已入深夜,您辛苦了!由于当前在线用户较少,发布求助请尽量完整的填写文献信息,科研通机器人24小时在线,伴您度过漫漫科研夜!祝你早点完成任务,早点休息,好梦!

Epigenetic Profile of Treg-like Cells Induced By Mesenchymal Stem Cells in Vitro Resembles That of Natural Treg

FOXP3型 Treg细胞 间充质干细胞 免疫学 人口 过继性细胞移植 生物 表观遗传学 体外 骨髓 表型 外周血单个核细胞 免疫系统 癌症研究 白细胞介素2受体 医学 细胞生物学 T细胞 遗传学 基因 环境卫生
作者
Rita I. Azevedo,Ekaterina Minskaia,Cláudia L. da Silva,Ana Fernandes‐Platzgummer,Ana Vieira,Joaquim M. S. Cabral,João F. Lacerda
出处
期刊:Blood [American Society of Hematology]
卷期号:132 (Supplement 1): 2578-2578
标识
DOI:10.1182/blood-2018-99-112452
摘要

Abstract Regulatory T cells (Treg) are a key population in immune tolerance and their potential use in the treatment of chronic inflammatory diseases has been increasingly investigated. A major hurdle to adoptive Treg therapy in humans is the difficulty in obtaining sufficient numbers of clinical grade Treg. We and others have investigated the potential of mesenchymal stem cells (MSC) to induce and/or expand Treg in vitro. Our goal is to ascertain whether MSC co-culture supports the recruitment and expansion of Treg from a non-Treg starting population, which would allow for clinically relevant numbers to be reached, whilst bypassing the costly isolation of Treg under GMP conditions prior to expansion. We found that co-cultures of human peripheral blood mononuclear cells (PBMC) from healthy donors with allogeneic bone marrow-derived MSC specifically increase the absolute counts and frequency (4 and 6-fold, respectively) of Treg-like cells with a CD4+ CD25high Foxp3+ CD127low phenotype. We further observed that this increase in Treg numbers after MSC co-culture is mainly due to the induction of conventional CD4 T cells (Tcon) to acquire a Treg-like phenotype, rather than to Treg expansion. These data led us to investigate if these Treg-like cells induced by MSC also resemble Treg in terms of suppressive ability and epigenetic profile, which would inform on their functional potential and stability, two key features for their applicability in a clinical setting. We performed suppression assays using FACSorted Treg-like cells after co-culture of purified Tcon with MSC as suppressor cells and autologous fresh Tcon labelled with CFSE as responder cells. We compared the suppressive ability of Treg-like cells to that of Treg co-cultured with MSC, as well as of fresh Treg. We found that induced Treg-like cells isolated from Tcon:MSC co-cultures (P=0.0121), natural Treg isolated from Treg:MSC co-cultures (P=0.0025) and fresh Treg (P=0.0006) all significantly suppress Tcon proliferation (% divided cells at 1:1 Tcon:Treg ratio compared to Tcon alone, Mann Whitney test). We observed significantly higher levels of TGF-β in PBMS:MSC co-culture supernatants compared to that of PBMC cultured alone (P<0.05 on Day 10, P<0.01 on Day 14, two-way ANOVA), indicating a role for TGF-β in the induction of these Treg-like cells. TGF-β-induced Treg have been suggested to be less stable than natural Treg, likely due to epigenetic regulation. In particular, DNA demethylation has been shown to be a key factor in Treg stability. The potential therapeutic application of MSC-induced Treg-like cells would hinge on these cells being epigenetically stable. Hence, we investigated whether MSC-induced Treg-like cells are more similar to the original Tcon that they arose from or to natural Treg in terms of their DNA methylation profile. Most Treg epigenetic studies focus on the Treg-specific demethylated region (TSDR) of the FOXP3 gene and, given that FOXP3 is encoded on the X chromosome, opt to use male donors to bypass artefacts of X-chromosome inactivation. In order to avoid a gender biased analysis, we sought an alternative to TSDR. CAMTA1 (calmodulin-binding transciption activator 1) is a Ca2+-dependent calmodulin-binding transcription factor encoded on chromosome 1q3.6, which has been proposed as a potential molecular marker that distinguishes Treg from Tcon in both male and female donors. We assessed the methylation status of 13 CpG sites within the 470bp CAMTA1 intronic region 3 based on the coding DNA strand in Treg-like cells purified after Tcon:MSC co-cultures, as well as in fresh Treg and Tcon from the same donor. Fresh Tcon cells from one of the donors showed 100% methylation of CpGs 1-13, whereas fresh Treg and Treg-like cells from the same donor showed much lower levels of methylation (33.8% and 0%, respectively), with the highest differences in methylation observed in CpGs 2 and 11 (15% in fresh Treg; 0% in Treg-like cells; 100% in fresh Tcon). These two CpG positions, which have been previously demonstrated to be the ones distinguishing Treg from Tcon, were consistently more demethylated in fresh Treg and Treg-like cells than in fresh Tcon, with an average methylation levels of 5.8% for fresh Treg, 17% for Treg-like cells and 73% for fresh Tcon. Overall, our data demonstrate that MSC induce a subset of Tcon to acquire a Treg-like phenotype, which is accompanied by a suppressive ability and an epigenetic profile that resemble that of natural Treg. Disclosures No relevant conflicts of interest to declare.

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
1秒前
2秒前
someone发布了新的文献求助10
6秒前
6秒前
jyw完成签到,获得积分10
7秒前
标志的大寄完成签到,获得积分10
8秒前
随影相伴完成签到 ,获得积分10
8秒前
12秒前
kjding发布了新的文献求助10
15秒前
等风发布了新的文献求助10
17秒前
46464号发布了新的文献求助10
17秒前
zmx完成签到 ,获得积分10
18秒前
kekefefe发布了新的文献求助10
19秒前
Lan发布了新的文献求助10
19秒前
xx完成签到,获得积分10
22秒前
科小白完成签到 ,获得积分10
25秒前
天天快乐应助小笼包采纳,获得10
27秒前
28秒前
ok应助科研通管家采纳,获得10
29秒前
LaInh应助科研通管家采纳,获得10
29秒前
烂漫小蝴蝶完成签到,获得积分20
29秒前
科研通AI2S应助科研通管家采纳,获得10
29秒前
彭于晏应助科研通管家采纳,获得10
29秒前
清脆松应助科研通管家采纳,获得20
29秒前
充电宝应助科研通管家采纳,获得10
29秒前
华仔应助科研通管家采纳,获得10
29秒前
kid1912应助科研通管家采纳,获得10
29秒前
孙孙应助科研通管家采纳,获得10
29秒前
ok应助科研通管家采纳,获得10
29秒前
大模型应助科研通管家采纳,获得30
30秒前
LaInh应助科研通管家采纳,获得10
30秒前
30秒前
30秒前
香蕉觅云应助科研通管家采纳,获得10
30秒前
30秒前
31秒前
33秒前
33秒前
微风发布了新的文献求助20
33秒前
传奇3应助生物素采纳,获得10
33秒前
高分求助中
The ACS Guide to Scholarly Communication 2500
Sustainability in Tides Chemistry 2000
Pharmacogenomics: Applications to Patient Care, Third Edition 1000
Studien zur Ideengeschichte der Gesetzgebung 1000
TM 5-855-1(Fundamentals of protective design for conventional weapons) 1000
Threaded Harmony: A Sustainable Approach to Fashion 810
《粉体与多孔固体材料的吸附原理、方法及应用》(需要中文翻译版,化学工业出版社,陈建,周力,王奋英等译) 500
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3084988
求助须知:如何正确求助?哪些是违规求助? 2738035
关于积分的说明 7547906
捐赠科研通 2387624
什么是DOI,文献DOI怎么找? 1266055
科研通“疑难数据库(出版商)”最低求助积分说明 613267
版权声明 598450