human ifn γ elispot plus kit  (Mabtech Inc)

Journal: Frontiers in Immunology

Article Title: Neoantigen peptide-pulsed dendritic cell vaccine therapy after surgical treatment of pancreatic cancer: a retrospective study

doi: 10.3389/fimmu.2025.1571182

Figure Lengend Snippet: Protocol for intranodal Neo-P DC vaccine therapy. Predicted neoantigens based on single-nucleotide variants identified by NGS were synthesized, and DCs differentiated from patient-derived PBMCs were pulsed by synthesized neoantigen peptides to generate Neo-P DC vaccines. The Neo-P DC vaccines were directly administered into the lymph nodes under US guidance. After the administration of three vaccines at 2-week intervals, IFN-γ ELISpot analysis was performed, and Neo-P+ DCs were used for the following three vaccines. After administering the six DC vaccines, ELISpot responses were measured again. PBMCs and plasma were cryopreserved before, during, and after treatment. NGS, next-generation sequencing; DCs, dendritic cells; PBMCs, peripheral blood mononuclear cells; US, ultrasound; IFN, interferon; ELISpot, enzyme-linked immunospot; Neo-P DC, neoantigen peptide-pulsed dendritic cell; NeoP+ DC, IFN-γ ELISpot-positive neoantigen peptide-pulsed dendritic cell.

Article Snippet: Human IFN-γ ELISpot Plus Kit (Mabtech Inc., Cincinnati, OH, USA) was used as previously described ( ).

Techniques: Immunopeptidomics, Synthesized, Derivative Assay, Vaccines, Enzyme-linked Immunospot, Clinical Proteomics, Next-Generation Sequencing

Journal: Frontiers in Immunology

Article Title: Neoantigen peptide-pulsed dendritic cell vaccine therapy after surgical treatment of pancreatic cancer: a retrospective study

doi: 10.3389/fimmu.2025.1571182

Figure Lengend Snippet: Profiles of Neoantigens and Immunological Responses (Post-operative recurrence cases).

Article Snippet: Human IFN-γ ELISpot Plus Kit (Mabtech Inc., Cincinnati, OH, USA) was used as previously described ( ).

Techniques: Immunopeptidomics, Enzyme-linked Immunospot

Journal: Frontiers in Immunology

Article Title: Neoantigen peptide-pulsed dendritic cell vaccine therapy after surgical treatment of pancreatic cancer: a retrospective study

doi: 10.3389/fimmu.2025.1571182

Figure Lengend Snippet: Profiles of Neoantigens and Immunological Responses (Adjuvant cases).

Article Snippet: Human IFN-γ ELISpot Plus Kit (Mabtech Inc., Cincinnati, OH, USA) was used as previously described ( ).

Techniques: Immunopeptidomics, Adjuvant, Enzyme-linked Immunospot

Journal: Frontiers in Immunology

Article Title: Neoantigen peptide-pulsed dendritic cell vaccine therapy after surgical treatment of pancreatic cancer: a retrospective study

doi: 10.3389/fimmu.2025.1571182

Figure Lengend Snippet: Neoantigen-pulsed DC vaccine reliably induced neoantigen-specific T cells. Immune responses of peripheral blood lymphocytes to neoantigen peptides before and after Neo-P DC vaccine therapy in (A) Recurrence Case #7 and (B) Recurrence Prevention Case #1. Each panel shows the IFN-γ ELISpot response to neoantigen peptide measured in peripheral blood lymphocytes from each patient [Ly, lymphocytes alone; Ly + DC, lymphocytes + dendritic cells; Ly + DC + pep (I), lymphocytes + dendritic cells + neoantigen peptide I (class I peptide); Ly + DC + pep (II), lymphocytes + dendritic cells + neoantigen peptide II (class II peptide). (C) (Upper) Results of ELISpot responses to class I peptides in all 16 cases. (Middle) Percentages of responders and ELISpot-positive peptides among all 109 class I peptides. (Lower) Percentages in each case of recurrence and adjuvant therapy. (D) (Upper) Results of ELISpot responses to class II peptides in all five cases. (Middle) Percentages of responders and ELISpot-positive peptides among all 16 class II peptides. (Lower) Percentages in each case of recurrence and adjuvant therapy. ELISpot, enzyme-linked immunospot.

Article Snippet: Human IFN-γ ELISpot Plus Kit (Mabtech Inc., Cincinnati, OH, USA) was used as previously described ( ).

Techniques: Enzyme-linked Immunospot, Adjuvant

Journal: Frontiers in Immunology

Article Title: Neoantigen peptide-pulsed dendritic cell vaccine therapy after surgical treatment of pancreatic cancer: a retrospective study

doi: 10.3389/fimmu.2025.1571182

Figure Lengend Snippet: Recurrence Case #8 treated with long-peptide-combined Neo-P DC vaccine shows the importance of neoantigen-specific CD4 + T cells. (A) Recurrence Case #8. (B) Immune responses of peripheral blood lymphocytes to both short neoantigen peptides (class I peptides) and long neoantigen peptides (class II peptides) in Recurrence Case #8. The panel shows the IFN-γ ELISpot response to neoantigen peptides measured in peripheral blood lymphocytes [Ly, lymphocytes alone; Ly + DC, lymphocytes + dendritic cells; Ly + DC + pep (I) , lymphocytes + dendritic cells + peptide]. (C) Graph of IFN-γ spot number in (B) . Note the marked responses of T cells to class II affinity long neoantigen (pep 6). (D) Computed tomography image showing the change in metastatic lesion before and after Neo-P DC vaccine therapy. (E) Analysis of TCRβ repertoire for pre- and post-vaccine T cells in peripheral blood. The top six most frequent clones are listed in pre-Vaccine, and the top seven most frequent clones are listed in post-Vaccine. DI, inverse Simpson’s diversity index. (F) Analysis of TCRβ repertoire for CD8 + T cells sorted from post-vaccine peripheral blood mononuclear cell–derived lymphocytes expanded by being pulsed with pep 6. The top six most frequent clones are listed. (G) Frequency of the top six enriched CD8 + T cell clones (F) in pre- and post-vaccine peripheral blood. (H) Analysis of TCRβ repertoire for CD4 + T cells sorted from post-vaccine peripheral blood mononuclear cell–derived lymphocytes expanded by being pulsed with pep 6. The top five most frequent clones are listed. (I) Frequency of the top five enriched CD4 + T cell clones (H) in pre- and post-vaccine peripheral blood. (J) Immunohistochemical analysis of primary tumor (Recurrence Case #8) for HE, CD20 (B-cell marker), and HLA-DR. Scale bars, 100 µm. IFN-γ ELISpot, interferon-γ–based enzyme-linked immunospot; Neo-P DC, neoantigen peptide-pulsed dendritic cell; TCR, T-cell receptor. * in (E, H, I) refers to the most expanded clone by pulsed with pep6.

Article Snippet: Human IFN-γ ELISpot Plus Kit (Mabtech Inc., Cincinnati, OH, USA) was used as previously described ( ).

Techniques: Enzyme-linked Immunospot, Computed Tomography, Clone Assay, Derivative Assay, Immunohistochemical staining, Marker