现代肿瘤医学
現代腫瘤醫學
현대종류의학
JOURNAL OF MODERN ONCOLOGY
2015年
18期
2569-2572,2573
,共5页
树突状细胞%K562 细胞%白细胞介素 12%转化生长因子 β1%血管内皮生长因子
樹突狀細胞%K562 細胞%白細胞介素 12%轉化生長因子 β1%血管內皮生長因子
수돌상세포%K562 세포%백세포개소 12%전화생장인자 β1%혈관내피생장인자
dendritic cells%K562 cells%IL -12%TGFβ1%VEGF
目的：探讨共培养条件下血液肿瘤 K562细胞和树突状细胞（dendritic cells，DCs）在表达细胞因子方面的相互作用情况，探索血液肿瘤发生发展过程中的免疫逃逸机制。方法：利用免疫磁珠试剂盒从人健康外周血中分离纯化出高纯度的 CD14＋单核细胞，一定浓度的巨噬细胞集落刺激因子（granlocyte －macrophage co-lomy stimulating factor，GM－CSF）和白介素4（interlenkins －4，IL －4）将单核细胞诱导分化为未成熟 DCs（im-mature DCs，imDCs），再用一定浓度的肿瘤坏死因子（tumor necrosis factor －α，TNF －α）将 imDCs 诱导为成熟DCs（mature DCs，mDCs），分别将 imDCs 和 mDCs 与 K562细胞在 Transwell 系统中共培养48h，对照组为正常培养48h 的 DCs 和撤细胞因子培养48h 的 DCs，ELISA 方法分别检测各组细胞上清液中 IL －10、IL －12、转化生长因子β1（transformed growth factor －β1，TGFβ1）和血管内皮生长因子（vascular endothelial growth factor， VEGF）的表达情况。结果：共培养后，DCs 表达细胞因子 IL －12减少，DCs 和 K562细胞表达 TGFβ1增加， K562细胞表达 VEGF 增加。结论：共培养后，两种细胞在某些机制的相互作用下，DCs 表达 IL －12能力降低， DCs 和 K562细胞表达 TGFβ1的能力增高，K562细胞表达 VEGF 的能力提高，这可能是共培养后 DCs 免疫功能下降和血液肿瘤细胞逃脱机体免疫监视的细胞因子方面的原因。
目的：探討共培養條件下血液腫瘤 K562細胞和樹突狀細胞（dendritic cells，DCs）在錶達細胞因子方麵的相互作用情況，探索血液腫瘤髮生髮展過程中的免疫逃逸機製。方法：利用免疫磁珠試劑盒從人健康外週血中分離純化齣高純度的 CD14＋單覈細胞，一定濃度的巨噬細胞集落刺激因子（granlocyte －macrophage co-lomy stimulating factor，GM－CSF）和白介素4（interlenkins －4，IL －4）將單覈細胞誘導分化為未成熟 DCs（im-mature DCs，imDCs），再用一定濃度的腫瘤壞死因子（tumor necrosis factor －α，TNF －α）將 imDCs 誘導為成熟DCs（mature DCs，mDCs），分彆將 imDCs 和 mDCs 與 K562細胞在 Transwell 繫統中共培養48h，對照組為正常培養48h 的 DCs 和撤細胞因子培養48h 的 DCs，ELISA 方法分彆檢測各組細胞上清液中 IL －10、IL －12、轉化生長因子β1（transformed growth factor －β1，TGFβ1）和血管內皮生長因子（vascular endothelial growth factor， VEGF）的錶達情況。結果：共培養後，DCs 錶達細胞因子 IL －12減少，DCs 和 K562細胞錶達 TGFβ1增加， K562細胞錶達 VEGF 增加。結論：共培養後，兩種細胞在某些機製的相互作用下，DCs 錶達 IL －12能力降低， DCs 和 K562細胞錶達 TGFβ1的能力增高，K562細胞錶達 VEGF 的能力提高，這可能是共培養後 DCs 免疫功能下降和血液腫瘤細胞逃脫機體免疫鑑視的細胞因子方麵的原因。
목적：탐토공배양조건하혈액종류 K562세포화수돌상세포（dendritic cells，DCs）재표체세포인자방면적상호작용정황，탐색혈액종류발생발전과정중적면역도일궤제。방법：이용면역자주시제합종인건강외주혈중분리순화출고순도적 CD14＋단핵세포，일정농도적거서세포집락자격인자（granlocyte －macrophage co-lomy stimulating factor，GM－CSF）화백개소4（interlenkins －4，IL －4）장단핵세포유도분화위미성숙 DCs（im-mature DCs，imDCs），재용일정농도적종류배사인자（tumor necrosis factor －α，TNF －α）장 imDCs 유도위성숙DCs（mature DCs，mDCs），분별장 imDCs 화 mDCs 여 K562세포재 Transwell 계통중공배양48h，대조조위정상배양48h 적 DCs 화철세포인자배양48h 적 DCs，ELISA 방법분별검측각조세포상청액중 IL －10、IL －12、전화생장인자β1（transformed growth factor －β1，TGFβ1）화혈관내피생장인자（vascular endothelial growth factor， VEGF）적표체정황。결과：공배양후，DCs 표체세포인자 IL －12감소，DCs 화 K562세포표체 TGFβ1증가， K562세포표체 VEGF 증가。결론：공배양후，량충세포재모사궤제적상호작용하，DCs 표체 IL －12능력강저， DCs 화 K562세포표체 TGFβ1적능력증고，K562세포표체 VEGF 적능력제고，저가능시공배양후 DCs 면역공능하강화혈액종류세포도탈궤체면역감시적세포인자방면적원인。
Objective:To study the interactions of dendritic cells(DCs)and K562 when co -culture on cytokines expression,to investigate the mechanisms of leukemia immune escape.Methods:The CD14 + monocytes were purified from fresh peripheral blood of healthy human with immune magnetic beads,and then were cultured in the media con-taining certain concentration rhGM-CSF and IL -4 main to induce into immature DCs(imDCs),and mature DCs(mDCs)were developed by add certain concentration of rhTNF -αinto imDCs culture.The imDCs and mDCs were co -cultured with K562 cells in transwell chamber for 48h,those untreated cells including those cultured in normal medium for 48h and those cultured in medium without cytokines for 48h served and K562 cells cultured in 1640 for 48h as controls.The expression of IL -10,IL -12,transformed growth factor -β1 (TGFβ1 )and vascular endothelial growth factor(VEGF)were measured by ELISA.Results:After co -cultured,the expression of IL -12 of DCs were decreased(P <0.01),the expression of TGFβ1 of DCs and K562 cells were increased deeply (P <0.01)and the ex-pression of VEGF of K562 cells were increased obviously.Conclusion:After co -cultured,under the interaction of DCs and K562 cells in some mechanism,DCs ability of expressing IL -12 is reduced,DCs and K562 cells ability of expressing TGFβ1 were higher,and K562 cells ability of expressing VEGF were higher too,this could be the reason for DCs immune function damaged and K562 cells to escape immune surveillance on cytokine secretion.
