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经链球菌菌体制剂OK-432刺激的树突状细胞对自然杀伤细胞增殖及功能的影响
引用本文:于丽梅,陈剑群,陈复兴,刘军权,周忠海,陈玲. 经链球菌菌体制剂OK-432刺激的树突状细胞对自然杀伤细胞增殖及功能的影响[J]. 中国免疫学杂志, 2012, 28(6): 517-521
作者姓名:于丽梅  陈剑群  陈复兴  刘军权  周忠海  陈玲
作者单位:1. 徐州医学院附属医院消化科,徐州,221002
2. 中国人民解放军第九七医院实验科,徐州,221004
摘    要:
目的:探讨经链球菌菌体制剂OK-432刺激的树突状细胞(Dendritic cells,DC)对自体自然杀伤细胞(Naturalkiller cells,NK)体外扩增和功能的影响。方法:将DC分为未成熟DC组和OK-432刺激的DC组,48小时后MTT法检测DC增殖情况,FCM检测DC表型CD80、CD83、CD86的表达情况;后将未成熟DC组和OK-432刺激的DC组分别与NK细胞以1∶1、1∶5、1∶10、1∶20、1∶40比例混合继续培养。0、2、4、6天时计算NK细胞扩增倍数;FCM检测NK细胞PFP、GraB、CD107a的表达;LDH释放法检测NK对HepG2细胞的杀伤活性。结果:OK-432(5μg/ml)使DC增殖达到最佳(30%),且能显著增强DC表型表达(P<0.05)。1∶5(OK-DC)组NK细胞扩增倍数达最大(P<0.05);1∶5(OK-DC)组能显著促进NK释放穿孔素(Pore-forming protein,PFP)、颗粒酶(Granzyme B,GraB)、CD107a(P<0.05);1∶5(OK-DC)组对HepG2细胞的杀伤活性(67.12±5.36)%达到最大(P<0.05)。结论:OK-432(终浓度5μg/ml)能促进DC增殖,并促进DC成熟;OK-DC与NK细胞共培养能以剂量依赖的方式增强NK细胞杀伤HepG2细胞功能,可能与增加NK扩增倍数和PFP、GraB、CD107a的表达有关。

关 键 词:OK-432  树突状细胞  自然杀伤细胞  过继免疫治疗

Expansion and function effects on natural killer cells by dendritic cells stimulated with OK-432(Picibanil)
YU Li-Mei , CHEN Jian-Qun , CHEN Fu-Xing , LIU Jun-Quan , ZHOU Zhong-Hai , CHEN Ling. Expansion and function effects on natural killer cells by dendritic cells stimulated with OK-432(Picibanil)[J]. Chinese Journal of Immunology, 2012, 28(6): 517-521
Authors:YU Li-Mei    CHEN Jian-Qun    CHEN Fu-Xing    LIU Jun-Quan    ZHOU Zhong-Hai    CHEN Ling
Affiliation:.Department of Gastroenterology,the Affiliated Hospital of Xuzhou Medical College,Xuzhou 221002,China
Abstract:
Objective:To investigate expansion and function effects on autologous natural killer(NK) cells by dendritic cells stimulated with OK-432(Picibanil).Methods:There were two groups for DC:immature DC group and OK-432 stimulated DC group,the proliferation of DC was detected by MTT method and the expression of DC phenotype(CD80,CD83,CD86)was detected by FCM after 48 hours;then NK cells cocultured with DC(immature DC group and OK-432 stimulated DC group) at ratios of 1∶1,1∶5,1∶10,1∶20 and 1∶40,the proliferation of NK cells was assayed at days 0,2,4 and 6;the expression of PFP,GraB and CD107a of NK was detected by FCM;the cell-killing activity of NK cells to HepG2 cells was detected by LDH release method.Results:OK-432(5 μg/ml) achieved optimal proliferation(30%) and can significantly enhance the expression of DC phenotype(P<0.05).The amplification multiples of 1∶5(OK-DC) group achieved the maximum(P<0.05);1∶5(OK-DC) group can significantly promote NK release PFP,GraB,CD107a(P<0.05);the killing activity of 1∶5(OK-DC) group to HepG2 cells(67.12 ± 5.36)% achieved the maximum(P<0.05).Conclusion:OK-432(final concentration of 5 μg/ml) can significantly promote DC proliferation and maturation;the co-culture of OK-DC and NK cells can enhance the killing effect of NK cells to HepG2 cells in a dose-dependent manner,and the mechanism may be associated with the elevated expansion of NK cells and the secretion of PFP,GraB,CD107a.
Keywords:OK-432  Dendritic cell  Natural killer cell  Adoptive immunotherapy
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