CIK细胞的生物学特性及荷瘤鼠体内分布特点研究

Biological characteristics and distributional regularity of CIK cells in the mice bearing tumor

目的:探讨细胞因子诱导的杀伤细胞(CIK细胞)在荷瘤鼠体内的分布规律。方法:用MTT比色法测定CIK细胞对Hela-luc细胞的杀伤作用。以人宫颈癌细胞Hela-luc接种BALB/c裸鼠建立动物模型。体外培养CIK细胞后以荧光染料CFSE标记,经不同途径注射入荷瘤鼠体内。用激光共聚焦显微镜和流式细胞技术检测方法分析CIK细胞在各器官的分布情况。结果:CIK细胞在体外培养14～18天时,细胞数量生长达高峰,此时CD3+CD56+T细胞的数量也达最高值。以10∶1、20∶1、40∶1的比例将效、靶细胞作用48小时后,CIK细胞对Hela-luc细胞的杀伤率分别为(24.08±3.18)%、(51.16±2.64)%、(72.14±4.21)%。CIK细胞经腹腔注射,肿瘤组织24小时达到最高(20.56%),瘤旁注射,肿瘤组织3小时达到最高(25.75%)。结论:CIK细胞在体外对宫颈癌Hela-luc细胞有较强的杀伤作用。CIK细胞在体内分布于肺、肝、肿瘤部位,其分布在各脏器中浓度规律与不同的输注途径有一定关系。针对不同解剖部位的肿瘤可以选择不同的CIK细胞输注途径以最大限度地提高疗效。

Objective：To investigate the distributional regularity of cytokine-induced killer cells （CIK cells） in mice bearing tumor.Methods：Killing activity of CIK cells to Hela-luc cells was measured by MTT assay.The BALB/c mouse model was successfully established by inoculating Hela-luc cells.The CIK cells isolated from peripheral blood mononuclear cells were labeled with CFSE,and then the labeled cells were injected into the mice bearing tumor via peritoneal cavity or pericancerous injection.The distribution of CIK cells in organs were detected by flow cytometry and laser scanning confocal microscope.Results：CIK cells grew up to the peak on the 14th day to 18th day,and the percentage of CD3＋CD56＋ T cells also reached to the maximum simultaneously.After the effect/target cells were treated for 48 h with the ratios of 10：1,20：1 and 40：1,the killing ratios of CIK cells to Hela-luc cells were （24.08±3.18）%,（51.16±2.64）% and （72.14±4.21）%,respectively.The concentration of CIK cells reached to the maximum （20.56%） in the tumor tissue 24 h after peritoneal cavity injection,and reached to the maximum （25.75%） 3 h after pericancerous injection.Conclusion：The CIK cells have a stronger killing effect on cervical cancer Hela-luc cells in vitro.The CIK cells are distributed in different tissues in vivo including lung,liver,spleen and tumor tissue,and the distributional regularity of the cells is related with the injection ways.Different injection ways could be selected for CIK cells based on the site of tumor in order to enhance the therapeutic effect to the maximum.