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目的 探讨人IL 4基因修饰对人脑胶质瘤的抑瘤效应及可能机制。方法 以逆转录病毒载体将人IL 4基因导入人脑胶质瘤细胞系SHG4 4细胞 ,用3H掺入法及流式细胞仪分析IL 4基因转染对人脑胶质瘤细胞增殖及细胞周期的影响 ;3H掺入法、51Cr释放法检测IL 4基因修饰瘤苗对健康人外周血单个核细胞 (PBMC)增殖、细胞毒性T细胞 (CTL)反应的影响。结果 与野生型瘤细胞比较 ,IL 4基因修饰瘤细胞增殖明显被抑制 (P <0 0 5 ) ,79 2 9%的瘤细胞滞留于G0 /G1期细胞 ,IL 4基因修饰瘤苗所诱导的PBMC增值反应及CTL杀伤活性分别是野生型瘤细胞的 3倍及 7倍 (P <0 0 5 ) ,并能促进瘤特异性CTL及其前体细胞分泌IL 2。用抗IL 4单克隆抗体 (Mab)可明显阻断其诱导的CTL反应 ,抗IL 2Mab则完全阻断此诱导反应。加入IL 4基因修饰瘤苗培养上清至野生型肿瘤培养中也能抑制肿瘤增殖、诱导PBMC增殖和CTL反应。结论 IL 4基因修饰可通过直接抑制脑胶质瘤细胞增殖 ,诱导PBMC增殖及CTL杀伤活性而发挥抗瘤效应 ,其对CTL杀伤活性的促进作用可能与诱导CTL及其前体细胞分泌IL 2有关
Objective To investigate the anti-tumor effect and possible mechanism of human IL 4 gene modification on human glioma. Methods Human IL 4 gene was introduced into human glioma cell line SHG4 4 by retroviral vector. The proliferation and cell cycle of human glioma cells were analyzed by 3H incorporation and flow cytometry 3H-incorporation method and 51Cr release method were used to detect the effects of IL 4 gene-modified vaccine on the proliferation and cytotoxic T lymphocyte (CTL) response of peripheral blood mononuclear cells (PBMCs) from healthy donors. Results Compared with wild type tumor cells, proliferation of IL 4 gene modified tumor cells was significantly inhibited (P <0.05), 79.29% of tumor cells were retained in G0 / G1 phase cells, IL 4 gene modified vaccine induced PBMC proliferation response and CTL killing activity were 3-fold and 7-fold higher than that of wild-type tumor cells (P <0 05), respectively, and promoted the secretion of IL 2 by tumor-specific CTL and its precursor cells. The anti-IL-4 monoclonal antibody (Mab) could obviously block the induced CTL reaction, while anti-IL2Mab completely blocked this induction reaction. Addition of IL 4 gene modified tumor vaccine supernatant to wild-type tumor culture also inhibited tumor proliferation, induced PBMC proliferation and CTL response. Conclusion The IL 4 gene modification can exert anti-tumor effect by directly inhibiting the proliferation of glioma cells, inducing PBMC proliferation and CTL killing activity. The promoting effect of IL 4 gene on the cytotoxic activity of CTL may be related to the induction of IL 2 secretion by CTL and its precursor cells