Background: CD (cluster of differentiation) 4 ⁺ T-cell responses to APOB (apolipoprotein B) are well characterized in atherosclerotic mice and detectable in humans. CD4 ⁺ T cells recognize antigenic peptides displayed on highly polymorphic HLA (human leukocyte antigen)-II. Immunogenicity of individual APOB peptides is largely unknown in humans. Only 1 HLA-II-restricted epitope was validated using the DRB1*07:01-APOB _{3036 –3050} tetramer. We hypothesized that human APOB may contain discrete immunodominant CD4 ⁺ T-cell epitopes that trigger atherosclerosis-related autoimmune responses in donors with diverse HLA alleles. Methods: We selected 20 APOB-derived peptides (APOB ₂₀ ) from an in silico screen and experimentally validated binding to the most commonly occurring human HLA-II alleles. We optimized a restimulation-based workflow to evaluate antigenicity of multiple candidate peptides in HLA-typed donors. This included activation-induced marker assay, intracellular cytokine staining, IFNγ (interferon gamma) enzyme–linked immunospot and cytometric bead array. High-throughput sequencing revealed TCR (T-cell receptor) clonalities of APOB-reactive CD4 ⁺ T cells. Results: Using stringent positive, negative, and crossover stimulation controls, we confirmed specificity of expansion-based protocols to detect CD4 ⁺ T cytokine responses to the APOB ₂₀ pool. Ex vivo assessment of AIM ⁺ CD4 ⁺ T cells revealed a statistically significant autoimmune response to APOB ₂₀ but not to a ubiquitously expressed negative control protein, actin. Resolution of CD4 ⁺ T responses to the level of individual peptides using IFNγ enzyme–linked immunospot led to the discovery of 6 immunodominant epitopes (APOB ₆ ) that triggered robust CD4 ⁺ T activation in most donors. APOB ₆ -specific responding CD4 ⁺ T cells were enriched in unique expanded TCR clonotypes and preferentially expressed memory markers. Cytometric bead array analysis detected APOB ₆ -induced secretion of both proinflammatory and regulatory cytokines. In clinical samples from patients with angiographically verified coronary artery disease, APOB ₆ stimulation induced higher activation and memory phenotypes and augmented secretion of proinflammatory cytokines TNF (tumor necrosis factor) and IFNγ, compared with patients with low coronary artery disease. Conclusions: Using 3 cohorts, each with ≈20 donors, we discovered and validated 6 immunodominant, HLA-II–restricted APOB epitopes. The immune response to these APOB epitopes correlated with coronary artery disease severity.