Fig. 3

Clinical and immune responses to personalized Neo-DCVac in patient 1 with metastatic lung adenocarcinoma. a Clinical course of the disease and previous treatments of patient 1. The patient failed three lines of chemotherapy and radiotherapy but remained without any signs of disease progression for 7.6 months following Neo-DCVac administration. b Computed tomography (CT) scans were performed before and after five doses of personalized Neo-DCVac immunotherapy, and representative images are shown. c Autologous PBMCs were stimulated with 13 candidate mutant peptides for 10 days, after which IFN-γ ELISpot assays were performed to assess the T-cell-specific antigen response. One percent phytohemagglutinin (PHA) and no peptide stimulation represent the positive and negative controls, respectively. The IFN-γ ELISpot picture of fold changes of mutant peptides/WT peptides >2 is shown. d Before and after Neo-DCVac treatment, the PBMCs were stimulated with neoantigen peptides overnight, and the concentrations of IL-2, IFN-γ, and TNF-α in the culture supernatants were measured by cytometric bead array (CBA). Data are representative of results from three independent experiments (n = 3). This data are shown as the mean ± s.e.m. e After Neo-DCVac treatment, the concentrations of IL-2, IFN-γ, and TNF-α in the culture supernatant were measured by CBA. The data represent the results of three independent experiments. f Analysis of ex vivo T-cell responses to neoantigen peptides after overnight exposure to neoepitopes and corresponding WT peptides using intracellular cytokine staining followed by flow cytometry. g Ex vivo intracellular cytokine staining of PBMCs after neoepitope stimulation. PBMCs were pre-gated on CD3⁺ and CD4⁺ T cells. Cytokine-producing neoantigen-reactive cells express CD45RO and PD-1, demonstrating an antigen-experienced memory T-cell phenotype (****P < 0.0001)