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转基因马铃薯中人白介素-12的生物学活性 被引量:1

Bioactivity of human interleukin-12 from transgenic potatoes
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摘要 目的检测转基因马铃薯体系中表达的人白介素-12(human interleukin 12,hIL-12)的肿瘤杀伤活性。方法通过ELISA法检测转基因马铃薯中hIL-12刺激外周血单个核细胞(PBMC)产生IFN-γ的量以反映生物学活性,并观察hIL-12单克隆抗体对此生物活性的阻断效应;MTT法检测经转基因马铃薯中hIL-12作用的PBMC对NK敏感肿瘤细胞K562的杀伤作用,肿瘤细胞凋亡以Annexin V/PI双染后激光共聚焦显微镜观察。结果转基因马铃薯叶和块茎中表达的hIL-12与重组人白介素-12(rhIL-12)标准品在25~400 pg/ml间均能诱生IFN-γ,其中转基因马铃薯叶和块茎中hIL-12刺激PBMC产生IFN-γ的量分别为(163.34±22.17)~(12 107.10±3 996.51)pg/ml和(224.20±25.76)~(11 196.30±4 067.48)pg/ml,与野生对照组均有统计学差异(P<0.01),而hIL-12单克隆抗体能完全阻断此诱生IFN-γ效应。同时200 pg/ml转基因马铃薯叶、块茎中提取的hIL-12对人红白血病细胞K562的杀伤率分别为(28.4±8.7)%、(32.3±6.9)%,而与野生对照组(10.3±4.9)%的杀伤率相比,有统计学差异(P<0.05)。激光共聚焦显微镜观察到经转基因马铃薯表达hIL-12作用的K562细胞出现早、中、晚各期的凋亡特征。结论转基因马铃薯表达的hIL-12具有诱导PBMC产生IFN-γ及杀伤肿瘤细胞K562的生物学活性。 Objective To detect the bioactivity of human iuterleukin-12 (hlL-12) extracted from transgenic potatoes. Methods ELISA was used to assess hIL-12-induced IFN-γ secretion in peripheral blood mononuclear cells (PBMC) and observe the blocking effect of monoclonal antibody to hIL-12 on the bioactivity of hIL-12. Lethal effect of hIL-12-activated PBMC on NK sensitive I(562 cell line was detected by MTT assay. Apoptosis of tumor cells was observed under laser confocal microscope with double Annexin V/PI staining. Results The expression of hlL-12 and recombinant hIL-12 (rhlL-12) extracted from transgenic potato leaves or tubers at the concentration of 25 pg/ml to 400 pg/ml induced IFN-3, secretion in PBMC. The mount of secreted IFN-γ/was (163.34± 22.17 )pg/ml to (12 107.10 ±3 996.51 )pg/ml after treated with hIL-12 from potato leaves and was (224.20 ± 25.76 ) pg/ml to ( 11 196.30 ± 4 067.48 ) pg/ml after treated with potato tubers, which was significantly higher than that of commercially available hlL-12 (P 〈0.01 ). The secreted IFN-γ/was almost fully blocked by monoclonal hIL-12 antibody. The apoptotic rate of K562 cells treated with 200 pg/ml hIL-12 extracted from transgenic potato leaves or tubers was (28.4 ± 8.7)% and (32.3±6.9)%, respectively, which was much higher than that of commercially available hIL-12 (P 〈 0.05 ). Laser confocal microscopy showed the characteristic apoptosis of K562 cells at early, middle and late phases after treated with the extracted hIL-12. Conclusion hIL-12 extracted from transgenic potato induces PBMC producing IFN-γ and excert antitumor effect on K562 leukemia cells.
出处 《第三军医大学学报》 CAS CSCD 北大核心 2010年第15期1632-1634,共3页 Journal of Third Military Medical University
基金 贵州省科委重大攻关项目(2004NGZ002) 贵州省教育厅重点项目(2004118)~~
关键词 人白介素-12 生物活性 马铃薯 转基因植物 human interleukin 12 bioactivity potato transgenic plant
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共引文献14

同被引文献27

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