细胞因子诱导杀伤细胞/自然杀伤细胞培养过程中CD16及CD11a的表达变化  

Changes of CD16 and CD11a expression in cytokine-induced killer cells/natural killer cells during cultivation

作　　者：黎阳[1] 吴燕峰[2] 曾颖[3] 方建培[1] 魏菁[4] 周敦华[1] 郭海霞[1] 黄绍良[1] 林永潮[1] 

机构地区：[1]中山大学附属第二医院儿科,广东省广州市510120 [2]中山大学附属第二医院干细胞研究中心,广东省广州市510120 [3]中山大学附属第二医院整形外科,广东省广州市510120 [4]中山大学附属第二医院医学研究中心,广东省广州市510120

出　　处：《中国组织工程研究与临床康复》2009年第5期882-886,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30500469);广州科学仪器协作共用网基金资助项目(2006034);中山大学北校区学生科研基金资助项目(2006031)~~

摘　　要：背景:由于细胞因子诱导杀伤细胞、自然杀伤细胞上表达的CD16和CD11a均与其介导的抗体依赖性细胞介导的细胞毒作用和直接接触杀伤的抗肿瘤效应密切相关,因此了解两种细胞扩增过程中这两种分子表达的强弱,对根据免疫效应细胞的最终用途决定两种细胞的最佳收获时机具有重要意义。目的:了解细胞因子诱导杀伤细胞和自然杀伤细胞在培养过程中的CD16和CD11a表达的变化规律。设计、时间及地点:细胞移植免疫学动态观察,于2006-09/2008-03在中山大学附属第二医院完成。材料:脐血来自本院健康足月妊娠产妇。方法:采用Ficoll-Hypaque法分离脐血单个核细胞,接种于含体积分数为0.15胎牛血清的IMDM,双抗生素青、链霉素各1×105U/L24孔培养板中,在培养体系中加入白细胞介素2、白细胞介素7、白细胞介素15、干细胞因子及FLT3L制备细胞因子诱导杀伤细胞和自然杀伤细胞,每隔3天半量换液及全量补充上述细胞因子。主要观察指标:流式细胞仪检测CD3+CD56+细胞因子诱导杀伤细胞以及CD3-CD56+自然杀伤细胞在4周培养过程中CD16和CD11a表达的变化。结果:CD16在细胞因子诱导杀伤细胞、自然杀伤细胞上的表达随着培养时间的延长逐渐增加,在两类细胞上均在培养的第4周达到最高峰,分别为(55.99±3.90)%和(7.86±1.66)%,但CD16在自然杀伤细胞上表达比例较低,整个培养过程中自然杀伤细胞上CD16的均数表达没有超过8%。CD11a在细胞因子诱导杀伤细胞培养的第3周,自然杀伤细胞培养的第2周达到最高峰,分别为(49.32±6.32)%和(82.31±11.33)%,之后逐渐出现下降,到培养第4周几乎消失。结论:CD16和CD11a在细胞因子诱导杀伤细胞和自然杀伤细胞上的表达随培养时间呈动态变化,使用单克隆抗体联合细胞因子诱导杀伤细胞介导的抗体依赖性细胞介导的细胞毒作用效应时,细胞的收获期可在细胞培养的第4周,�BACKGROUND： Expression of CD16 and CD1 la on cytokine-induced killer （CIK） cells and natural killer （NK） cells plays an important role in the potential of antibody dependent cell-mediated cytotoxicity or directed lysis activities against tumor cells, it has important significance in selecting the proper harvest time of CIK and NK cells by studying on the change law of expression of CD16 and CD1 la on CIK cells and NK cells in the process of cultivation. OBJECTIVE： To explore the change law of of CD16 and CD11 a expression on cytokine-induced killer cells/natural killer cells in the process of cultivation. DESIGN, TIME AND SETTING： The immunology dynamic observation on cell transplantation was performed at the Second Affiliated Hospital of Sun Yat-sen University from September 2006 to March 2008. MATERIAL： The umbilical cord blood was collected from normal parturients in hospital. METHODS： Cord blood mononuclear cells were isolated by Ficoll-Hypaque density gradient centrifugation, and cultured in 24 well culture plate of IMDM containing 15% fetal calf serum, penicillin 1 × 10^5 U/L, and streptomycin 1 × 10^5 U/L. To generate CIK and NK cells, Interleukin-2, Interleukin-7, Interleukin-15, stem cell factor and FLT3L were added periodically, and the cells were feeded every three days. MAIN OUTCOME MEASURES： To explore the change laws of CD16 and CD1 la by detecting CD3^＋CD56^＋ CIK cells and CD3CD56^＋ NK cells during four-week of culture with flow cytometry. RESULTS： The expression of CD16 on CIK cells increased gradually according to the cultural time and reached the peak value at the end of the fourth week. The CD16 expressed on NK cells was in a relatively low and stable manner during the process. Along with prolongation of the cultural time, the expression of CD16 on CIK and NK cells increased gradually, and both reached the peak value at （55.99±3.90）% and （7.86±1.66）%, respectively after four weeks cultivation. The expression of CD16 was relatively lower on NK

关 键 词：细胞因子诱导杀伤细胞 自然杀伤细胞 CD16 CD11A 过继免疫治疗 

分 类 号：R617[医药卫生—外科学]