载DC-CIK细胞主要分布在皮下组织,癌组织周围,特别是癌巢周边。透射电镜下抗原负载DC-CIK细胞体积大,核有切迹,细胞质内细胞器丰富,粗面内质网扩张。细胞表面有突起,与肿瘤细胞密切接触。大量肿瘤细胞凋亡、坏死。结论:CIK细胞经抗原负载DC诱导后抑瘤作用明显强于单纯CIK细胞和DC-CIK细胞。

Morphological Observation of Inhibition Effect on B16 Melanoma of CIK Cells Induced by DCs loaded with Tumor Antigen

Objective:To investigate the inhibitory effect of cytokine-induced ldller (CIK) cells on B16 melanoma cells induced by dendritic cells (DCs) loaded with tumor antigen.Methods:DCs and CIK cells were separated and cultured respective- ly,then DCs were loaded with tumor antigen;subsequently half of CIK cells were cultured and induced by DCs loaded with tumor antigen,while others continued to be co-culture with DCs.According to the change of tumor volume and the tumor inhibition rates,inhibitory effect of CIK cells on B16 melanoma cells was estimated.Distribution and morphological characteristics of immune cells in tumor tissue were observed by optical microscopy,electron microscopy and immunohistochemistry.Results:Tumor inhibi- tion rate of DC-CIK cells loaded with tumor antigen (86.57%) was higher than that of either CIK cells (33.34%,P<0.05) or DC- CIK cells (61.08%,P<0.05).DC-CIK cells with tumor antigen were mainly gathered in the subcutaneous tissue and the tumor tis- sue,especially around the cancer nest.The DC-CIK cells were bigger than the normal lymphocytes and notch always appears on the cellular nucleus observed under TEM.Plenty of active-functioned organelles such as dilated endoplasmic reticulum were ob- served in the cytoplasm of the DC-CIK.Protrusions on the surface of the DC-GIK cells contacted dosdy with the tumor cells. Apoptosis and necrosis of the cells could be widely observed in tumor tissues.Conclusion:Inhibitory effect of DC-CIK loaded with tumor antigen on B16 melanoma is stronger than that of CIK or DC-CIK.