# Project 4: APOB-specific CD4 and CD8 T cells exacerbate atherosclerosis

> **NIH NIH P01** · LA JOLLA INSTITUTE FOR IMMUNOLOGY · 2022 · $43,008

## Abstract

Project 4 Summary
Project 4 in this PPG with 4 projects and 3 cores is focused on the T cell response to apolipoprotein B (ApoB),
an atherosclerosis antigen, in humans and mice. The central hypothesis is that the autoimmune response
is initially atheroprotective, but becomes pro-atherogenic (switches) as atherosclerosis progresses.
We will address this in humans with and without coronary artery disease (Cardiovascular Assessment Virginia
[CAVA] and CT-CAVA cohorts) and in newly generated Apoe-/- FoxP3-ERT2-Cre-GFP ROSA26-fl-STOP-fl-
RFP lineage tracker mice. We propose 3 specific aims. Aim 1 is to define the ApoB-specific CD4 and CD8 T
cells in PBMCs from CAD cases and controls. This includes T cell transcriptomes, TCRα and β sequences, cell
surface phenotypes by Ab-Seq, and finding the immunodominant ApoB epitopes to which they respond. The
main methods are single cell (sc) RNA-Seq, antibody (Ab)-Seq and T cell receptor (TCR)-Seq. To find the
immunodominant epitopes, of key relevance for cell-based and immunomodulatory therapeutics, we will use a
megapool of 208 MHC-binding human ApoB peptides, followed by systematic deconvolution tested by Elispot,
intracellular cytokine staining (ICS), cytokine capture assays (CCA) and antigen-induced marker (AIM) assays.
Aim 2 is to test how ApoB-specific T cells develop by combining the new mouse model with ApoB-specific
tetramers. Our preliminary data support the hypothesis that some ApoB-specific CD4 T cells already exist in
very young mice. Some of these cells are effector T cells (non-Tregs). Other FoxP3+ CD4+ Tregs in mice
switch their transcriptomes from atheroprotective to pro-atherogenic with progression of atherosclerosis.
Mechanistically, we will test the metabolic and epigenetic hypotheses of Treg to exTreg switch. Aim 3 is to test
the hypothesis that ApoB-specific T cells in humans with cardiovascular disease include Tregs, exTregs and
non-Tregs, using scRNA-Seq and deconvolution (CIBERSORT) methods. When the proposed work is done,
we will know the dominant MHC-II-restricted epitopes in human ApoB and the polarization of responding T
cells by scRNA-Seq, Ab-Seq and TCR-Seq. We will know the mechanism of Treg phenotype switching and the
proportion of Tregs, exTregs and other ApoB-specific T cells in PBMCs (CAVA and CT-CAVA cohorts) and
endarteriectomy transcriptomes (BIKE cohort).

## Key facts

- **NIH application ID:** 10334097
- **Project number:** 2P01HL136275-06
- **Recipient organization:** LA JOLLA INSTITUTE FOR IMMUNOLOGY
- **Principal Investigator:** Klaus F. Ley
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $43,008
- **Award type:** 2
- **Project period:** 2017-08-01 → 2022-06-02

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10334097

## Citation

> US National Institutes of Health, RePORTER application 10334097, Project 4: APOB-specific CD4 and CD8 T cells exacerbate atherosclerosis (2P01HL136275-06). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10334097. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*