Raw264.7细胞蛋白和PD-L1抗体在小鼠黑色素瘤(B16)中的抑瘤效果评价

目的探讨小鼠来源白血病毒诱导的肿瘤细胞(Raw264.7)蛋白和程序性死亡-配体(PD-L1)在C57BL/6小鼠黑色素瘤(B16)中的抑瘤效果。方法采用6~8周龄清洁级C57BL/6小鼠30只(雌雄各半),平均分为3组。各组小鼠分别于皮下接种B16细胞(2×10⁵个/只),待小鼠肿瘤平均体积达到100 mm³时,给予治疗。group A:腹腔注射Raw264.7细胞蛋白(50μg/只),每周1次,共2次。group B:腹腔注射PD-L1抗体(600μg/只,本课题组生产纯化),每3天1次,共4次。group C:相同时间点给予等体积生理盐水对照。各组小鼠给药后,测量肿瘤大小,末次测量后,各组小鼠无菌取脾脏,应用流式细胞术分析CD3/4/8⁺T细胞群比例。结果各组荷瘤小鼠给予相应治疗后,组C小鼠肿瘤体积增长迅速,末次测量平均体积达到4004 mm³;组B小鼠增长次之,末次平均体积达到1637 mm³;组A小鼠体积增长最慢,末次平均体积仅为882 mm³(P<0.01)。对小鼠脾脏免疫细胞分析显示,组B小鼠中CD3⁺CD8⁺T cell比例明显上调,平均达到77.65%,而CD3⁺CD4⁺T比例明显下调,仅为11.86%(P<0.01);而组A和组C两组小鼠CD3⁺CD4⁺T和CD3⁺CD8⁺T没有明显区别(P>0.05)。结论制备的RAW264.7细胞灭活蛋白和PD-L1抗体对于B16荷瘤小鼠均有抑瘤作用,且前者效果更为明显。

Evaluation of Anti-tumor Effect of Raw264.7 Cell Protein and PD-L1 Antibody in Mouse Melanoma(B16)

Objective To investigate the tumor suppressive effect of mouse leukemia-induced tumor cell(Raw264.7)protein and programmed death-ligand(PD-L1)in C57BL/116 mouse melanoma(B16).Methods Thirty clean C57BL/116 mice(half male and half male)aged 6-8 weeks were divided into three groups on average.The mice in each group were inoculated with B16 cells(2×10⁵cells/mouse)subcutaneously,then were treated when the average tumor volume reached 100 mm³.Group A:Raw264.7 cell protein(50μg/piece)was injected intraperitoneally once a week for two times.Group B:Intraperitoneal injection of PD-L1 antibody(600μg/body,produced and purified by our group),once every three days,a total of four times.Group C:The same volume of saline control was given at the same time point.After administration of each group of mice,the tumor size was measured.After the last measurement,the mice of each group were aseptically taken for spleen and analyzed for the proportion of CD3/4/8⁺T cell by flow cytometry.Results After the corresponding treatment of tumor-bearing mice in each group,the tumor volume of group C mice grew rapidly,and the average volume of the last measurement reached 4004 mm³;the growth of group B mice was second,and the average volume of the last time reached 1637 mm³;the volume growth of group A mice was the most Slow,the last average volume is only 882 mm³(P<0.01).Analysis of mouse spleen immune cells showed that the proportion of CD3⁺CD8⁺T cells in Group B mice was significantly increased,reaching an average of 77.65%,while the proportion of CD3⁺CD4⁺T was significantly decreased,only 11.86%(P<0.01);There were no significant differences between group A and group C,CD3⁺CD4⁺T and CD3⁺CD8⁺T(P>0.05).Conclusion The RAW264.7 cell inactivation protein and PD-L1 antibody prepared by this research group have anti-tumor effect on B16 tumor-bearing mice,and the former effect is more obvious.