摘要
目的 :探讨肺癌患者外周血来源的树突状细胞 (dendriticcells,DC)生物学特性。方法 :从肺癌患者外周血分离获得单个核细胞 ,用细胞因子GM CSF (10 0 μg/L) ,IL 4 (5 0 μg/L)体外培养诱导。凋亡肿瘤细胞负载 2 4h后加入TNF α(10μg/L)或CD4 0激发型单抗于培养DC中 ,继续诱导 3～ 4d。分别测定DC的表型、DC摄取抗原能力 ;混合淋巴细胞反应(MLR)对T细胞增殖能力的测定 ;细胞计数和3 H TdR掺入观察DC对T细胞的激发和扩增效应 ,并与健康志愿者外周血来源的DC进行比较。结果 :患者外周血单个核细胞和正常人外周血单个核细胞来源的DC均高表达CD1α ,CD83,CD80 ,CD86和HLA DR等DC的相关抗原和共刺激分子 ;患者的未成熟DC能有效摄取FITC Dextran ,经TNF α或CD4 0激发型单抗激发诱导后 ,成为成熟和有功能的DC ,几乎完全失去对抗原的摄取能力 ,与健康人外周血来源DC组相比无明显差别 (P >0 .0 5 ) ;患者单个核细胞来源的DC在体外具有激发自体和同种异体外周血T细胞增殖的能力。结论 :肺癌患者的外周血来源单个核细胞可以诱导成为具有功能的成熟DC。
Objective: To study the biological characteristics of dendritic cells (DC) derived from peripheral blood of patients with lung cancer. Methods: Peripheral blood mononuclear cells (PBMC) were isolated from 11 lung cancer patients and healthy donors respectively, cultured with GM-CSF (100_g/L) and IL-4 (50_g/L). After pulsed with apoptotic lung cancer cells for 24 h, TNF-α(10_g/L)added or agonist CD40mAb (5 mg/L) for a further induction of 3～4 days. Phenotypes of DC were detected by FCM. The capacities for DC to uptake antigens were monitored at day 7 and day 11 detected by FITC-dextran uptake test. 3 H-thymidine incorporation test was used to measure the proliferative capability of T cells. Results: Both DC derived from PBMC of lung cancer patients and from PBMC of healthy donors showed high expressions of CD1α, CD80, CD83, CD86 and HLA-DR, which were known as associated markers of mature DC. The 7th day DC derived from lung cancer patients PBMC showed a strong capacity to uptake antigens. DC were induced by TNF-α or CD40mAb, then became mature. DC entirely lost their capacities of uptake antigens, while DC could stimulate autologous and allogenic T cells' proliferation. When matured DC co-incubated with autologous T cells for 48 h, activated T cells could be observed and the number of activated T cells was significantly increased ( P <0.05).Conclusion: The functional and mature DC could be induced from peripheral blood of lung cancer patients in vitro .
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
2003年第3期175-179,共5页
Chinese Journal of Cancer Biotherapy
基金
江苏省社会发展基金 (BS2 0 0 0 0 3 8)
江苏省医学重点人才基金 (RC2 0 0 2 0 3 2 )
