树突状细胞负载丁酸梭菌诱导免疫效应细胞对裸鼠皮下BxPC-3移植瘤的抑制作用

Dendritic cells loaded with Clostridium butyricum induce immune effector cells to inhibit subcutaneous BxPC-3 transplanted tumor in nude mice

目的探讨丁酸梭菌(CB)致敏树突状细胞(DCs)诱导免疫效应细胞(IECs)对BALB/c裸鼠皮下BxPC-3移植瘤的抑制效果。方法BALB/c裸鼠安全性评价实验:28只裸鼠随机分为7组,实验第1天,取对数生长的BxPC-3注射于裸鼠左下臂,14 d后测量各组裸鼠皮下移植瘤大小,空白对照G组注射无菌生理盐水,A~F组分别皮下注射不同浓度CB致敏的DCs和IECs。49 d后测量移植瘤的大小。BALB/c裸鼠体内效能实验:28只BALB/c裸鼠随机为7组,实验第1天,取对数生长的BxPC-3注射裸鼠左下臂。14 d后,G组裸鼠左下臂移植瘤注射无菌生理盐水,A~F组分别皮下注射不同浓度CB致敏的DCs和IECs。49 d后处死裸鼠并分离皮下移植瘤,通过苏木精-伊红(HE)染色法观察裸鼠皮下移植瘤病理切片,电子秤称其重量并计算抑瘤率。采用单因素方差分析。结果(1)BALB/c裸鼠生物安全性评价实验结果。A组(0.99±0.27)cm^(2)最大,F组(0.45±0.20)cm^(2)最小,其他组为B组>C组>D组>E组,其中D、E、F组与G组比较差异有统计学意义(F=5.134,P<0.05)。(2)BALB/c裸鼠生物效能实验结果。A组(1.24±0.26)g,F组(0.77±0.22)g,其他组为B组>C组>D组>E组,其中D、E、F组与G组比较差异有统计学意义(F=7.364,P<0.05)。同时,F组抑瘤率高达46.52%,A组抑瘤率最低13.8%,其他组皮下移植瘤抑制率为B组<C组<D组<E组。结论CB致敏的DCs具有良好的生物安全性和诱导IECs抗BxPC-3移植瘤作用。

Objective To discuss the inhibitory effect of immune effector cells(IECs)induced by dendritic cells sensitized by Clostridium butyricum(CB)on subcutaneous BxPC-3 transplanted tumor in BALB/c nude mice.Methods Safety evaluation experiment of BALB/c nude mice:a total of 28 nude mice were randomly divided into 7 groups.On the first day of the experiment,log-grown BxPC-3 was injected into the left lower arm of the nude mice.After 14 days,the subcutaneous xenografts size of nude mice in each group was measured.The blank control group G was injected with sterile normal saline,and the groups A-F were respectively injected with different concentrations of CB-sensitized DCs and IECs subcutaneously.The size of the xenografts was measured after 49 days.Efficacy experiment of BALB/c in nude mice:a total of 28 BALB/c nude mice were randomly divided into 7 groups.On the first day of the experiment,log-grown BxPC-3 was injected into the left lower arm of the nude mice.After 14 days,the xenografts in the left lower arm of the nude mice in group G were injected with sterile normal saline,while those in group A-F were injected with different concentrations of CB-sensitized DCs and IECs subcutaneously,respectively.After 49 days,the nude mice were sacrificed and the subcutaneous xenografts were isolated.The pathological sections of the subcutaneous xenografts in nude mice were observed by hematoxylin and eosin(HE)staining,and the weight was measured by electron microscopy to calculate the tumor inhibition rate.SPSS 21.0 statistical software was used for analysis,and single-factor analysis of variance was used.The data were expressed by mean±standard deviation(Mean±SD),and the difference was statistically significant with P<0.05.Results(1)Biosafety evaluation experiment of BALB/c nude mice:The tumor size in group A[(0.99±0.27)cm^(2)]was the largest,that in group F[(0.45±0.20)cm^(2)]was the smallest,and that in the rest groups was group B>group C>group D>group E.There were significant differences between groups D,E or F and G(F=5.134,P<0.05).(2)Bioefficiency experiment of BALB/c nude mice:The weight in group A was(1.24±0.26)g,that in group F was(0.77±0.22)g,and that in the rest groups was group B>group C>group D>group E.The differences between groups D,E or F and G were statistically significant(F=7.364,P<0.05).Conclusion CB-sensitized DCs had good biosafety and induced IECs against BxPC-3 xenografts.