脑源性神经营养因子增强间充质干细胞抑制滤泡辅助性T细胞的作用

目的探讨脑源性神经营养因子(BDNF)增强间充质干细胞(MSC)抑制滤泡辅助性T细胞(Tfh细胞)的作用及机制。方法ELISA法检测MSC培养上清中吲哚胺2,3-二加氧酶(IDO)、IL.10、TGF—B和IL-2l的含量;采集健康志愿者的外周血标本,采用人淋巴细胞分离液分离外周血中的淋巴细胞;采用Transwell小室进行MSC和淋巴细胞共培养,流式细胞术检测CD4+CXCR5+Tfh细胞及其亚群的比例。结果@BDNF组(BDNF刺激的MSC)培养上清IL-10、TGF-β、IDO浓度均高于对照组(加入等体积磷酸盐缓冲液)[IL-10:(42.1±64.4)ng/ml对(19.3±2.1)n...

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Bibliographic Details
Published in中华血液学杂志 Vol. 39; no. 1; pp. 37 - 40
Main Author 杨赛男;蒲欣;向莎丽;陈洁平;裴莉
Format Journal Article
LanguageChinese
Published 陆军军医大学附属西南医院血液科,重庆,400038 2018
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ISSN0253-2727
DOI10.3760/cma.j.issn.0253-2727.2018.01.008

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Summary:目的探讨脑源性神经营养因子(BDNF)增强间充质干细胞(MSC)抑制滤泡辅助性T细胞(Tfh细胞)的作用及机制。方法ELISA法检测MSC培养上清中吲哚胺2,3-二加氧酶(IDO)、IL.10、TGF—B和IL-2l的含量;采集健康志愿者的外周血标本,采用人淋巴细胞分离液分离外周血中的淋巴细胞;采用Transwell小室进行MSC和淋巴细胞共培养,流式细胞术检测CD4+CXCR5+Tfh细胞及其亚群的比例。结果@BDNF组(BDNF刺激的MSC)培养上清IL-10、TGF-β、IDO浓度均高于对照组(加入等体积磷酸盐缓冲液)[IL-10:(42.1±64.4)ng/ml对(19.3±2.1)ng/ml,t=4.761,P=0.009;TGF—β:(13.9±1.7)ng/ml对(5.3±0.6)ng/ml,t=5.129,P=0.008;IDO:(441.3±56.9)ng/ml对(226.7±37.6)ng/ml,t=3.130,P=O.035];@BDNF组(淋巴细胞与BDNF刺激的MSC共培养)与MSC组(淋巴细胞与MSC共培养)比较:CD4+CXCR5+Tfh细胞比例降低[(3.37+0.21)%对(6.51+0.27)%,t=9.353,P〈0.001],CD4+CXCR5+CXCR3+CCR6-Tfhl细胞比例升高[(41.14±2.04)%对(26.72±2.57)%,t=4.383,P=0.012],CD4+CXCR5+CXCR3CCR6-Tfh2细胞和CD4+CXCR5+CXCR3-CCR6+Tfhl7细胞比例降低[Tfh2:(30.16:L5.38)%对(43.26+4.11)%,t:4.426,P=0.012;Tfhl7:(15.61±1.52)%对(22.32±0.72)%,t=4.202,P:0.014],CD4+CXCR5+Foxp3+Tfr细胞比例升高[(4.95±0.22)%对(2.32±0.16)%,t=10.241,P〈0.001],淋巴细胞培养上清中IL-21浓度降低[(0.28±0.03)ng/ml对(0.85±0.08)ng/ml,t=6.675,P=0.003]。结论BDNF能够增强MSC抑制Tfh细胞的作用,机制是抑制淋巴细胞中Tfh细胞比例升高及其向Tfh2和Tfhl7亚群的分化。
Bibliography:Objective To investigate the effect of brain derived neurotrophic factor (BDNF) on mesenchymal stem cells (MSC) inhibiting follicular helper T cells (Tfh cells). Methods The contents of indoleamine 2,3-dioxygenase (IDO), IL-10, TGF-β and IL-21 in MSC culture supematant were detected by ELISA; The peripheral blood of healthy volunteers were collected, and lymphocyte in peripheral blood was separated by human lymphocyte separation solution; Co-cultures of MSC and lymphocyte were performed by Transwell chamber, and the proportion of CD4+CXCR5 + Tfh cells and their subtypes were detected by flow cytometry. Results (1)The concentrations of IL- 10, TGF-β, and IDO in the supernatant of BDNF group (BDNF-stimulated MSC) were higher than those of the control ones (adding PBS with the same volume) [IL-10: (42.1±4.4) ng/ml vs (19.3±2.1) ng/ml, t=4.761, P= 0.009; TGF-β: (13.9±1.7) ng/ml vs (5.3±0.6) ng/ml, t= 5.129, P= 0.008; IDO: (441.3±56.9) ng/ml vs (226.7±37.6) ng/ml, t= 3.130, P= 0.035]; (2)The comparisons between BD
ISSN:0253-2727
DOI:10.3760/cma.j.issn.0253-2727.2018.01.008