Journal: NPJ Vaccines

Article Title: Repurposing anti-viral subunit and mRNA vaccines T cell immunity for intratumoral immunotherapy against solid tumors

doi: 10.1038/s41541-025-01131-y

Figure Lengend Snippet: Experimental design: C57BL/6 mice were prime-boost immunized intramuscularly with the HPV vaccine or unimmunized prior to TC-1 tumor subcutaneous challenge one week after boost. When tumor reached a volume between 50 and 100 m 3 , mice were treated six times with saline, HPV-vax alone (1/25 th of the human dose) or L1 165-173 peptide (0.5 μg) admixed with polyI:C LMW (50 μg). Mice were monitored twice a week for ( A , C ) tumor growth and (B, D) survival. A , C Tumor growth is shown as the mean of tumor volume for each group and SE ( n = 6–9 per group). Statistical analysis with Dunn’s test was performed for multiple comparison of tumor growth. B , D Survival comparisons were assessed by Mantel–Cox test (**** P < 0.0001, *** P < 0.001, ** P < 0.01 * P < 0.05 n.s.: not significant). E IFN-γ production was assessed after IT treatment by intracellular staining of circulating CD8 + T cells after in vitro restimulation with L1 165-173 peptide. Data are shown as individual values and mean of cytokine production of CD44 + CD8 + T cells with SE. F Circulating anti-tumor E7-specific CD8 + T cell responses were measured by dextramer staining after IT treatment. Data are shown as individual percentage and mean of H2-D b /E7 49-57 + CD44 + CD8 + T cells. G anti-HPV16 L1 VLP antibody responses were measured by ELISA in plasma samples. Data are shown as individual and geometric mean of IgG EC50 titer and 95% confidence interval. H Cytokine and chemokine production was measured using a bead-based multiplex immunoassay in tumor lysates. Heatmap shows the average Z-score of the amount of each analyte per 50 μg of protein. I – L The cellular infiltrate was assessed by flow cytometry. Data are shown as individual values and mean percentage within CD45 + cells of ( I ) activated CD8 + T cells defined as CD8 + CD44 + CD62L - PD1 + , ( J ) myeloid cells defined as CD11b + , and percent within CD11b + cells of ( K ) neutrophils defined as CD11b + Ly6G + Ly6C int and ( L ) macrophages defined as CD11b + F480 + Ly6G - Ly6C int . Data are shown as individual values and mean percentage of cells within indicated gated population and SE. Statistical analysis (* P < 0.05 or numeric values) was performed using Dunn’s test for multiple comparison between groups ( n = 4–5 per group).

Article Snippet: Mice were injected with 200 μg of an anti-mouse CD8 antibody (clone 2.43, BioXcell InVivoPlus) or a rat IgG2b control antibody (clone LTF-2, BioXcell InVivoPlus) diluted in 200 μl phosphate-buffered saline (PBS) on day 3 and 1 prior to the first IT injection and on day 1 prior to each subsequent IT injection.

Techniques: Saline, Comparison, Staining, In Vitro, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Multiplex Assay, Flow Cytometry

Journal: NPJ Vaccines

Article Title: Repurposing anti-viral subunit and mRNA vaccines T cell immunity for intratumoral immunotherapy against solid tumors

doi: 10.1038/s41541-025-01131-y

Figure Lengend Snippet: Experimental design: C57BL/6 mice were prime-boost immunized intramuscularly with the VZV vaccine prior to TC-1 tumor subcutaneous challenge one week after boost. When tumor reached a volume between 50 and 100 m , mice were treated six times with saline, VZV-vax alone (1/40 th of the human dose) or the MHC-II-restricted gE 71-90 peptide (2 μg) admixed with polyI:C LMW (50 μg). Mice were monitored twice a week for ( A ) tumor growth and ( B ) survival. A Tumor growth is shown as the mean of tumor volume for each group and SE ( n = 7 per group). Statistical analysis with Dunn’s test was performed for multiple comparison of tumor growth. B Survival comparisons were assessed by Mantel–Cox test (** P < 0.01, * P < 0.05). C Tumor-free mice from the VZV-vax and gE 71-90 + polyI:C-treated groups, and naïve mice were rechallenged with TC-1 cells, 60 days after the primary challenge and tumor growth was monitored twice a week until endpoint. D anti-VZV gE protein responses were measured by ELISA in plasma samples. Data are shown as individual values and geometric mean of the EC50 of IgG titer and 95% confidence interval. In a separate experiment, ( E ) IFN-γ, ( F ) TNF-α and ( G ) IL-2 production was assessed 36 h after the last IT treatment by intracellular staining of circulating CD4 + T cells after in vitro restimulation with VZV gE overlapping peptide library. Individual cytokine production is shown as individual values and mean percentage within CD44 + CD4 + T cells with SE. Statistical analysis was performed using Dunn’s test for multiple comparison between groups (* P < 0.05 ** P < 0.01, *** P < 0.001, **** P < 0.0001, numeric values, ns = not significant) ( n = 4 per group).

Article Snippet: Mice were injected with 200 μg of an anti-mouse CD8 antibody (clone 2.43, BioXcell InVivoPlus) or a rat IgG2b control antibody (clone LTF-2, BioXcell InVivoPlus) diluted in 200 μl phosphate-buffered saline (PBS) on day 3 and 1 prior to the first IT injection and on day 1 prior to each subsequent IT injection.

Techniques: Saline, Comparison, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Staining, In Vitro

Journal: The EMBO Journal

Article Title: A bacterial network of T3SS effectors counteracts host pro-inflammatory responses and cell death to promote infection

doi: 10.1038/s44318-025-00412-5

Figure Lengend Snippet: ( A , B ) Unprimed primary BMDMs, ( C , D ) immortalised BMDMs or ( E – H ) unprimed primary BMDMs were infected with log-phase C. rodentium ( Cr ) and/or Δ nleE for ( A , E ) 5 h or ( B – D , F – H ) the indicated time points. ( G , H ) BMDMs were treated with 100 μg/ml α-TNF neutralising antibody or IgG isotype control for 30 min before Cr infection. ( I – K ) CMT-93 cells were unprimed or primed with LPS (100 ng/ml) or TNF (100 ng/ml) for 3 h and infected with Cr and Δ nleE over time ( I , K ) or for ( J ) 5 h. ( A , C , E ) LDH release was quantified. ( B , D , F , J ) Mixed supernatant and lysates were examined by immunoblotting. ( G ) SYTOX Green uptake and ( H , I , K ) caspase-3/7 activity (DEVD-positive) were quantified using IncuCyte. Where indicated, cells were treated with Nec-1s (100 μM) for 30 min before infection. ( A ) Pooled data are mean ± SEM of four independent experiments ( P ≤ 0.0001 for both conditions). ( C ) Pooled data are mean ± SEM of three independent experiments ( P = 0.0010). ( E ) Pooled data are mean ± SEM of three independent experiments ( P = 0.0257 for Tnfr1 −/− and P = 0.0094 for Trif Lps2 ). Data are mean ± SD of technical ( G , H ) duplicates and ( I , K ) triplicates representative of three independent experiments. All P values were calculated with two-way ANOVA test. Data are considered significant when P ≤ 0.05, with * P ≤ 0.05, ** P ≤ 0.01 or **** P ≤ 0.0001. .

Article Snippet: InVivoMab rat IgG1 isotype control (anti-HRP) , Bio X Cell , #BE0088; RRID:AB_1107775.

Techniques: Infection, Control, Western Blot, Activity Assay

Journal: The EMBO Journal

Article Title: A bacterial network of T3SS effectors counteracts host pro-inflammatory responses and cell death to promote infection

doi: 10.1038/s44318-025-00412-5

Figure Lengend Snippet: Reagents and tools table

Article Snippet: InVivoMab rat IgG1 isotype control (anti-HRP) , Bio X Cell , #BE0088; RRID:AB_1107775.

Techniques: Recombinant, Control, Sequencing, Cloning, Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Software, Imaging, Cell Analysis