大肠癌患者外周血来源树突状细胞对T细胞活化作用的研究  

Effects of peripheral blood derived dendritic cells on T cell activity in colorectal cancer patients.

作　　者：王建鑫[1] 陈文明[1] 刘晋玮[1] 齐曼[1] 郝建宇[1] 关玉盘[1] 王世鑫[1] 

机构地区：[1]首都医科大学附属北京朝阳医院消化内科,北京100020

出　　处：《中国实用内科杂志：临床前沿版》2006年第9期1414-1417,共4页

基　　金：北京市科技新星项目(954812300)

摘　　要：目的比较无血清培养液AIM-V和RPMI1640+胎牛血清(FBS)培养液培养大肠癌患者外周血来源树突状细胞(DCs)的形态和免疫表型,检测DCs对T细胞的活化作用。方法对2003年3月至2004年10月首都医科大学附属北京朝阳医院消化内科收治的10例大肠癌患者,采集其外周血分离单核细胞,分别使用2种细胞培养液进行体外培养。观察细胞形态,检测细胞表面CD1a,CD80,CD86,CD83,CD14及HLA-DR的表达。再用丝裂霉素作用后的DCs作为刺激细胞,按照刺激细胞反应细胞(S∶R)1∶1,1∶10,1∶20,1∶40,1∶80的比例进行同种异体混合淋巴细胞反应(MLR),检测DCs刺激同种异体T细胞增殖的能力。结果大肠癌患者外周血单核细胞经2种细胞培养液培养后都可以诱导出DCs。细胞外型均呈毛刺状。细胞均表达CD1a、CD80、CD86和人类白细胞抗原(HLA-DR),不表达CD14、CD83,为不成熟DCs的特点。两组DCs细胞表型表达差异无显著性(P>0.05)。按S∶R不同比例进行同种异体MRL所得刺激指数分别为2.7,4.1,3.1,2.5,1.6,表明树突状细胞(DCs)在不同比例下都刺激T细胞增殖。结论使用AIM-V无血清培养液和传统的含FBS细胞培养液均可成功诱导培养大肠癌患者外周血来源的DCs,但使用AIM-V可以降低含血清培养液应用人体后引起的过敏反应等风险,更适合DCs的临床使用,可以替代含血清培养液。MLR结果显示,DCs在各种S∶R比例下均有刺激T细胞增殖的能力,但S∶R比为1∶10时最佳。Objective To compare the morphology and immunophenotype of dendritic cells （DCs） from peripheral blood in the patients with colorectal cancer cultivated in AIM - V serum - free media or in RPMI1640 supplemented with fetal bovine serum （FBS）, and to detect the proliferation activity of allogenic T cells stimulated by DCs. Methods From Mar. 2003 to Oct. 2004, monocytes from the colorectal cancer patients were isolated in vitro, and then were cultivated in two different media, respectively. DCs were harvested after 7 days culture. The morphology of DCs in two different media was photographed by inverted microscope. The DCs immunophenotype of CD1a, CD14, CD80, CD86, CD83 and HLA - DR were detected by flow cytometry. The proliferation activity of allogenic T cells （reactor） stimulated by DCs （stimulator）, which had been cultivated in mitomycin -contained media, was detected by allogeneic mixed lymphocyte reaction （MLR） according to stimulator -to -reactor ratio 1： 1, 1： 10, 1：20, 1：40 and 1： 80. Results DCs could be induced in two different media. There was no obvious difference of DCs morphology, which was in the shape of small spinose. The DCs met the immunophenotypical characteristics of immature DCs, and expressed CD1a, CD80, CD86 and HLA - DR, not expressed CD14 and CD83 were not expressed. There was no significant statistical difference of immunophenotypical expression of the two media derived DCs （P 〉 0. 05）. The stimulative index （SI） tested by MTT in MLR at stimulator - to - reactor cell ratio 1：1, 1：10, 1：20, 1：40 and 1：80 was 2. 7, 4.1, 3.1, 2.5 and 1.6,respectively. The proliferation of T cells stimulated by DCs could be detected at all stimulator - to - reactor ratio. Conclusion DCs could be successfully induced from peripheral blood monocyte in the patients with colorectal cancer by AIM - V serum - free media or by traditional FBS - contained media. AIM - V is more suitable for clinical immunotherapy because it can avoid the risk of allergic reaction for

关 键 词：大肠癌 树突状细胞 T细胞增殖 

分 类 号：R5[医药卫生—内科学]