A Novel Human OX40L-CAR-Treg Specifically Targets Activated Antigen-Presenting Cells and Effectively Controls T Cell Alloreactivity and Xeno-GvHD

FOXP3型 免疫学 白细胞介素2受体 抗原 嵌合抗原受体 白细胞介素-7受体 人类白细胞抗原 过继性细胞移植 CD28 移植 医学 免疫系统 癌症研究 生物 T细胞 CD8型 外科
作者
Xianliang Rui,Francesca Alvarez Calderon,Ulrike Gerdemann,Connor McGuckin,Lorenzo Cagnin,Bruce R. Blazar,Victor Tkachev,Leslie S. Kean
出处
期刊:Blood [Elsevier BV]
卷期号:140 (Supplement 1): 871-872 被引量:1
标识
DOI:10.1182/blood-2022-165745
摘要

Background: Graft-versus-host disease (GvHD) remains the leading cause of non-relapse mortality after allogeneic hematopoietic stem cell transplantation (HCT). Adoptive transfer of regulatory T cells (Tregs) has demonstrated promising results in controlling GvHD in both animal models and phase I/II clinical trials. However, strategies to increase the stability of ex-vivo expanded Tregs, and to optimize their potency, are needed. Recent efforts to accomplish these goals have focused on engineering Chimeric Antigen Receptor (CAR) Tregs, with the first attempts using CARs targeted against disparate HLA antigens. However, Tregs targeting HLA have several limitations, including the downregulation of these antigens that accompanies some diseases, as well as the lack of utility of this approach in the HLA-matched setting. In this study, we tested a novel GvHD prophylaxis approach consisting of infusing donor Tregs engineered to express an anti-OX40L CAR that, upon engagement with activated OX40L+ host antigen-presenting cells (APCs), increases their suppressor potency to dampen APC-mediated activation of alloreactive T cells. Methods and Results: We designed a CAR construct containing an scFv targeting OX40L, a CD28 costimulatory domain, along with CD3ζ, as well as the fluorescent reporter NeonGreen, all under the control of a synthetic FOXP3 promoter. This design enabled the selective and sustained expression of the anti-OX40L CAR on CD25+CD127- FoxP3+ Treg cells. We also created Tregs transduced with a non-targeting NeonGreen-only control construct (Neon-Tregs). Engineered OX40L CAR-Tregs maintained expression of the canonical Treg markers FOXP3, CD25, and CTLA-4, and demonstrated robust activation in the presence of OX40L-expressing cells. In co-culture assays of OX40L CAR-Tregs with OX40L-expressing K562 target cells, we demonstrate upregulation of Treg suppressive proteins CTLA-4, LAP, CD71, GARP and LAG3 without inducing expression of pro-inflammatory cytokines (no upregulation of IFNγ, TNFα, IL17a) (Figure 1A). Activation of Tregs via CAR recognition of OX40L+ cells resulted in 2.5-fold enhancement in their potency to suppress anti-CD3/CD28-driven T cell proliferation in vitro compared to Neon-Tregs at a 1:4 Treg-to-Teff ratio (p<0.05). Furthermore, OX40L-CAR-Tregs demonstrated a greater capacity to inhibit monocyte-derived dendritic cell activation, highlighting the superior suppressive activity of these OX40L-CAR-Tregs compared to non-targeting Neon-Tregs. Importantly, in a human xenograft model of GvHD, induced by transplantation of human PBMC into sublethally irradiated NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice, OX40L CAR-Treg demonstrated superior GvHD disease control compared to both PBMC alone and Neon-Tregs. Clinical scores were assessed by a combinatorial system including five parameters: weight loss, posture, mobility, skin, and fur conditions, and curves were compared using a 2-way ANOVA mixed-effects analysis, which demonstrated improvement in clinical xeno-GvHD with the OX40L CAR-Treg > Neon-Treg > PBMC alone over the entire length of analysis (p <0.001). An example of the substantial improvement in the clinical score on Day +14 is as follows: combined GvHD clinical score = 6.5 for PBMC vs 2.5 for Neon-Treg vs 0.15 for OX40L CAR-Treg, p <0.0001 for all comparisons. This control of clinical GvHD resulted in prolonged recipient survival when compared to both PBMC alone and to control Neon-Tregs (MST = 34 days for OX40L-CAR-Treg, 20 days for Neon-Treg, 14 days for PBMC alone, p<0.001, Figure 1B). Conclusions: Collectively, these results demonstrate a novel and efficacious approach using OX40L-CAR Tregs to enhance Treg suppressive function and to control GvHD. Because OX40L is upregulated on APCs in inflammatory environments, and its expression is limited to APCs and activated endothelial cells, this approach provides a unique strategy for the control of allo-immunity after HCT. Moreover, these OX40L-CAR Treg may be broadly applicable beyond HCT, to control allo-immunity after solid organ transplant and to suppress T cell activation in autoimmune diseases. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

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
文献互助1发布了新的文献求助10
刚刚
刚刚
天天快乐应助聪慧代芹采纳,获得10
刚刚
陈明宇完成签到,获得积分10
1秒前
YYY完成签到,获得积分10
1秒前
一多完成签到 ,获得积分20
1秒前
MHK完成签到,获得积分10
1秒前
舍得完成签到,获得积分10
1秒前
2秒前
Tammy完成签到 ,获得积分10
2秒前
Hello~完成签到,获得积分10
2秒前
2秒前
tent01完成签到,获得积分10
2秒前
科目三应助尹小末采纳,获得10
2秒前
3秒前
贪玩丸子完成签到,获得积分10
3秒前
1661321476完成签到,获得积分10
3秒前
Reef完成签到,获得积分10
3秒前
魏骜琦完成签到,获得积分10
4秒前
无花果应助iwww采纳,获得10
4秒前
张思琪完成签到,获得积分10
4秒前
4秒前
CipherSage应助无心的星月采纳,获得10
4秒前
Han完成签到,获得积分10
5秒前
猫好好完成签到,获得积分10
5秒前
廉不可发布了新的文献求助10
6秒前
Eureka完成签到 ,获得积分10
7秒前
8秒前
蔚蔚蓝天完成签到,获得积分10
8秒前
追寻依风完成签到,获得积分10
8秒前
8秒前
ddsyg126完成签到,获得积分10
8秒前
有魅力的从凝完成签到,获得积分10
8秒前
史呆芬发布了新的文献求助10
8秒前
欣慰的白羊完成签到,获得积分10
9秒前
9秒前
10秒前
煎饼煎饼完成签到,获得积分10
10秒前
淡然觅荷完成签到 ,获得积分10
10秒前
jack1511完成签到,获得积分10
10秒前
高分求助中
【提示信息,请勿应助】关于scihub 10000
A new approach to the extrapolation of accelerated life test data 1000
徐淮辽南地区新元古代叠层石及生物地层 500
Coking simulation aids on-stream time 450
北师大毕业论文 基于可调谐半导体激光吸收光谱技术泄漏气体检测系统的研究 390
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
Robot-supported joining of reinforcement textiles with one-sided sewing heads 360
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 4015970
求助须知:如何正确求助?哪些是违规求助? 3555964
关于积分的说明 11319479
捐赠科研通 3289040
什么是DOI,文献DOI怎么找? 1812373
邀请新用户注册赠送积分活动 887882
科研通“疑难数据库(出版商)”最低求助积分说明 812044