Journal: bioRxiv

Article Title: A Neural Arming Niche in Tumor-Draining Lymph Nodes Programs CD8⁺T Cell Cytotoxicity via GZMB Norepinephrinylation

doi: 10.64898/2026.02.25.707881

Figure Lengend Snippet: a-e , Targeted liquid chromatography–tandem mass spectrometry (LC-MS/MS) analysis of a TGM2-transamidated monodansylcadaverine (MDC) to GZMB peptide 37-48 ( a ), peptide 77-89 ( b and c ), peptide 159-170 ( d ), peptide 177-187 ( e ). f . Representative FACS dot plots showing the apoptosis of KPC1199 tumor cells with/without co-culturing with EL4 cells. g , Mass spectrometry analysis was conducted on E3 ligase proteins that interacted with GZMB, the samples purified using GZMB antibody in Co-IP experiments.

Article Snippet: EL4 cells were purchased from ATCC.

Techniques: Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Purification, Co-Immunoprecipitation Assay

Journal: bioRxiv

Article Title: A Neural Arming Niche in Tumor-Draining Lymph Nodes Programs CD8⁺T Cell Cytotoxicity via GZMB Norepinephrinylation

doi: 10.64898/2026.02.25.707881

Figure Lengend Snippet: a, Targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of a TGM2-transamidated NE to GZMB peptide 37–48. b, EL4 cells were transfected with Flag-fused lentiviruses expressing either WT-GZMB or Gln43 to Ala (Q43A) mutated GZMB (Q43A-GZMB). Cells were stimulated with IL-2 and αCD3/αCD28 for 72 hours, Flag-beads were used to pull down Flag-GZMB, and the ubiquitination level of GZMB protein were detected in EL4 cells with overexpression of WT-GZMB or Q43A-GZMB (3 independent experiments). c, EL4 cells were stimulated with IL-2 and αCD3/αCD28 for 72 hours. After 72 hours, EL4 cells were co-cultured with KPC1199 tumor cells for 24 hours, and then flow cytometry was used to assess the levels of apoptosis in KPC1199 tumor cells. As a control, tumor cells incubation with wide-type EL4 cells were subjected to FACS analysis. Bar graph showed the quantification of apoptosis (EL4 cells: tumor cells = 5:1; n=3 per group; 3 independent experiments). d, EL4 cells were stimulated with IL-2 and αCD3/αCD28 for 72 hours. After 72 hours, cells were treated with inhibitor CHX at 0, 1, 2, 3 and 4 hours, and WT-GZMB or Q43A-GZMB protein levels were detected by western blot respectively. Left panels: Representative western blot images, β-actin was set as control. Right panels: Graph showed the relative intensity (3 independent experiments). e, Upper panels: Tgm2 fl/fl or Tgm2 fl/fl CD8-Cre CD8 + T cells were stimulated with IL-2 and αCD3/αCD28 for 72 hours, GZMB was immunoprecipitated and UHRF1 protein binding to GZMB was detected by western blot. GZMB was set as control. Lower panels: UHRF1 protein levels in Tgm2 fl/fl or Tgm2 fl/fl CD8-Cre CD8 + T cells were detected by western blot. β-actin was set as control, IP, immunoprecipitation (3 independent experiments). f, Upper panels: EL4 cells were stimulated with IL-2 and αCD3/αCD28 for 72 hours, Flag-GZMB was immunoprecipitated and UHRF1 protein binding to GZMB were detected by western blot. GZMB was set as control. Lower panels: UHRF1 protein levels in WT-GZMB (EL4) or Q43A-GZMB (EL4) cells were detected by western blot. β-actin was set as control, IP, immunoprecipitation (3 independent experiments). g, A ribbon trace of GZMB with labeled surface loops. The locations are shown for the catalytic residues, H64, D108, S203, and R228 (in yellow), and site of norepinephrinylation amino acids Q43 (in red). This model was from Uniprot (AF-P04187-F1-V4). h, TGM2 monoaminylation assay with GZMB protein in the absence (Vehicle) or presence of 5 μM NE. Activity of norepinephrinylated GZMB was analyzed. The immediately fluorescenceintensity at Ex/Em = 380/500 nm was measured in a microplate reader in kinetic mode for 30 min at 37°C protected from light (n = 3 per group, 3 independent experiments). Statistical significance was determined by two-tailed unpaired Welcht’s t -test (c, h), Error bars: SEM. ** P < 0.01; *** P < 0.001; N.S. , not significant.

Article Snippet: EL4 cells were purchased from ATCC.

Techniques: Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Transfection, Expressing, Ubiquitin Proteomics, Over Expression, Cell Culture, Flow Cytometry, Control, Incubation, Western Blot, Immunoprecipitation, Protein Binding, Labeling, Activity Assay, Two Tailed Test

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: synDNA vaccine against TCR chains and neoantigens for T cell lymphoma therapy

doi: 10.1007/s00262-026-04302-5

Figure Lengend Snippet: TCRfullvax design and characterization. ( A ) TCRfullvax design representing order in which each TCR chain was included. ( B ) Schematic representation of immunization schedule for TCRfullvax immunogenicity characterization. ( C ) Mean IFNγ spots generated against TCRα, TCRβ and TCRγ chains by the TCRfullvax. ( D ) IFNγ spots generated against each peptide pool derived from the TCRα chain. ( E ) IFNγ spots generated against each peptide pool derived from the TCRβ chain. ( F ) IFNγ spots generated against each peptide pool derived from the TCRγ chain. ( G ) Schematic of tumor challenge testing efficacy of TCRfullvax with EL4 cells in vivo. ( H ) Mean EL4 tumor sizes in mice treated with TCRfullvax or empty vector control pVax. ( I ) Survival of EL4 tumor bearing mice demonstrating statistically significant improvement in survival of mice treated with TCRfullvax

Article Snippet: The EL4 cell line was purchased from ATCC.

Techniques: Immunopeptidomics, Generated, Derivative Assay, In Vivo, Plasmid Preparation, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: synDNA vaccine against TCR chains and neoantigens for T cell lymphoma therapy

doi: 10.1007/s00262-026-04302-5

Figure Lengend Snippet: TCRfullvax-treated tumors have greater CD8+ T cell infiltration. ( A ) CD8+ T cell infiltration in tumors of mice immunized with either pVax or TCRfullvax. ( B ) %PD1 expression on CD8+ T cells in tumors of mice immunized with either pVax or TCRfullvax. ( C ) %KLRG1 expression on CD8+ T cells in tumors of mice immunized with either pVax or TCRfullvax. ( D ) PD1 MFI on CD8+ T cells in tumors of mice immunized with either pVax or TCRfullvax. ( E ) KLRG1 MFI on CD8+ T cells in tumors of mice immunized with either pVax or TCRfullvax. ( F ) %TCRβ+ TCRVβ12+ (TCR expressing EL4 cells) in tumors of mice immunized with either pVax or TCRfullvax. ( G ) Flow cytometry plots showing TCRβ+ TCRVβ12+ double positive cells in tumors of mice immunized with either pVax or TCRfullvax

Article Snippet: The EL4 cell line was purchased from ATCC.

Techniques: Expressing, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: synDNA vaccine against TCR chains and neoantigens for T cell lymphoma therapy

doi: 10.1007/s00262-026-04302-5

Figure Lengend Snippet: EL4neovax controls EL4 tumors in mice. ( A ) Schematic of tumor challenge testing efficacy of EL4neovax with EL4 cells in vivo. ( B ) Mean EL4 tumor sizes in mice treated with EL4neovax or empty vector control pVax. ( C ) Tumor size of each individual mouse treated with either EL4neovax or pVax. ( D ) Survival of EL4 tumor bearing mice demonstrating improvement in survival of mice treated with EL4neovax

Article Snippet: The EL4 cell line was purchased from ATCC.

Techniques: In Vivo, Plasmid Preparation, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: synDNA vaccine against TCR chains and neoantigens for T cell lymphoma therapy

doi: 10.1007/s00262-026-04302-5

Figure Lengend Snippet: Synergistic effect of TCRfullvax and EL4neovax. ( A ) Schematic of tumor challenge testing synergy of TCRfullvax and EL4neovax for controlling EL4 cells in vivo. ( B ) Mean EL4 tumor sizes in mice treated with TCRfullvax, EL4neovax, both vaccines co-delivered or empty vector control pVax. ( C ) Tumor size of each individual mouse treated with either TCRfullvax, EL4neovax, both vaccines co-delivered or pVax. ( D ) Survival of EL4 tumor bearing mice demonstrating improved survival of mice treated with combination therapy compared to either vaccine alone

Article Snippet: The EL4 cell line was purchased from ATCC.

Techniques: In Vivo, Vaccines, Plasmid Preparation, Control