OAZI-1蛋白复合物在小鼠体内诱导特异性抗肿瘤效应的研究
发布时间：2018-11-04 10:01
【摘要】：目的:在实验动物的水平上分析来源于肿瘤细胞的鸟氨酸脱羧酶抗酶抑制因子-1(Ornithine decarboxylase antizyme inhibitor-1,OAZI-1)蛋白复合物能否在小鼠体内诱导特异性抗肿瘤效应。方法:用包被有OAZI-1抗体的免疫磁珠从B16-F1小鼠黑色素瘤细胞中分离OAZI-1蛋白复合物,用此复合物免疫小鼠后,再在小鼠皮下接种B16-F1活细胞,然后观察接种瘤在小鼠体内的成瘤及生长状况。ELISA法用于检测免疫小鼠血清中IFN-γ含量。乳酸脱氢酶释放实验(LDH)检测免疫小鼠脾脏淋巴细胞对B16-F1细胞的杀伤效应。上述动物实验用原核表达纯化的OAZI-1蛋白和PBS免疫的小鼠作为对照。结果:与对照小鼠相比,接种OAZI-1蛋白复合物的小鼠脾淋巴细胞(效应细胞)对B16-F1黑素瘤细胞(靶细胞)具有更强的杀伤能力。在三种不同的效∶靶比下(10∶1、50∶1、100∶1),该组小鼠脾淋巴细胞对靶细胞的杀伤活性分别为46.2%、59.5%和92.5%,显著性高于接种纯化OAZI-1蛋白组(36.1%、26.8%和45.9%)和接种PBS组(24.6%、24.0%和27.2%)小鼠脾淋巴细胞。此外,接种OAZI-1蛋白复合物的小鼠血清中抗肿瘤细胞因子IFN-γ含量(538.3 pg/ml)也显著性高于接种纯化OAZI-1蛋白组(256.2 pg/ml)和接种PBS组(131.0 pg/ml)小鼠。上述方法免疫的小鼠在皮下再接种B16-F1活细胞后,免疫OAZI-1蛋白复合物组小鼠成瘤率为40%,而PBS组和纯化OAZI-1蛋白组小鼠成瘤率为100%,且接种瘤在OAZI-1蛋白复合物免疫小鼠体内生长更为缓慢。结论:从B16-F1肿瘤细胞中分离的OAZI-1蛋白复合物中可能含有肿瘤抗原,用此复合物接种小鼠能在实验动物体内诱导抗肿瘤免疫杀伤活性。
[Abstract]:Aim: to investigate whether ornithine decarboxylase inhibitor 1 (Ornithine decarboxylase antizyme inhibitor-1,OAZI-1) protein complexes derived from tumor cells can induce specific antitumor effects in mice at the experimental animal level. Methods: immunomagnetic beads coated with OAZI-1 antibody were used to isolate OAZI-1 protein complex from melanoma cells of B16-F1 mice. After immunizing mice with this complex, live B16-F1 cells were inoculated subcutaneously in mice. Then the tumorigenesis and growth of inoculated mice were observed. ELISA method was used to detect the content of IFN- 纬 in serum of immunized mice. Lactate dehydrogenase release assay (LDH) was used to detect the killing effect of spleen lymphocytes on B16-F1 cells in immunized mice. The above animal experiments used prokaryotic expression of purified OAZI-1 protein and PBS immunized mice as control. Results: compared with control mice, spleen lymphocytes (effector cells) inoculated with OAZI-1 protein complex had stronger cytotoxicity to B16-F1 melanoma cells (target cells). Under three different effects: 10: 1: 50: 110: 1, the murine splenic lymphocytes had cytotoxicity to the target cells by 59.5% and 92.5%, respectively. The spleen lymphocytes of mice inoculated with purified OAZI-1 protein (26.8% and 45.9%) and inoculated with PBS (24.60.24% and 27.2%) were significantly higher than those of inoculated with purified OAZI-1 protein group (26.8% and 45.9%). In addition, The content of antitumor cytokine IFN- 纬 (538.3 pg/ml) in serum of mice inoculated with OAZI-1 protein complex was significantly higher than that of mice inoculated with purified OAZI-1 protein group (256.2 pg/ml) and inoculated with PBS group (131.0 pg/ml). After the mice immunized with the above method were subcutaneously inoculated with B16-F1 living cells, the tumorigenic rate of mice immunized with OAZI-1 protein complex was 40%, while that of PBS group and purified OAZI-1 protein group was 100%. The growth of inoculated tumor was slower in mice immunized with OAZI-1 protein complex. Conclusion: the OAZI-1 protein complex isolated from B16-F1 tumor cells may contain tumor antigen, and inoculated with this complex can induce antitumor immunity in mice.
【作者单位】： 三峡大学医学院肿瘤微环境与免疫治疗湖北省重点实验室;
【基金】：国家自然科学基金项目(81372265)
【分类号】：R730.51
本文编号：2309487
[Abstract]:Aim: to investigate whether ornithine decarboxylase inhibitor 1 (Ornithine decarboxylase antizyme inhibitor-1,OAZI-1) protein complexes derived from tumor cells can induce specific antitumor effects in mice at the experimental animal level. Methods: immunomagnetic beads coated with OAZI-1 antibody were used to isolate OAZI-1 protein complex from melanoma cells of B16-F1 mice. After immunizing mice with this complex, live B16-F1 cells were inoculated subcutaneously in mice. Then the tumorigenesis and growth of inoculated mice were observed. ELISA method was used to detect the content of IFN- 纬 in serum of immunized mice. Lactate dehydrogenase release assay (LDH) was used to detect the killing effect of spleen lymphocytes on B16-F1 cells in immunized mice. The above animal experiments used prokaryotic expression of purified OAZI-1 protein and PBS immunized mice as control. Results: compared with control mice, spleen lymphocytes (effector cells) inoculated with OAZI-1 protein complex had stronger cytotoxicity to B16-F1 melanoma cells (target cells). Under three different effects: 10: 1: 50: 110: 1, the murine splenic lymphocytes had cytotoxicity to the target cells by 59.5% and 92.5%, respectively. The spleen lymphocytes of mice inoculated with purified OAZI-1 protein (26.8% and 45.9%) and inoculated with PBS (24.60.24% and 27.2%) were significantly higher than those of inoculated with purified OAZI-1 protein group (26.8% and 45.9%). In addition, The content of antitumor cytokine IFN- 纬 (538.3 pg/ml) in serum of mice inoculated with OAZI-1 protein complex was significantly higher than that of mice inoculated with purified OAZI-1 protein group (256.2 pg/ml) and inoculated with PBS group (131.0 pg/ml). After the mice immunized with the above method were subcutaneously inoculated with B16-F1 living cells, the tumorigenic rate of mice immunized with OAZI-1 protein complex was 40%, while that of PBS group and purified OAZI-1 protein group was 100%. The growth of inoculated tumor was slower in mice immunized with OAZI-1 protein complex. Conclusion: the OAZI-1 protein complex isolated from B16-F1 tumor cells may contain tumor antigen, and inoculated with this complex can induce antitumor immunity in mice.
【作者单位】： 三峡大学医学院肿瘤微环境与免疫治疗湖北省重点实验室;
【基金】：国家自然科学基金项目(81372265)
【分类号】：R730.51
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