IL-23诱导人外周血单个核细胞增殖及其体外杀伤MUTZ-1细胞的实验研究  

Experimental study on the effects of IL-23 in inducing proliferation of human peripheral blood mononuclear cell and in killing MUTZ-1 cells in vitro

作　　者：王瑞仓[1] 刘艳芬[2] 杨洁[1] 周洁[1] 李杰[1] 郝洪岭[1] WANG Ruicang LIU Yanfen YANG Jie(Department of Hema＇tology, Hebei Provincial People＇ s Hospital, Shijiazhuang 050051, China)

机构地区：[1]河北省人民医院血液科,石家庄市050051 [2]河北省唐山市人民医院血液科

出　　处：《河北医药》2016年第20期3055-3058,3062,共5页Hebei Medical Journal

基　　金：河北省医学科学研究重点课题计划(编号:ZD20140444)

摘　　要：目的观察IL-23诱导人外周血单个核细胞(PBMNC)增殖及其对骨髓增生异常综合征(MDS)细胞系MUTZ-1的杀瘤作用。方法采用密度梯度离心法分离获得正常人PBMNC,细胞分4组:阴性对照组(未加IL-23),3个浓度IL-23组(2、10、50 ng/ml),体外诱导72 h,并与MUTZ-1共培养。流式细胞术检测不同时间诱导后PBMNC细胞增殖和表型变化。乳酸脱氢酶(LDH)释放法检测PBMNC对MUTZ-1细胞的杀瘤效果。Real time-PCR和Westernblot检测PBMNC细胞培养上清液中IFN-γ、TNF-α、TGF-β、颗粒酶A、颗粒酶B、穿孔素mRNA和蛋白表达。结果体外诱导培养1、3、5、7 d时,IL-23诱导能够明显促进PBMNC细胞增殖,呈时间和浓度依赖性,与阴性对照组比较差异有统计学意义(P<0.05);CD+3、CD+4、CD+56阳性细胞数明显增加,CD+8阳性细胞数明显降低,且IL-23作用后的PBMNC均对MUTZ-1细胞产生良好的杀瘤活性;与阴性对照组比较,IL-23作用后PBMNC细胞上清液中IFN-γ、TNF-α、TGF-β、颗粒酶A、颗粒酶B、穿孔素mRNA和蛋白表达明显增加,呈时间和浓度依赖性,差异有统计学意义(P<0.05)。结论 IL-23能够明显促进PBMNC细胞增殖,并通过诱导PBMNC细胞表达IFN-γ、TNF-α、TGF-β、颗粒酶A、颗粒酶B、穿孔素发挥杀伤MUTZ-1细胞作用。Objective To observe the effects of IL -23 in inducing proliferation of human peripheral blood mononuclear cell （ PBMNC） and in killing MUTZ-1 cells—a human myelodysplastic syndrome （ MDS） cell line in vitro. Methods The PBMNCs were separated by ficoll density gradient centrifugation.The experiment was divided into control group （without IL-23）, IL-23 high,median,low dose groups （2, 10, 50ngm/l,respectively） in which the PBMNCs were induced by IL-23 for 72 hours in vitro.Then PBMNCs were co-cultured with MUTZ-1 cells.The proliferative ability of PBMNC cells was detected by flow cytometry, and the killing effect of PBMNC on MUTZ-1 cells was measured by LDH releasing method . Moreover the expression levels of IFN-γ, TNF-α, TGF-β, granzyme A, granzyme B and perforin at mRNA and protein levels were detected by Real time-PCR and Western Blot,respectively.Result s After 1, 3, 5,7 -day induction, IL-23 could obviously promote the proliferation of PBMNCs,with a concentration and time-dependent manner, as compared with that in control group （ P 〈0.05）.The positive cell numbers of CD3＋, CD4＋and CD5＋6 were significantly increased, however, CD8＋cells were obviously decreased,moreover,the killing effects of PBMNC induced by IL-23 on MUTZ-1 cells were increased. After treated by IL-23, the expression levels of IFN-γ, TNF-α, TGF-β, granzyme A, granzyme B and perforin at mRNA and protein levels were significantly increased , with a concentration and time -dependent manner, as compared with those in control group （ P 〈0.05）.Conclusion IL -23 can obviously promote the proliferation of PBMNCs and can kill MUTZ-1 cells through inducing the expressions of IFN-γ, TNF-α, TGF-β, granzyme A, granzyme B and perforin of PBMNCs.

关 键 词：IL-23 MUTZ-1细胞 杀瘤活性 

分 类 号：R551.31[医药卫生—血液循环系统疾病]