中华整形外科杂志
中華整形外科雜誌
중화정형외과잡지
CHINESE JOURNAL OF PLASTIC SURGERY
2015年
1期
43-48
,共6页
丁刚%魏立梅%孙伟元%张丽
丁剛%魏立梅%孫偉元%張麗
정강%위립매%손위원%장려
扁桃体%间质干细胞%免疫
扁桃體%間質榦細胞%免疫
편도체%간질간세포%면역
Tonsil%Mesenchymal stem cells%Immunity
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疫抑制特性,有望进行同种异体移植.
目的 探討扁桃體間充質榦細胞(tonsil mesenchymal stem cells,TMSCs)的免疫學特性及機製.方法 取慢性扁桃體炎患兒的扁桃體組織,分離、培養TMSCs,通過流式細胞術檢測HLA-Ⅰ、HLA-Ⅱ、CD80、CD86等免疫分子的錶達情況.以牙週膜榦細胞作為對照,觀察TMSCs能否引起同種異體外週血單箇覈細胞(peripheral blood mononuclear cells,PBMCs)的增殖,以及TMSCs對混閤淋巴細胞反應(mixed lymphocyte reaction,MLR)和植物血凝素(phytohemagglutinin,PHA)引起的淋巴細胞增殖的影響.建立TMSCs+PHA+異體PBMCs、TMSCs+MLR的培養體繫,測定細胞上清液中的犬尿氨痠濃度.在上述反應體繫進行中和實驗,觀察被TMSCs抑製瞭的淋巴細胞重新髮生增殖的情況.每箇實驗重複3次,每組6箇佯本.統計學方法採用方差分析,P ＜0.05為差異有統計學意義.結果 TMSCs錶達HLA-Ⅰ,但不錶達HLA-Ⅱ和共刺激分子CD80、CD86.TMSCs與異體PBMCs共培養5d後,刺激指數為1.38±0.26,而單純PBMCs培養5d後的刺激指數為1.22±0.28,2組差異無統計學意義(P＞0.05),證實TMSCs不會引起異體PBMCs增殖.TMSCs與異體PBMCs、PHA共培養5d後,刺激指數分彆為1.49±0.29(TMSCs∶ PBMCs為0.5∶1)和1.23±0.22(TMSCs∶ PBMCs為1∶1),而PBMCs+PHA組培養5d後的刺激指數為4.60±0.81,2組之間的差異均有統計學意義(P＜0.05),說明TMSCs能夠抑製PHA引起的淋巴細胞增殖.TMSCs與MLR共培養5d後,刺激指數分彆為1.29±0.23(TMSCs∶ PBMCs為0.5∶1)和1.26±0.27(TMSCs∶ PBMCs為1∶1),而MLR培養5d後的刺激指數為3.04±0.66,2組之間的差異均有統計學意義(P＜0.05),說明TMSCs能夠抑製MLR引起的淋巴細胞增殖.在TMSCs+PHA+異體PBMCs、TMSCs+MLR的培養體繫中,犬尿氨痠濃度顯著升高,分彆為(26.0±2.3) μmol/L和(23.5±4.5)μmol/L.中和實驗髮現,1-甲基-L-色氨痠基本恢複瞭被TMSCs抑製的淋巴細胞增殖.結論 TMSCs具有低免疫原性和免疫抑製特性,有望進行同種異體移植.
목적 탐토편도체간충질간세포(tonsil mesenchymal stem cells,TMSCs)적면역학특성급궤제.방법 취만성편도체염환인적편도체조직,분리、배양TMSCs,통과류식세포술검측HLA-Ⅰ、HLA-Ⅱ、CD80、CD86등면역분자적표체정황.이아주막간세포작위대조,관찰TMSCs능부인기동충이체외주혈단개핵세포(peripheral blood mononuclear cells,PBMCs)적증식,이급TMSCs대혼합림파세포반응(mixed lymphocyte reaction,MLR)화식물혈응소(phytohemagglutinin,PHA)인기적림파세포증식적영향.건립TMSCs+PHA+이체PBMCs、TMSCs+MLR적배양체계,측정세포상청액중적견뇨안산농도.재상술반응체계진행중화실험,관찰피TMSCs억제료적림파세포중신발생증식적정황.매개실험중복3차,매조6개양본.통계학방법채용방차분석,P ＜0.05위차이유통계학의의.결과 TMSCs표체HLA-Ⅰ,단불표체HLA-Ⅱ화공자격분자CD80、CD86.TMSCs여이체PBMCs공배양5d후,자격지수위1.38±0.26,이단순PBMCs배양5d후적자격지수위1.22±0.28,2조차이무통계학의의(P＞0.05),증실TMSCs불회인기이체PBMCs증식.TMSCs여이체PBMCs、PHA공배양5d후,자격지수분별위1.49±0.29(TMSCs∶ PBMCs위0.5∶1)화1.23±0.22(TMSCs∶ PBMCs위1∶1),이PBMCs+PHA조배양5d후적자격지수위4.60±0.81,2조지간적차이균유통계학의의(P＜0.05),설명TMSCs능구억제PHA인기적림파세포증식.TMSCs여MLR공배양5d후,자격지수분별위1.29±0.23(TMSCs∶ PBMCs위0.5∶1)화1.26±0.27(TMSCs∶ PBMCs위1∶1),이MLR배양5d후적자격지수위3.04±0.66,2조지간적차이균유통계학의의(P＜0.05),설명TMSCs능구억제MLR인기적림파세포증식.재TMSCs+PHA+이체PBMCs、TMSCs+MLR적배양체계중,견뇨안산농도현저승고,분별위(26.0±2.3) μmol/L화(23.5±4.5)μmol/L.중화실험발현,1-갑기-L-색안산기본회복료피TMSCs억제적림파세포증식.결론 TMSCs구유저면역원성화면역억제특성,유망진행동충이체이식.
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 ±0.27,respectively,however,the stimulation index in the group of MLR was 3.04 ± 0.66,and the difference between the two groups had a statistical significance (P ＜ 0.05),demonstrating that TMSCs could suppress MLR-induced PBMCs proliferation.The levels of kynurenine were (26.0 ± 2.3) μmol/L and (23.5 ± 4.5) μmol/L in the culture of TMSCs + PBMCs + PHA and TMSCs + MLR,respectively,thus elevating significantly.After adding of 1-methyl-L-tryptophan,TMSCs-mediated-proliferation suppression of PBMCs restored to normal levels.Conclusion TMSCs possess low immunogenecity and immunosuppressive function,may be used in allogeneic transplantation.
