Journal: Nature Communications

Article Title: PTEN expression by an oncolytic herpesvirus directs T-cell mediated tumor clearance

doi: 10.1038/s41467-018-07344-1

Figure Lengend Snippet: HSV-P10 construction and characterization. a Graphical representation of PTEN and PTENα coding sequences showing the mutations engineered in PTENα start codons to enhance translation of full-length PTENα. b – c Graphical representation of the DNA structure of wild-type F-strain HSV1 backbone showing doubly deleted γ34.5 genes, and gene-disrupting insertional ICP6 locus containing an enhanced green fluorescent protein (eGFP) gene with PTENα insertion (HSV-P10) or without PTENα (HSVQ). d PTENα production in infected tumor cell lysates over time. The indicated tumor cells were infected with HSV-P10 at MOI = 0.5 and harvested 0–12 hpi and probed for PTEN by western blot

Article Snippet: Primary (from CST: PTEN (9559), Akt (4691), pAkt-S473 (9271), from abcam: HSV1 ICP4 (ab6514), GAPDH (ab9484)) and secondary antibodies (from abcam: anti-mouse HRP (ab6789), anti-rabbit HRP (ab6721) can be found in Supplementary Data .

Techniques: Infection, Western Blot

Journal: Nature Communications

Article Title: PTEN expression by an oncolytic herpesvirus directs T-cell mediated tumor clearance

doi: 10.1038/s41467-018-07344-1

Figure Lengend Snippet: HSV-P10 induces enhanced immune cell influx towards infected tumors. Tumor-bearing FVB/N mice were treated with 1e5 pfu of HSVQ, HSV-P10, or saline control 8 days post tumor implantation. a Stitched 4X bright field images of H&E stained sagittal sections of mouse brains 3 days post virus injection. b Representative 20X bright field images of H&E, Keratin 8, F4/80, NKp46, CD3, CD8α, PD-L1, and HSV1 (GFP) from sagittal sections of mouse brains 3 days post virus injection. Scale = 0.1 mm. c Ratio of macrophages (CD11b + F4/80 + CD45 bright ) to microglia (CD11b + F4/80 + CD45 dim ). d Percentage of cells in c positive for MHC-II. e Dendritic cell (CD11c + CD80 + MHC-II + ) infiltration. f NK cell (CD49b + NKp46 + ) infiltration. g CD8 + T-cell (CD3 + CD4 + ) infiltration. h CD4 + T-cell (CD3 + CD8 + ) infiltration. Data shown are averages ± s.d ( n = 3/group) Statistical significance was assessed by one-way ANAOVA ( n = 3, * p < 0.05, *** p < 0.001, **** p < 0.0001). Gating strategies for ( c – h ) are decribed in supplementary figure

Article Snippet: Primary (from CST: PTEN (9559), Akt (4691), pAkt-S473 (9271), from abcam: HSV1 ICP4 (ab6514), GAPDH (ab9484)) and secondary antibodies (from abcam: anti-mouse HRP (ab6789), anti-rabbit HRP (ab6721) can be found in Supplementary Data .

Techniques: Infection, Tumor Implantation, Staining, Injection

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: bioRxiv

Article Title: Th1 effector CD4 T cells rely on IFN-γ production to induce alopecia areata

doi: 10.1101/2025.05.07.650678

Figure Lengend Snippet: (A to C) In vitro activated CD8 T cells isolated from the SDLNs of AA mice were intradermally injected into recipient mice receiving 400μg intraperitoneal injections of isotype control or αCD4 antibodies (n=14 isotype control, n=15 αCD4). Mice were injected twice weekly for the duration of the experiment. (A) Schematic showing experimental design. (B) Representative images of ventral surface of mice in each group. (C) Kaplan-Meier disease curve for development of hair loss. ns=not significant, Log-rank test. (D to F) In vitro activated CD4 T cells isolated from the SDLNs of AA mice were intradermally injected into recipient mice receiving 400μg intraperitoneal injections of isotype control or αCD8β antibodies (n=10 each group). Mice were injected twice weekly for the duration of the experiment. (D) Schematic showing experimental design. (E) Representative images of ventral surface of mice in each group. (F) Kaplan-Meier disease curve showing development of hair loss. ***p=0.0003, Log-rank test. Data representative of two to three combined experiments.

Article Snippet: Anti-CD4 (BioXCell, BE0003), anti-CD8β (Lyt 3.2, BioXCell, BE0223), or isotype control IgG (BioXCell) were administered by intraperitoneal injection (400μg) two times weekly for the duration of the experiments.

Techniques: In Vitro, Isolation, Injection, Control