Pathogenic Autoimmunity in Atherosclerosis Evolves From Initially Protective Apolipoprotein B₁₀₀-Reactive CD4⁺ T-Regulatory Cells

Background: Throughout the inflammatory response that accompanies atherosclerosis, autoreactive CD4⁺ T-helper cells accumulate in the atherosclerotic plaque. Apolipoprotein B₁₀₀ (apoB), the core protein of low-density lipoprotein, is an autoantigen that drives the generation of pathogenic T-helper type 1 (T_H1) cells with proinflammatory cytokine secretion. Clinical data suggest the existence of apoB-specific CD4⁺ T cells with an atheroprotective, regulatory T cell (T_reg) phenotype in healthy individuals. Yet, the function of apoB-reactive T_regs and their relationship with pathogenic T_H1 cells remain unknown.

Methods: To interrogate the function of autoreactive CD4⁺ T cells in atherosclerosis, we used a novel tetramer of major histocompatibility complex II to track T cells reactive to the mouse self-peptide apo B_978-993 (apoB⁺) at the single-cell level.

Results: We found that apoB⁺ T cells build an oligoclonal population in lymph nodes of healthy mice that exhibit a T_reg-like transcriptome, although only 21% of all apoB⁺ T cells expressed the T_reg transcription factor FoxP3 (Forkhead Box P3) protein as detected by flow cytometry. In single-cell RNA sequencing, apoB⁺ T cells formed several clusters with mixed T_H signatures that suggested overlapping multilineage phenotypes with pro- and anti-inflammatory transcripts of T_H1, T helper cell type 2 (T_H2), and T helper cell type 17 (T_H17), and of follicular-helper T cells. ApoB⁺ T cells were increased in mice and humans with atherosclerosis and progressively converted into pathogenic T_H1/T_H17-like cells with proinflammatory properties and only a residual T_reg transcriptome. Plaque T cells that expanded during progression of atherosclerosis consistently showed a mixed T_H1/T_H17 phenotype in single-cell RNA sequencing. In addition, we observed a loss of FoxP3 in a fraction of apoB⁺ T_regs in lineage tracing of hyperlipidemic Apoe^-/- mice. In adoptive transfer experiments, converting apoB⁺ T_regs failed to protect from atherosclerosis.

Conclusions: Our results demonstrate an unexpected mixed phenotype of apoB-reactive autoimmune T cells in atherosclerosis and suggest an initially protective autoimmune response against apoB with a progressive derangement in clinical disease. These findings identify apoB autoreactive T_regs as a novel cellular target in atherosclerosis.