摘要
目的:探讨经典激活的巨噬细胞(Mφ1)和选择性激活的巨噬细胞(Mφ2)培养上清液对人γδT细胞增殖、表面标志、杀瘤活性的影响及其作用机制.方法:在体外用GM-CSF和IFN-γ诱导培养Mφ1,用M-CSF培养Mφ2,已戊烯焦磷酸法扩增外周血γδT细胞.用流式细胞术(FCM)检测巨噬细胞、γδT细胞表面标志,用酶联免疫吸附试验(ELISA)检测IL-10、IL-12含量,用四甲基偶氮唑蓝(MTT)法检测巨噬细胞培养上清对γδT细胞增殖的影响.用乳酸脱氢酶(LDH)释放法检测巨噬细胞培养上清对γδT细胞的杀瘤活性的影响.结果:体外诱导培养10d的Mφ1和Mφ2均高表达CD68(73.2%vs61.8%),Mφ1分泌IL-12的浓度显著高于Mφ2(35mg/Lvs9mg/L,P<0.01)Mφ1分泌IL-10的浓度明显低于Mφ2(15mg/Lvs87mg/L,P<0.01).不同浓度Mφ1上清组对γδT细胞的增值率均高于对照组或Mφ2组(338%vs11%,0,P<0.01).Mφ1培养上清作用后的γδT细胞表面标志γδTCR表达率高于Mφ2组和对照组,有统计学意义(97.3%vs89.1%91.3%,P<0.05).不同浓度Mφ1上清组对γδT细胞的杀瘤活性均高于对照组或Mφ2组(70.18%vs51.38%,47.25%,P<0.01).结论:Mφ1能够促进γδT细胞生长,而且能够提高γδT细胞对胃癌SGC-7901细胞的杀伤活性,Mφ2对γδT细胞作用不明显.
AIM：To investigate the effects of the culture supernatants of classically activated macrophages （Mφ1） and alternatively activated macrophages （Mφ2） on the proliferation, cytotoxicity, and surface maker expression of gamma delta T （γδT） cells and explore potential mechanisms involved. METHODS： Mφ1 were induced in vitro with granulocyte-macrophage colony-stimulating factor （GM-CSF） and interferon-γ （IFN-γ）, while Mφ2 were induced with macrophage colonystimulating factor （M-CSF）. The isopentenyl pyrophosphate method was used to amplify human peripheral blood γδT cells. The surface markers on macrophages and γδT cells were determined by flow cytometry （FCM）. Interleukin-10 （IL-10） and IL-12 levels in the culture supernatants of Mφ1 and Mφ2 were determined by enzymelinked immunosorbant assay （ELISA） using commercial kits. The proliferation of γδT cells induced with the culture supernatants of Mφ1 and Mφ2 was investigated by methyl thiazoly tetrazolium （MTT） assay. The lactate dehydrogenase （LDH） method was used to detect the cytotoxicity of γδT cells against gastric cancer SGC-7901 cells. RESULTS： After 10 days of induction culture, approximately 73.2% and 61.8% of Mφ1 and Mφ2 highly expressed CD68, respectively. The level of IL-12 secreted by Mφ1 was significantly higher than that secreted by Mφ2 （35 mg/L vs 9 mg/L, P〈 0.001）. The level of IL-10 secreted by Mφ1 was significantly lower than that secreted by Mφ2 （15 mg/L vs 87 mg/L, P 〈0.001）. The culture supernatant of Mφ1 could increase the proliferation of γδT cell when compared with those of Mφ2 and control cells （338% vs 11% and 0%, respectively; both P〈 0.01）. The positive rate of surface maker γδT cell receptor （γδTCR） on γδT cells induced with the culture supernatant of Mφ1 was higher than those on γδT cells induced with the culture supernatants of Mφ2 and control cells （97.3% vs 89.1% and 91.3%, respectively; both P 〈0.05）. The culture supernatant of Mφ1 could increase the cytotoxicity of γδT cells when compared with those of Mφ2 and control cells （70.18% vs 51.38% and 47.25%, respectively; both P〈 0.01）. CONCLUSION： The culture supernatant of Mφ1 can increase the proliferation and cytotoxicity of γδT cells, whereas the culture supernatant of Mφ2 has no significant effects.
出处
《世界华人消化杂志》
CAS
北大核心
2010年第1期20-27,共8页
World Chinese Journal of Digestology
