【摘要】：目的:在實驗動物的水平上分析來源于腫瘤細胞的鳥氨酸脫羧酶抗酶抑制因子-1(Ornithine decarboxylase antizyme inhibitor-1,OAZI-1)蛋白復合物能否在小鼠體內誘導特異性抗腫瘤效應。方法:用包被有OAZI-1抗體的免疫磁珠從B16-F1小鼠黑色素瘤細胞中分離OAZI-1蛋白復合物,用此復合物免疫小鼠后,再在小鼠皮下接種B16-F1活細胞,然后觀察接種瘤在小鼠體內的成瘤及生長狀況。ELISA法用于檢測免疫小鼠血清中IFN-γ含量。乳酸脫氫酶釋放實驗(LDH)檢測免疫小鼠脾臟淋巴細胞對B16-F1細胞的殺傷效應。上述動物實驗用原核表達純化的OAZI-1蛋白和PBS免疫的小鼠作為對照。結果:與對照小鼠相比,接種OAZI-1蛋白復合物的小鼠脾淋巴細胞(效應細胞)對B16-F1黑素瘤細胞(靶細胞)具有更強的殺傷能力。在三種不同的效∶靶比下(10∶1、50∶1、100∶1),該組小鼠脾淋巴細胞對靶細胞的殺傷活性分別為46.2%、59.5%和92.5%,顯著性高于接種純化OAZI-1蛋白組(36.1%、26.8%和45.9%)和接種PBS組(24.6%、24.0%和27.2%)小鼠脾淋巴細胞。此外,接種OAZI-1蛋白復合物的小鼠血清中抗腫瘤細胞因子IFN-γ含量(538.3 pg/ml)也顯著性高于接種純化OAZI-1蛋白組(256.2 pg/ml)和接種PBS組(131.0 pg/ml)小鼠。上述方法免疫的小鼠在皮下再接種B16-F1活細胞后,免疫OAZI-1蛋白復合物組小鼠成瘤率為40%,而PBS組和純化OAZI-1蛋白組小鼠成瘤率為100%,且接種瘤在OAZI-1蛋白復合物免疫小鼠體內生長更為緩慢。結論:從B16-F1腫瘤細胞中分離的OAZI-1蛋白復合物中可能含有腫瘤抗原,用此復合物接種小鼠能在實驗動物體內誘導抗腫瘤免疫殺傷活性。
[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- 緯 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- 緯 (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.
【作者單位】： 三峽大學醫(yī)學院腫瘤微環(huán)境與免疫治療湖北省重點實驗室;
【基金】：國家自然科學基金項目(81372265)
【分類號】：R730.51

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