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目的:研究共刺激分子4-1BBL分子逆向信号对原代急性髓细胞白血病细胞的促增殖作用及其生物学效应。方法:用流式细胞术(FCM)检测4-1BBL分子在26例初诊急性髓细胞白血病(AML)患者白血病细胞表面的表达,分离AML患者的白血病细胞并加入激发型4-1BBL抗体(1F1单抗)共培养,同时设置IgG1同型对照组,Alamar Blue法检测4-1BBL单抗1F1促AML原代细胞的增殖。收集培养细胞的上清液,用ELISA方法检测基质金属蛋白酶-9(MMP-9)的水平。结果:FCM测得初诊AML患者的白血病细胞上存在4-1BBL分子的表达,26例AML患者中4-1BBL分子的阳性表达率为46.15%;1F1单抗能明显促进4-1BBL+-AML细胞的生长,4-1BBL+-AML组刺激指数明显高于4-1BBL--AML组(P<0.01)。1F1与4-1BBL+-AML细胞共培养48 h后,用ELISA法检测其上清液中MMP-9含量显著高于IgG1同型对照组[(965.06±229.94)vs(596.49±129.28)pg/ml,P<0.01],而4-1BBL--AML组的MMP-9与IgG1组无显著差异(P>0.05)。结论:部分AML患者的白血病细胞可以表达4-1BBL分子,1F1单抗与AML细胞上4-1BBL分子交联后,可以促进AML细胞的生长,同时促进MMP-9的分泌。
OBJECTIVE: To study the pro-proliferative effect and biological effects of 4-1BBL molecular costimulatory molecules on primary acute myeloid leukemia cells. Methods: The expression of 4-1BBL on 26 leukemia cells in newly diagnosed acute myeloid leukemia (AML) was detected by flow cytometry (FCM). Leukemic cells from AML patients were isolated and stimulated with 4-1BBL antibody (1F1 single Anti-co-culture, while setting IgG1 isotype control group, Alamar Blue assay 4-1BBL monoclonal antibody 1F1 primary proliferation of AML cells. The supernatant of the cultured cells was collected and the level of matrix metalloproteinase-9 (MMP-9) was detected by ELISA. Results: The expression of 4-1BBL was detected on the leukemia cells of newly diagnosed AML patients by FCM. The positive rate of 4-1BBL in 26 AML patients was 46.15%. The 1F1 monoclonal antibody could significantly promote 4-1BBL + -AML cells The stimulation index of 4-1BBL + -AML group was significantly higher than that of 4-1BBL - AML group (P <0.01). After co-cultured with 4-1BBL + -AML cells for 48 h, the content of MMP-9 in the supernatant of 1F1 was significantly higher than that in the IgG1 isotype control group [(965.06 ± 229.94) vs (596.49 ± 129.28) pg / ml, P <0.01]. There was no significant difference between MMP - 9 and IgG1 in 4-1BBL - AML group (P> 0.05). Conclusion: The leukemia cells in some AML patients can express 4-1BBL molecules. After 1F1 monoclonal antibody cross-linked with 4-1BBL molecules on AML cells, it can promote the growth of AML cells and promote the secretion of MMP-9.