分枝杆菌Ag85B/IL-2融合蛋白激活的人PBMC对膀胱肿瘤细胞毒效应的研究

Research on the Cytotoxicity of Human Peripheral Blood Mononuclear Cells Activated by Ag85B/IL-2 Fusion Protein Against Bladder Tumor Cells

【目的】研究分枝杆菌Ag85B/IL-2融合蛋白激活的人外周血单个核细胞(PBMC)对膀胱肿瘤的细胞毒效应。【方法】MTT法测定Ag85B/IL-2融合蛋白激活的效应细胞(AIAK)对四种膀胱肿瘤细胞BIU-87、T24、EJ、BTT739的细胞毒活性,并与单纯用Ag85B蛋白、BCG和rIL-2激活的效应细胞(分别命名为AAK、BAK和LAK)作比较;用流式细胞仪测定AIAK作用于四种膀胱肿瘤细胞后的凋亡水平。【结果】效靶比为10∶1时,AIAK对BIU-87的细胞毒作用显著高于LAK(P<0.05)和BAK(P<0.05);对T24的细胞毒作用非常显著高于AAK(P<0.01)和BAK组(P<0.01);对EJ细胞和BTT739细胞的细胞毒作用显著高于LAK(P<0.01)、AAK(P<0.01)和BAK组(P<0.01)。效靶比为20∶1时,AIAK对BIU-87、T24、E-J和BTT739四种膀胱肿瘤细胞的细胞毒作用均显著高于LAK、AAK和BAK组(P<0.05,P<0.01)。AIAK作用于四种膀胱肿瘤细胞后,BIU-87(P<0.05)、T24(P<0.05)、E-J(P<0.05)和BTT739(P<0.05)的凋亡水平均有显著上升。【结论】Ag85B/IL-2融合蛋白激活的人PBMC对膀胱肿瘤具有较高的细胞毒活性并促进其凋亡。

[Objective]To research the cytotoxicity of human peripheral blood mononuclear cells（PBMC） activated by Ag85B/IL-2 fusion protein against bladder tumor cells. [Methods]By assay of MTT, the cytotoxicity of human PBMC activated by Ag85B/IL-2 fusion protein against bladder tumor cells BIU-87,T24,EJ and BTT739 was studied , and compared with that of AAK,BAK and LAK which were activated by Ag85B protein,BCG and rIL-2, respectively. In addition, the apoptosis level of the bladder tumor cells acted by AIAK was observed with flow cytometer. [Results]When the rate of effector cells： target cells was 10 ： 1 , the cytotoxicity of effector cell of AIAK on BIU-87 was significantly higher than that of BAK （ P〈0.05） and LAK （ P 〈0.05）, and cytotoxicity on T24 was significantly higher than that of AAK （ P 〈0.01） and BAK （ P 0.01）, and cytotoxicity on EJ and BTT739 were significantly higher than that of LAK（ P 〈0.01）, AAK（ P 〈0.01） and BAK （ P 〈0.01）. When the rate of effector cells：target cells was 20 ： 1 , the cytotoxicity of ellector cell of AIAK on the four target cells was significantly higher than that of the four control groups（ P〈0.05 or P 〈0.01）. The apoptosis levels of BIU-87, T24 , E-J and BTT739 were significantly raised after acted by AIAK （all P 〈0.05, respectively）. [Conclusion]PBMC activated by Ag85B/IL-2 has cytotoxicity against bladder tumor cells and may promote its apoptosis level.