mB7-1修饰的B16细胞体外诱导免疫效应

目的 探讨mB7-1基因修饰的B16细胞的体外抗瘤效应.方法 提取小鼠脾细胞mRNA,PCR法获得mB7-1片段,与pcDNA3连接,并进行酶切鉴定及测序.重组质粒经脂质体介导转染鼠B16细胞,检测基因表达,MTT法检测CTL杀伤作用.结果 完成mB7-1-pcDNA3重组载体连接及鉴定,基因测序与GenBank一致,并转染至B16细胞,检测到基因表达.经mB7-1修饰B16细胞可诱导淋巴细胞产生明显抑瘤效应.结论 应用本文方法 可获得mB7-1修饰B16细胞系,为研究其抗瘸效应莫定了基础.

mB7-1转染大鼠卵巢癌细胞诱导细胞免疫应答及体内致瘤性研究

为探讨转染mB7-1基因大鼠卵巢癌细胞系后体外诱导细胞免疫应答及其致瘤性,我们以逆转录病毒为载体将mB7-1基因转染入大鼠低分化卵巢上皮癌细胞株NuTu-19中,流式细胞仪检测mB7-1的表达.用同源淋巴细胞肿瘤细胞混合培养实验(MTLCs)测定淋巴细胞增殖指数,MTT法检测细胞毒淋巴细胞的增殖及其杀瘤活性.将NuTu-19/Neu、NuTu-19/mB7-1细胞分别接种于同源大鼠Fischer344皮下,观察mB7-1修饰的肿瘤细胞在动物体内的致瘤性.结果显示,mB7-1基因成功地转染入大鼠低分化卵巢上皮癌细胞株NuTu-19,流式细胞仪测定mB7-1基因呈阳性表达.NuTu-19/mB7-1体外刺激脾淋巴细胞增殖能力明显高于对照组细胞(P＜0.05).NuTu-19/mB7-1体外诱导的CTL较对照组对NuTu-19细胞的杀伤率显著增高(P＜0.01).两种细胞体外增殖能力无明显改变,NuTu-19/mB7-1组在Fischer344大鼠体内成瘤时间明显晚于对照组(P＜0.05).由此可见,mB7-1基因修饰的鼠卵巢癌细胞可体外诱导细胞免疫应答,接种动物后,实验瘤苗的致瘤性下降.

Objective: To investigate the cellular immunity response in vitro and the tumorigenecities in vivo of mB7-1 gene transfected murine ovarian cancer cell line. Methods: mB7-1 gene was transfected into the NuTu-19 cell line by retrovirus vector, and the expression of mB7-1 gene was confirmed by flow cytometry(FCM).NuTu-19/neo and NuTu-19/mB7-1 cells were injected subcutaneously into syngeneic Fischer 344 rats respectively, and their tumorigenecities were recorded. Proliferation indices of lymphocyte were assayed after syngenieic mixed tumor-lymphocyte cultures(MTLCs). The lysis activity of CTL toward tumor cells was determined using methyl thiazolyl tetrazolium(MTT) assay. Results: Successful transfection of mB7-1 gene into NuTu-19 cell line was comfirmed with FCM. In vitro study showed that there was no obvious changes in cell growth of gene transfected cell line, compared with the cell line NuTu-19. NuTu-19/mB7-1 cells could induce more effective proliferation of effector lymphocytes( P ＜ 0.05). The lysis activity of CTL activated by NuTu-19/mB7-1 was stronger than that of NuTu-19/neo ( P ＜ 0.01). Tumor sizes were smaller in the NuTu-19/mB7-1 receptance syngeneic Fischer 344 rats compared with those in the control group. Conclusion: mB7-1 genetically modified ovarian cancer cells could induce the cellular immunity response in vitro and the tumorigenecitiy of NuTu-19 cells was decreased after inoculation with the experimental vaccine.