Journal: Neoplasia (New York, N.Y.)

Article Title: Screening and identification of HLA-A2-restricted neoepitopes for immunotherapy in endocrine therapy-resistant breast cancer.

doi: 10.1016/j.neo.2025.101200

Figure Lengend Snippet: Fig. 2. Immunoreactivity of mutant peptides targeting T2A2 cells in vitro. CTLs were induced with autologous MUT peptide-pulsed DCs from peripheral blood lymphocytes of HLA-A2+ healthy donors. A-B. CTLs were collected and co- cultured with T2A2 cells loaded with MUT/WT peptide and then were detected for IFN-γ production and FasL expression. C. CTLs were co-cultured with T2A2 cells that were pulsed with MUT peptide or corresponding WT peptide at an effector/target (E/T) ratio of 12.5:1, 25:1, and 50:1, respectively. LDH cytotoxicity killing assay was used to detect the immunogenicity of mutated epitope peptides. The T2A2 cells loaded with WT peptide group was defined as the negative control group. Data were represented as means ± SD. Statistical significance was determined by unpaired Student’s t-test, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Human breast cancer cell lines MDA-MB-231 (HLA-A*0201-positive) and MCF7 (HLA-A*0201-positive), along with the human lymphoblast cell line T2A2 (transporter associated with antigen processing deficient) were obtained from the American Type Culture Collection (ATCC, USA) and maintained under standard culture conditions in our laboratory.

Techniques: Mutagenesis, In Vitro, Cell Culture, Expressing, Immunopeptidomics, Negative Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Screening and identification of HLA-A2-restricted neoepitopes for immunotherapy in endocrine therapy-resistant breast cancer.

doi: 10.1016/j.neo.2025.101200

Figure Lengend Snippet: Fig. 4. CTLs obtained by MUT peptide pool induction in HLA-A2.1/Kb transgenic mice are able to specifically distinguish antigenic epitopes loaded on T2A2 cells. Immunogenicity of the MUT peptide in HLA-A2.1/Kb transgenic mice. Splenocytes and lymph node cells from HLA-A2.1/Kb transgenic mice immunized with either a peptide pool (Peptide pool group) or normal saline (Vehicle group) in combination with CpG ODN 1826 (30 µg per mouse) were restimulated in vitro with MUT peptides for 5 days. Then CTLs were co-cultured with different target cells to detect the CTL response in (A-B) intracellular cytokine assay and (C) cytotoxicity assay. The T2A2 cells loaded with WT peptide pool and T2A2 cells loaded with MUT peptide pool were severed as stimulator cells and target cells. The T2A2 cells loaded with WT peptide pool group was defined as the negative control group. Data were represented as means ± SD. Statistical significance was determined by unpaired Student’s t-test, **P < 0.01, ***P < 0.001.

Article Snippet: Human breast cancer cell lines MDA-MB-231 (HLA-A*0201-positive) and MCF7 (HLA-A*0201-positive), along with the human lymphoblast cell line T2A2 (transporter associated with antigen processing deficient) were obtained from the American Type Culture Collection (ATCC, USA) and maintained under standard culture conditions in our laboratory.

Techniques: Transgenic Assay, Immunopeptidomics, Saline, In Vitro, Cell Culture, Cytokine Assay, Cytotoxicity Assay, Negative Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Screening and identification of HLA-A2-restricted neoepitopes for immunotherapy in endocrine therapy-resistant breast cancer.

doi: 10.1016/j.neo.2025.101200

Figure Lengend Snippet: Fig. 5. CTLs obtained from MUT peptide pool induction in HLA-A2.1/Kb transgenic mice specifically recognize single mutant epitopes loaded on T2A2 cells. Splenocytes and lymph node cells from HLA-A2.1/Kb transgenic mice immunized with either a peptide pool (Peptide pool group) or normal saline (Vehicle group) in combination with CpG ODN 1826 (30 µg per mouse) were restimulated in vitro with MUT peptides for 5 days. Then CTLs were co-cultured with different target cells to detect the CTL response with cytotoxicity assay. The T2A2 cells loaded with single WT peptide and T2A2 cells loaded with single MUT peptide were severed as stimulator cells and target cells. The T2A2 cells loaded with single WT peptide group was defined as the negative control group. Data were represented as means ± SD. Statistical significance was determined by unpaired Student’s t-test, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Human breast cancer cell lines MDA-MB-231 (HLA-A*0201-positive) and MCF7 (HLA-A*0201-positive), along with the human lymphoblast cell line T2A2 (transporter associated with antigen processing deficient) were obtained from the American Type Culture Collection (ATCC, USA) and maintained under standard culture conditions in our laboratory.

Techniques: Transgenic Assay, Mutagenesis, Saline, In Vitro, Cell Culture, Cytotoxicity Assay, Negative Control