9L-rIL12-TK大鼠胶质瘤基因免疫疫苗的安全可控性体内实验研究

In vivo controllability study of 9L-rIL12-TK live glioma cell vaccine in mice

目的评估9L-rIL12-TK脑胶质瘤基因免疫活细胞疫苗在体内的安全可控性。方法将40只裸鼠随机分成3组,分别于右侧腋部皮下接种9L-rIL12-TK细胞(n=14)、9L-TK细胞(n=14)和9L细胞(n=12),7d后将各组裸鼠分成两亚组,实验组(n=8,8,6)给予更昔洛韦(GCV)干预实验,对照组(均n=6)给予生理盐水注射,连续7d;观察裸鼠瘤体生长情况和生存时间,60d后取瘤结节及全身脏器行组织学检查(苏木精-伊红染色)。结果①给予GCV干预后7d,9L-rIL12-TK实验组及9L-TK实验组裸鼠瘤体平均体积较相应对照组及9L组显著性缩小(均P<0.05)。60d时,8只9L-rIL12-TK实验组裸鼠瘤体均完全消退,且均无复发;8只9L-TK实验组裸鼠中,瘤体完全消退3只,瘤体缩小后再次增大5只;余亚组瘤体均呈增大趋势。②GCV干预后,9L-rIL12-TK实验组和9L-TK实验组与相应对照组比较,生存时间明显延长(P<0.05)。③9L-rIL12-TK实验组和9L-TK实验组无瘤裸鼠接种部位无异型性肿瘤细胞,余裸鼠瘤体内均见异型性肿瘤细胞;所有裸鼠相关脏器病理学检查均未见肿瘤远处转移征象。结论9L-rIL12-TK大鼠胶质瘤细胞基因免疫疫苗在裸鼠体内可得到安全有效的控制,使制备免疫原性强且安全有效的胶质瘤基因免疫活细胞疫苗策略成为可能;其基因免疫治疗效应有待进一步研究。

Objective Toevaluatethesafecontrollability of 9L-rIL12-TKlivegliomacellvaccineinmiceinvivo. Methods Forty BALB/C nude mice were randomly divided into 9L-IL 12-TK group （n = 14）, 9L-TK group （n = 14） and 9L group （n = 12）, and 9L-rlL 12-TK cells, 9L-TK cells and 9L cells were inoculated subcutaneously into the right axilla of the mice respectively. Seven days later, each group was randomly subdivided into experimental group （n = 8, 8, 6） with ganciclovir （GCV） administration, and control group （n = 6, 6, 6） with normal saline injection for 7 days. Tumor growth was measured, and survival time recorded. Sixty days later, the tissues were drawn from all tumor nodules and organs for histological examination. Results （1）The mean tumor volume more significantly shrank in 9L-IL12-TK＋GCV group and 9L-TK＋GCV group than in corresponding control group and 9L group （P 〈0.05） 7 days after the GCV administration. By 60 days, complete tumor regression was seen in all the 8 9L-rIL12-TK＋GCV mice, with no relapse, and in 3, but increased again after tumor shrinkage in 5 of 9L-TK＋GCV group, while in the rest subgroups, the tumor volume showed increase. （2）Compared with the control groups, survival times of the mice in 9L-IL12-TK＋GCV group and 9L-TK＋GCV group were significantly prolonged （P 〈0.05）. （3）No heterotypic cell was observed in the injection site in mice without tumors, while it was observed in all other mice with tumor. No distant metastasis was observed in pathological examination of relative organs. Condusion 9L-rIL 12-TK live glioma gene vaccine could be safely controlled in mice in vivo, which makes it possible to prepare live glioma gene vaccine with strong immunogenicity, safety and effectiveness, but its therapeutic effect remains to be studied further.