OP0260 HEALTHY ENTHESEAL IMMUNOREGULATION MAINLY MEDIATED BY CD39/CD73 ADENOSINE PATHWAYS

腺苷 医学 计算机科学 内分泌学
作者
A. Altaie,Davide Simone,Heather Owston,N. McDermott,Stephen N. Sansom,Dennis McGonagle
出处
期刊:Annals of the Rheumatic Diseases [BMJ]
标识
DOI:10.1136/annrheumdis-2024-eular.4945
摘要

Background:

The enthesis is a pivotal target tissue in spondyloarthritis (SpA), marked by inflammation in both the enthesis soft tissue (EST) and the contiguous anchoring peri-entheseal bone (PEB) [1]. The healthy human spinal enthesis harbors resident mesenchymal stem cells (MSCs), myeloid cells, innate immune cell populations, while notably lacking FOXP3+ regulatory T cells (Treg) [2, 3]. In a previous investigation, we posited that MSCs within the enthesis, as opposed to regulatory T cells, are pivotal contributors to the observed physiological immunomodulation in this tissue [4].

Objectives:

This study seeks to elucidate whether the immunosuppressive pathway mediated by CD39/CD37 ectonucleotidase enzymes and adenosine is responsible for the immunosuppressive effects exerted by entheseal MSCs.

Methods:

Samples from healthy spinous processes and interspinous ligaments were divided into EST and PEB regions, enzymatically digested, and analyzed via flow cytometry. Cells were expanded to passage 3-5 (n=5) for co-culture experiments. CD4+ CD25- cells (T cells) were isolated from a single healthy donor and induced to proliferate using CD3/CD28 stimulation. Co-culture of MSCs and T cells was conducted at ratios of 1:1, 1:2, 1:4, and 1:8 for 5 days. The Flowjo software was employed to calculate the T cell proliferation index. Treg-related markers, including CD73 and CD39, were assessed cytometrically in PEB and EST. CD39 and CD73 were blocked using POM-1 and AOPCP, followed by co-culture with stimulated T cells for 5 days. Total adenosine production was measured using an Adenosine Assay Kit.

Results:

Activated T cells significantly upregulated CD39/CD73 on MSCs (EST and PEB) surfaces (P<0.001). In vivo uncultured EST and PEB MSCs exhibited the expression of CD39/CD73, affirming the translational relevance of this study. Co-culture of MSCs EST and PEB with activated T cells at a 1:1 ratio resulted in a significant suppression of T cell proliferation by 79% and 89%, respectively, compared to activated T cells alone. RNA-seq analysis of MSCs post co-culture indicated the upregulation of 390 genes and downregulation of 306 genes in EST MSCs, while 119 genes were upregulated, and 90 genes were downregulated in PEB MSCs (P<0.05). This analysis revealed the upregulation of shared immunosuppression pathways with Treg cells, including ENTPD3 (CD39), NT5E (CD73), CTLA-4, IL10, CCR4, CCR8, Foxp3, PTGER4, IL6, IDO1, PDL-1. Along with the upregulation of osteogenesis markers (CASR, APL) and downregulation of adipogenic markers (PPARG, SFRP4) and chondrogenic markers (GDF6, COL1A2). Simultaneously, IL17A and IL17F were upregulated and TGFβ downregulated. Blockage of CD39, CD73, and dual blockage increased T cell proliferation at 1:1 ratio with EST MSCs by 17.4%, 23.1%, and 52.2%, and with PEB MSCs by 32.1%, 41.9%, and 54.7%, respectively. This increase in T cell proliferation occurred concurrently with the inhibition of adenosine production. RNA-seq analysis of MSCs after adenosine inhibition via dual blockage of CD39/CD73 influenced immunosuppression markers shared with Treg cell pathways, resulting in the downregulation of CTLA-4, IL10, FOXP3, HLA-G, IL6, and IDO1, while CCR4, CCR8, and Foxp3 worked independently of adenosine production. Adenosine inhibition suppressed osteogenesis markers (CASR, APL) in EST and PEB MSCs, while upregulating adipogenic markers (PPARG, SFRP4).

Conclusion:

This study provides evidence that the MSCs derived from enthesal tissues possess an intrinsic immunomodulation capacity regulated by inflammation. These findings exert a significant influence on T cell activity, supporting the involvement of the CD39/73 adenosine pathway.

REFERENCES:

[1] Benjamin, M. et al., Scand J Med Sci Sports, 2009. 19(4): p. 520-7. [2] Watad, A., et al.,. Annals of the Rheumatic Diseases, 2020. 79(8): p. 1044. [3] Russell, T., et al.,. Cells, 2021. 10(2).4. [4] Altaie, A., et al., op0105, Annals of the Rheumatic Diseases, 2023. 82(Suppl 1): p. 70-71.

Acknowledgements:

NIL.

Disclosure of Interests:

None declared.

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

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
跳脚的虾完成签到 ,获得积分10
3秒前
JSEILWQ完成签到 ,获得积分10
7秒前
innocent完成签到,获得积分10
10秒前
彩色靖儿完成签到 ,获得积分10
15秒前
岚12完成签到 ,获得积分10
17秒前
JUAN完成签到,获得积分10
21秒前
陶醉的翠霜完成签到 ,获得积分10
22秒前
Novice6354完成签到 ,获得积分10
24秒前
心灵美绝施完成签到 ,获得积分10
28秒前
大雪完成签到 ,获得积分10
34秒前
呆橘完成签到 ,获得积分10
35秒前
22222应助yhm7426采纳,获得50
35秒前
36秒前
life的半边天完成签到 ,获得积分10
42秒前
灵巧的十八完成签到 ,获得积分10
43秒前
空曲完成签到 ,获得积分10
48秒前
小文子完成签到 ,获得积分10
49秒前
可可可爱完成签到 ,获得积分10
50秒前
鹏826完成签到 ,获得积分0
52秒前
zjq完成签到 ,获得积分10
52秒前
李爱国应助故意的鼠标采纳,获得10
53秒前
牧绯完成签到,获得积分10
56秒前
57秒前
Superman完成签到 ,获得积分10
1分钟前
乘风破浪完成签到 ,获得积分10
1分钟前
贝贝完成签到 ,获得积分10
1分钟前
Arctic完成签到,获得积分10
1分钟前
调皮蛋完成签到,获得积分10
1分钟前
reset完成签到 ,获得积分10
1分钟前
suodeheng完成签到,获得积分20
1分钟前
SY15732023811完成签到 ,获得积分10
1分钟前
www完成签到 ,获得积分10
1分钟前
leo完成签到 ,获得积分10
1分钟前
天天快乐应助帅气的宛凝采纳,获得10
1分钟前
RandyChen完成签到,获得积分10
1分钟前
kehe!完成签到 ,获得积分0
1分钟前
Nemo完成签到 ,获得积分10
1分钟前
1分钟前
1分钟前
英勇海完成签到 ,获得积分10
1分钟前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
Cognitive Neuroscience: The Biology of the Mind 1000
Technical Brochure TB 814: LPIT applications in HV gas insulated switchgear 1000
Immigrant Incorporation in East Asian Democracies 600
Nucleophilic substitution in azasydnone-modified dinitroanisoles 500
不知道标题是什么 500
A Preliminary Study on Correlation Between Independent Components of Facial Thermal Images and Subjective Assessment of Chronic Stress 500
热门求助领域 (近24小时)
化学 材料科学 医学 生物 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 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3968559
求助须知:如何正确求助?哪些是违规求助? 3513391
关于积分的说明 11167370
捐赠科研通 3248808
什么是DOI,文献DOI怎么找? 1794465
邀请新用户注册赠送积分活动 875116
科研通“疑难数据库(出版商)”最低求助积分说明 804664