Latency-associated peptide (LAP)+CD4+ regulatory T cells prevent atherosclerosis by modulating macrophage polarization

Jian Yu; Wenbin Xu; Qian Dong; Qingwei Ji; Min Cheng; Desheng Hu; Yifan Cai; Qiutang Zeng; Kunwu Yu

doi:10.1016/j.clim.2023.109767

Latency-associated peptide (LAP)⁺CD4⁺ regulatory T cells prevent atherosclerosis by modulating macrophage polarization

Clin Immunol. 2023 Oct:255:109767. doi: 10.1016/j.clim.2023.109767. Epub 2023 Sep 7.

Authors

Jian Yu¹, Wenbin Xu¹, Qian Dong¹, Qingwei Ji², Min Cheng¹, Desheng Hu³, Yifan Cai¹, Qiutang Zeng⁴, Kunwu Yu⁵

Affiliations

¹ Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; Hubei Key Laboratory of Biological Targeted Therapy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, China; Hubei Provincial Engineering Research Center of Immunological Diagnosis and Therapy for Cardiovascular Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, China.
² Department of Cardiology, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, China.
³ Department of Integrated Traditional Chinese and Western Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
⁴ Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; Hubei Key Laboratory of Biological Targeted Therapy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, China; Hubei Provincial Engineering Research Center of Immunological Diagnosis and Therapy for Cardiovascular Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, China. Electronic address: zengqt139@sina.com.
⁵ Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; Hubei Key Laboratory of Biological Targeted Therapy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, China; Hubei Provincial Engineering Research Center of Immunological Diagnosis and Therapy for Cardiovascular Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, China. Electronic address: bushyukunwu@126.com.

PMID: 37689092
DOI: 10.1016/j.clim.2023.109767

Abstract

Rationale: A persistent autoimmune and inflammatory response plays a critical role in the progression of atherosclerosis. The transcription factor forkhead box P3 (Foxp3)⁺CD4⁺ regulatory T cells (Foxp3⁺ Tregs) attenuate atherosclerosis. Latency-associated peptide (LAP)⁺CD4⁺ T cells are a new class of Tregs whose role in atherosclerosis is unknown.

Objective: To investigate the function of CD4⁺LAP⁺ Tregs in inhibiting inflammation and preventing atherosclerosis.

Methods and results: Depletion of CD4⁺LAP⁺ Tregs results in aggravated inflammation and atherosclerotic lesions. Mechanistically, CD4⁺LAP⁺ Treg depletion was associated with decreased M2-like macrophages and increased Th1 and Th17 cells, characterized by increased unstable plaque promotion and decreased expression of inflammation-resolving factors in both arteries and immune organs. In contrast, adoptive transfer of CD4⁺LAP⁺ Tregs to ApoE^-/- mice or CD4^-/-ApoE^-/- mice led to decreased atherosclerotic lesions. Compared with control animals, adoptive transfer of CD4⁺LAP⁺ Tregs induced M2-like macrophage differentiation within the atherosclerotic lesion and spleen, associated with increased collagen and α-SMA in plaques and decreased expression of MMP-2 and MMP-9. Mechanistic studies reveal that isolated CD4⁺LAP⁺ Tregs exhibit a tolerance phenotype, with increased expression of inhibitory cytokines and coinhibitory molecules. After coculture with CD4⁺LAP⁺ Tregs, monocytes/macrophages display typical features of M2 macrophages, including upregulated expression of CD206 and Arg-1 and decreased production of MCP-1, IL-6, IL-1β and TNF-α, which was almost abrogated by transwell and partially TGF-β1 neutralization. RNA-seq analysis showed different gene expression profiles between CD4⁺LAP⁺ Tregs and LAP^-CD4⁺ T cells and between CD4⁺LAP⁺ Tregs of ApoE^-/- mice and CD4⁺LAP⁺ Tregs of C57BL/6 mice, of which Fancd2 and IL4i1 may contribute to the powerful inhibitory properties of CD4⁺LAP⁺ Tregs. Furthermore, the number and the suppressive properties of CD4⁺LAP⁺ Tregs were impaired by oxLDL.

Conclusions: Our data indicate that the remaining CD4⁺LAP⁺ Tregs play a protective role in atherosclerosis by modulating monocyte/macrophage differentiation and regulatory factors, which may partly explain the protective effect of T cells tolerance in atherosclerosis. Moreover, adoptive transfer of CD4⁺LAP⁺ Tregs constitutes a novel approach to treat atherosclerosis.

Keywords: Atherosclerosis; Latency-associated peptide; M2-type macrophage; Regulatory T cells.