扁桃体间充质干细胞免疫学特性的初步研究  

The immunological characteristics of tonsil mesenchymal stem cells

作　　者：丁刚[1] 魏立梅[1] 孙伟元[2] 张丽[1] 

机构地区：[1]潍坊医学院附属益都中心医院口腔科,青州262500 [2]潍坊医学院附属益都中心医院耳鼻咽喉科,青州262500

出　　处：《中华整形外科杂志》2015年第1期43-48,共6页Chinese Journal of Plastic Surgery

基　　金：国家自然科学基金（81070799）;山东省优秀中青年科学家科研奖励基金（BS2010SW033）

摘　　要：目的 探讨扁桃体间充质干细胞（tonsil mesenchymal stem cells,TMSCs）的免疫学特性及机制.方法 取慢性扁桃体炎患儿的扁桃体组织,分离、培养TMSCs,通过流式细胞术检测HLA-Ⅰ、HLA-Ⅱ、CD80、CD86等免疫分子的表达情况.以牙周膜干细胞作为对照,观察TMSCs能否引起同种异体外周血单个核细胞（peripheral blood mononuclear cells,PBMCs）的增殖,以及TMSCs对混合淋巴细胞反应（mixed lymphocyte reaction,MLR）和植物血凝素（phytohemagglutinin,PHA）引起的淋巴细胞增殖的影响.建立TMSCs＋PHA＋异体PBMCs、TMSCs＋MLR的培养体系,测定细胞上清液中的犬尿氨酸浓度.在上述反应体系进行中和实验,观察被TMSCs抑制了的淋巴细胞重新发生增殖的情况.每个实验重复3次,每组6个佯本.统计学方法采用方差分析,P ＜0.05为差异有统计学意义.结果 TMSCs表达HLA-Ⅰ,但不表达HLA-Ⅱ和共刺激分子CD80、CD86.TMSCs与异体PBMCs共培养5d后,刺激指数为1.38±0.26,而单纯PBMCs培养5d后的刺激指数为1.22±0.28,2组差异无统计学意义（P＞0.05）,证实TMSCs不会引起异体PBMCs增殖.TMSCs与异体PBMCs、PHA共培养5d后,刺激指数分别为1.49±0.29（TMSCs∶ PBMCs为0.5∶1）和1.23±0.22（TMSCs∶ PBMCs为1∶1）,而PBMCs＋PHA组培养5d后的刺激指数为4.60±0.81,2组之间的差异均有统计学意义（P＜0.05）,说明TMSCs能够抑制PHA引起的淋巴细胞增殖.TMSCs与MLR共培养5d后,刺激指数分别为1.29±0.23（TMSCs∶ PBMCs为0.5∶1）和1.26±0.27（TMSCs∶ PBMCs为1∶1）,而MLR培养5d后的刺激指数为3.04±0.66,2组之间的差异均有统计学意义（P＜0.05）,说明TMSCs能够抑制MLR引起的淋巴细胞增殖.在TMSCs＋PHA＋异体PBMCs、TMSCs＋MLR的培养体系中,犬尿氨酸浓度显著升高,分别为（26.0±2.3） μmol/L和（23.5±4.5）μmol/L.中和实验发现,1-甲基-L-色氨酸基本恢复了被TMSCs抑制的淋巴细胞增殖.结论 TMSCs具有低免疫原性和免�Objective To investigate the immunological characteristics of human tonsil mesenchymal stem cells （TMSCs）.Methods Human tonsil tissues were obtained from the children patients with chronic tonsillitis.TMSCs were separated,cultured,and were detected the expression profiles of HLA-Ⅰ,HLA-Ⅱ,CD80,CD86 by flow cytometry.The measurement of immunogenicity,the effect on phytohemagglutinin（PHA） induced peripheral blood mononuclear cell （PBMCs） proliferation and mixed lymphocytes reaction （MLR） were performed to identify the immunological characteristics of TMSCs.The co-cultures of TMSCs ＋ PBMCs ＋ PHA and TMSCs ＋ MLR were established,respectively,and the concentration of kynurenine,which is the metabolin of indoleamine 2,3-dioxygenase,in the culture supernatant were examined.Then we added 1-methyl-L-tryptophan into the co-culture of TMSCs ＋ PBMCs ＋ PHA and TMSCs ＋ MLR,respectively,and tested the proliferation of PBMCs.Each experiment was repeated three times,and there were six samples in each group.Statistical significance was assessed by analysis of variance （ANOVA）,and a P value less than 0.05 was considered statistically significant.Results TMSCs expressed HLA-Ⅰ,were negative for HLA-Ⅱ and co-stimulatory molecules CD80 and CD86.The stimulation index in the group of TMSCs ＋ allogeneic PBMCs was 1.38 ± 0.26,whereas the stimulation index in the group of allogeneic PBMCs was 1.22 ± 0.28,and there was no significant difference between the two groups （P 〉 0.05）,indicating that TMSCs could not initiate the proliferation of allogeneic PBMCs.The stimulation indexes in the group of TMSCs ＋ allogeneic PBMCs ＋ PHA were 1.49 ± 0.29 and 1.23 ± 0.22,respectively,whereas the stimulation index in the group of allogeneic PBMCs ＋ PHA was 4.60 ± 0.81,and the difference between the two groups had a statistical significance（P 〈 0.05）,suggesting that TMSCs could inhibit PHA-induced PBMCs proliferation.The stimulation indexes in the group of TMSCs ＋ MLR were 1.29 ±0.23 and 1.26

关 键 词：扁桃体 间质干细胞 免疫 

分 类 号：R392[医药卫生—免疫学]