rabbit polyclonal antibody targeting mhc class ii  (Bioss)

Journal: Molecular Therapy Oncology

Article Title: Oncolytic virotherapy mobilizes tumor-resident, granzyme B-producing bystander CD4 + T cells to inhibit systemic microbial infection

doi: 10.1016/j.omton.2026.201187

Figure Lengend Snippet: OV-BYTE strategy boosts systemic pathogen-specific CD4 + T MEM cell responses (A) Schematic of the experimental design. Congenic CD45.1 + SM CD4 + T cells were adoptively transferred into naive C57BL/6 recipients (CD45.2 + ), which were then infected with LCMV Armstrong and engrafted with MC38 cells on day 60 post infection. On days 7–12 after tumor engraftment, recipients were administered PBS, NDV-WT, or NDV-GP daily via the intratumoral route. On day 15 post-tumor engraftment, SM CD4 + T cells in various organs were analyzed. (B and C) Frequency (B) and number (C) of SM CD4 + cells from the indicated organs of groups administrated PBS (indicated by gray dots), NDV-WT (indicated by blue dots), or NDV-GP (indicated by red dots). Quantification of SM CD4 + cells from 100 μL of peripheral blood. (D) Flow cytometry analysis of SM CD4 + T cells on day 15 post-tumor engraftment from the PBS-, NDV-WT-, or NDV-GP-treated group. Numbers adjacent to the outlined areas indicates percentages of SM CD4 + cells among total CD4 + T cells. (E) Schematic of the experimental design. Naive C57BL/6 mice were infected with LCMV Armstrong and engrafted with MC38 cells on day 60 post infection. On days 7–12 after tumor engraftment, recipients were administered PBS, NDV-WT, or NDV-GP daily via the intratumoral route. On day 15 post-tumor engraftment, endogenous LCMV GP 66-77 tetramer + CD4 + T cells in various organs were analyzed. (F and G) Frequency (F) and number (G) of endogenous LCMV GP 66-77 tetramer + CD4 + T cells from the indicated organs of groups administrated PBS (indicated by gray dots), NDV-WT (indicated by blue dots), or NDV-GP (indicated by red dots). (H) Schematic of the experimental design. Congenic CD45.1 + OT-II CD4 + T cells were adoptively transferred into naive C57BL/6 recipients (CD45.2 + ), which were then infected with LM-OVA and engrafted with MC38 cells on day 60 post infection. On days 7–12 after tumor engraftment, recipients were administered PBS, Ad5-WT, or Ad5-OVA daily via the intratumoral route. On day 15 post-tumor engraftment, OT-II CD4 + T cells in various organs were analyzed. (I and J) Frequency (I) and number (J) of OT-II CD4 + cells in the indicated organs of groups administrated PBS (indicated by gray dots), Ad5-WT (indicated by blue dots), or Ad5-OVA (indicated by red dots). All data are representative of at least two independent experiments with at least four mice per group. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by one-way ANOVA with Turkey’s test (B, C, F, G, I, J). Center values and error bars (B, C, F, G, I, J) indicate mean and SEM.

Article Snippet: For the analysis of endogenous CD4 + T cells, lymphocytes were stained with MHC-II LCMV GP 66-77 tetramer (ProImmune, TT3023-4 A) in an incubator at 37°C for 2 h prior to surface marker staining.

Techniques: Infection, Flow Cytometry

Journal: Molecular Therapy Oncology

Article Title: Oncolytic virotherapy mobilizes tumor-resident, granzyme B-producing bystander CD4 + T cells to inhibit systemic microbial infection

doi: 10.1016/j.omton.2026.201187

Figure Lengend Snippet: OV-BYTE-boosted CD4 + T cells are destined for newly differentiated TCF-1 lo CD39 hi differentiation in the periphery (A) UMAP analysis of SM CD4 + cells upon PBS (left, 2,616 cells), NDV-WT (middle, 1,346 cells), or NDV-GP (right, 2,570 cells) treatment in tumors, separated by group. (B) Bar chart of proportions of each UMAP clusters shown in (A). Cell counts (n) for clusters 1–3 in each group are listed below. PBS group: cluster 1 ( n = 606), cluster 2 ( n = 1849), and cluster 3 ( n = 161); NDV-WT group: cluster 1 ( n = 252), cluster 2 ( n = 995), and cluster 3 ( n = 99); NDV-GP group: cluster 1 ( n = 165), cluster 2 ( n = 352), and cluster 3 ( n = 2053). (C) Volcano plot showing differentially expressed genes (DEGs) between cluster 3 and clusters 1 and 2 in (A). The plot displays both up-regulated and down-regulated genes, with the counts of each category provided to the right of the plot. (D) UMAP analysis of SM CD4 + cells upon PBS (navy blue dots, 6,230 cells), NDV-WT (yellow dots, 4,780 cells), or NDV-GP (red dots, 8,591 cells) treatment in spleens. (E) High (≥1,000, red dots) and low (≤1,000, gray dots) expression levels of the top 60 DEGs from (C) shown in the spleen UMAP plot, separated by group. (F) UMAP analysis of SM CD4 + cells upon PBS (navy blue dots, 2,171 cells), NDV-WT (yellow dots, 1,684 cells), or NDV-GP (red dots, 7,416 cells) treatment in lungs. (G) High (≥1,000, red dots) and low (≤1,000, gray dots) expression levels of the top 60 DEGs from (C) shown in the lung UMAP plot, separated by group. (H) Flow cytometry analysis of TCF-1/CD39 expression in SM CD4 + T cells from the PBS-, NDV-WT-, or NDV-GP-treated groups in the indicated organs. Numbers adjacent to the outlined areas indicate percentages of TCF-1 hi CD39 lo cells (blue) or TCF-1 lo CD39 hi cells (red) among SM CD4 + T cells. (I) Frequency of TCF-1 hi CD39 lo (blue) or TCF-1 lo CD39 hi (red) cells among total SM CD4 + T cells from the indicated organs of PBS-, NDV-WT-, and NDV-GP-treated groups. (J) Frequency of TCF-1 hi CD39 lo (blue) or TCF-1 lo CD39 hi (red) cells among endogenous LCMV GP 66-77 tetramer + CD4 + T cells from the indicated organs of PBS-, NDV-WT-, and NDV-GP-treated groups. (K) Frequency of TCF-1 hi CD39 lo (blue) or TCF-1 lo CD39 hi (red) cells among total OT-II CD4 + T cells from the indicated organs of PBS-, Ad5-WT-, and Ad5-OVA-treated groups. All data are representative of at least two independent experiments with at least five mice per group. The Wilcoxon signed-rank test was used in (C). ∗ p < 0.05 and ∗∗∗∗ p < 0.0001 by one-way ANOVA with Turkey’s test (I–K). Center values and error bars (I–K) indicate mean and SEM.

Article Snippet: For the analysis of endogenous CD4 + T cells, lymphocytes were stained with MHC-II LCMV GP 66-77 tetramer (ProImmune, TT3023-4 A) in an incubator at 37°C for 2 h prior to surface marker staining.

Techniques: Expressing, Flow Cytometry

Journal: Molecular Therapy Oncology

Article Title: Oncolytic virotherapy mobilizes tumor-resident, granzyme B-producing bystander CD4 + T cells to inhibit systemic microbial infection

doi: 10.1016/j.omton.2026.201187

Figure Lengend Snippet: OV-BYTE-educated systemic TCF-1 lo CD39 hi CD4 + T cells are imprinted with enhanced effector functionality (A) Venn diagram showing overlap of up-regulated genes in NDV-GP-educated SM CD4 + T cells across different organs. (B) GSEA results showing the normalized enrichment scores (NES) for selected biological process gene sets based on the overlap of 299 DEGs across samples in (A). (C) Heatmap showing selected genes in tumor, spleen, and lung from the indicated groups. (D) Flow cytometry analysis of Ki67 + cells and IFN-γ + TNF-ɑ + cells in indicated SM CD4 + T cells from spleens of indicated groups on day 15 post-tumor engraftment. Numbers adjacent to the outlined areas indicate percentages of Ki67 + cells and IFN-γ + TNF-ɑ + cells among the indicated SM CD4 + T cells. (E and F) Frequency of Ki67 + cells (E) and IFN-γ + TNF-ɑ + cells (F) in PBS-treated TCF1 hi CD39 lo (indicated by gray dots), NDV-WT-treated TCF1 hi CD39 lo (indicated by blue dots), NDV-GP-treated TCF1 hi CD39 lo (indicated by red dots), or NDV-GP-treated TCF1 lo CD39 hi (indicated by purple dots) SM CD4 + T cells from spleens. (G) Flow cytometry analysis of GzmB expression in indicated SM CD4 + T cell populations in spleens. Percentages of GzmB + cells in the indicated SM CD4 + T populations are shown at the top of the flow charts. (H) Frequency of GzmB + cells in PBS-treated TCF1 hi CD39 lo (indicated by gray dots), NDV-WT-treated TCF1 hi CD39 lo (indicated by blue dots), NDV-GP-treated TCF1 hi CD39 lo (indicated by red dots), and NDV-GP-treated TCF1 lo CD39 hi (indicated by purple dots) SM CD4 + T cells in spleens. (I) Schematic of the experimental design. Naive C57BL/6 mice and SM CD4 + T cell chimera C57BL/6 mice were infected with LCMV Armstrong and engrafted with MC38 cells on day 60 post infection. On day 10 after tumor engraftment, splenic Ly108 hi CD39 lo or Ly108 lo CD39 hi SM cells from SM CD4 + T cells chimera recipients were isolated, CTV labeled, and transferred into the other LCMV Armstrong memory cohort via the intravenous route. On days 11–13 after tumor engraftment, recipients were administered NDV-GP daily via the intratumoral route. On day 15 post-tumor engraftment, transferred CTV-labelled SM CD4 + T cells in the spleen were analyzed. (J) Flow cytometry analysis of proliferating cells in the indicated SM CD4 + T cell populations in spleens. Numbers adjacent to the outlined areas indicate percentages of proliferating cells among the indicated SM CD4 + T cells. (K) Frequency of proliferating cells among transferred NDV-GP-treated TCF-1 hi CD39 lo (indicated by red dots) and TCF-1 lo CD39 hi (indicated by purple dots) SM CD4 + T cells from spleens. (L) Schematic of the experimental design. Congenic CD45.1 + SM CD4 + T cells were adoptively transferred into naive C57BL/6 recipients (CD45.2 + ), which were then infected with LCMV Armstrong and engrafted with MC38 cells on day 60 post infection. On days 7–12 after tumor engraftment, recipients were administered NDV-GP daily via the intratumoral route. On day 15 post-tumor engraftment, Ly108 hi CD39 lo or Ly108 lo CD39 hi SM cells in spleens were sorted, and naive SM cells were isolated as a control. Then, Ly108 hi CD39 lo , Ly108 lo CD39 hi , or naive SM cells were in vitro co-cultured with violet hi -labelled LCMV GP 61-80 peptide-coated splenocytes and violet lo -labelled uncoated splenocytes. After 5 h, violet-labeled splenocytes were analyzed. (M) Flow cytometry analysis of violet hi -labelled LCMV GP 61-80 peptide-coated cells and violet lo -labeled uncoated control cells among live target cells in the presence of naive, TCF-1 hi CD39 lo , or TCF-1 lo CD39 hi SM cells. Numbers adjacent to the red outlined areas indicate percentages of LCMV GP 61-80 peptide-coated cells, and numbers adjacent to the blue outlines indicate percentages of uncoated control cells. (N) Ex vivo killing efficacy of TCF-1 hi CD39 lo (indicated by red dots) and TCF-1 lo CD39 hi SM cells (indicated by purple dots). All data are representative of at least two independent experiments with at least three mice per group. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.0001 by one-way ANOVA with Turkey’s test (E, F, H) and two-tailed unpaired Student’s t test (K, N). Center values and error bars (E, F, H, K, N) indicate mean and SEM.

Article Snippet: For the analysis of endogenous CD4 + T cells, lymphocytes were stained with MHC-II LCMV GP 66-77 tetramer (ProImmune, TT3023-4 A) in an incubator at 37°C for 2 h prior to surface marker staining.

Techniques: Flow Cytometry, Expressing, Infection, Isolation, Labeling, Control, In Vitro, Cell Culture, Ex Vivo, Two Tailed Test

Journal: Molecular Therapy Oncology

Article Title: Oncolytic virotherapy mobilizes tumor-resident, granzyme B-producing bystander CD4 + T cells to inhibit systemic microbial infection

doi: 10.1016/j.omton.2026.201187

Figure Lengend Snippet: Systemic TCF-1 lo CD39 hi CD4 + T cells are more prone to T H 1 differentiation upon OV-BYTE treatment (A) Bubble chart showing CD4 + T cell lineage-associated key genes in Tcf7 lo Entpd1 hi clusters and Tcf7 hi Entpd1 lo clusters in tumor, spleen, and lung. (B) Schematic diagram of the experimental design. Congenic CD45.1 + CXCR5-GFP SM CD4 + T cells were adoptively transferred into naive C57BL/6 recipients (CD45.2 + ), which were then infected with LCMV Armstrong and engrafted with MC38 cells on day 60 post infection. On days 7–12 after tumor engraftment, recipients were administered PBS, NDV-WT, or NDV-GP daily via the intratumoral route. On day 15 post-tumor engraftment, CXCR5-GFP SM CD4 + T cells from tumor, spleen, and lung were analyzed. (C) Flow cytometry analysis of CXCR5/Ly6C expressions in the indicated SM CD4 + T cell populations in the indicated organs. (D) Frequency of CXCR5/Ly6C expressions in the indicated SM CD4 + T cell populations from tumor, spleen, and lung. (E) Flow cytometry analysis of T-bet expression levels in the indicated SM CD4 + T cell populations from tumor, spleen, and lung. Numbers in the flow charts indicate T-bet MFI of PBS-treated TCF-1 hi CD39 lo (gray), NDV-WT-treated TCF-1 hi CD39 lo (blue), NDV-GP-treated TCF-1 hi CD39 lo (red), and NDV-GP-treated TCF-1 lo CD39 hi (purple) SM CD4 + T cells. (F) Comparison of T-bet expression levels among the indicated SM CD4 + T cell populations from tumor, spleen, and lung. All data are representative of at least two independent experiments with at least four mice per group. Not significant (ns), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by one-way ANOVA with Turkey’s test (D, F). Center values and error bars (D, F) indicate mean and SEM.

Article Snippet: For the analysis of endogenous CD4 + T cells, lymphocytes were stained with MHC-II LCMV GP 66-77 tetramer (ProImmune, TT3023-4 A) in an incubator at 37°C for 2 h prior to surface marker staining.

Techniques: Infection, Flow Cytometry, Expressing, Comparison

Journal: Molecular Therapy Oncology

Article Title: Oncolytic virotherapy mobilizes tumor-resident, granzyme B-producing bystander CD4 + T cells to inhibit systemic microbial infection

doi: 10.1016/j.omton.2026.201187

Figure Lengend Snippet: OV-BYTE-educated systemic TCF-1 lo CD39 hi CD4 + T cells epigenetically resemble tumor-resident CD4 + T BYS cells (A) Schematic of the experimental design for (B–F). Congenic CD45.1 + SM CD4 + T cells were adoptively transferred into naive C57BL/6 recipients (CD45.2 + ), which were then infected with LCMV Armstrong and engrafted with MC38 cells on day 60 post infection. On days 7–12 after tumor engraftment, recipients were administered PBS, NDV-WT, or NDV-GP daily via intratumoral route. On day 15 post-tumor engraftment, PBS-treated Ly108 hi CD39 lo , NDV-WT-treated Ly108 hi CD39 lo , NDV-GP-treated Ly108 hi CD39 lo , and NDV-GP-treated Ly108 lo CD39 hi SM CD4 + T cells from tumors and spleens were isolated and subjected to ATAC-seq. (B) PCA analysis of peak accessibility in sorted cells from (A). The plot shows the first two principal components (PC1 and PC2), which explain 71.3% and 14.4% of the total variance, respectively. Each group includes two biological replicates from two independent experiments. (C) UpSet plot illustrating uniquely open DARs in each group, as well as the overlap of open DARs between each pair of groups. (D) Venn diagram showing the overlap of open DARs among the indicated groups. The numbers and percentages above each section represent the overlap of DARs for the respective groups. (E) Venn diagram showing the overlap of open DARs among the indicated groups. These open DARs correspond to the top 60 up-regulated DEGs in the tumor TCF-1 lo CD39 hi subset. The numbers above each section represent the overlap of DARs for the respective groups. (F) ATAC-seq signal profiles of the indicated gene loci. Differential peaks are highlighted in red frames.

Article Snippet: For the analysis of endogenous CD4 + T cells, lymphocytes were stained with MHC-II LCMV GP 66-77 tetramer (ProImmune, TT3023-4 A) in an incubator at 37°C for 2 h prior to surface marker staining.

Techniques: Infection, Isolation

Journal: Molecular Therapy Oncology

Article Title: Oncolytic virotherapy mobilizes tumor-resident, granzyme B-producing bystander CD4 + T cells to inhibit systemic microbial infection

doi: 10.1016/j.omton.2026.201187

Figure Lengend Snippet: Tumor-resident CD4 + T BYS cells serve as the reservoir of OV-BYTE-induced systemic TCF-1 lo CD39 hi CD4 + T cells (A) Schematic of the experimental design for (B–H). Congenic CD45.1 + CD45.2 - SM CD4 + T cells were adoptively transferred into naive C57BL/6 recipients (CD45.2 + ), which were then infected with LCMV Armstrong. In another cohort, naive C57BL/6 recipients (CD45.2 + ) receiving congenic CD45.1 + CD45.2 + SM CD4 + T cells were then infected with LCMV Armstrong. On day 60 post-infection, recipients that received CD45.1 + CD45.2 - SM CD4 + T cells were engrafted with MC38 cells. On day 10 post-tumor growth, tumor masses were excised and transplanted into recipients that received CD45.1 + CD45.2 + SM CD4 + T cells. The transplanted tumors were then subjected to intratumoral injections of PBS, NDV-WT, or NDV-GP, and flow cytometry analysis was performed at the indicated time point. (B) Flow cytometry analysis of CD45.1 + CD45.2 - and CD45.1 + CD45.2 + SM CD4 + T cells in total SM CD4 + T cells from the indicated organs of PBS-, NDV-WT-, or NDV-GP-treated group. Numbers adjacent to the red outlined areas indicate the percentages of CD45.1 + CD45.2 - SM CD4 + T cells (donor), and numbers adjacent to the blue ones indicate the percentage of CD45.1 + CD45.2 + SM CD4 + T cells (host). (C and D) Frequency (C) and number (D) of CD45.1 + CD45.2 - SM CD4 + T cells (donor) from the indicated organs of group treated with PBS (indicated by gray dots), NDV-WT (indicated by blue dots), or NDV-GP (indicated by red dots). Quantification of SM CD4 + cells from 100 μL of peripheral blood. (E and F) Frequency of TCF-1 hi CD39 lo (E) and TCF-1 lo CD39 hi (F) cells among host-derived PBS-treated (indicated by gray dots), NDV-WT-treated (indicated by blue dots), NDV-GP-treated (indicated by red dots) SM CD4 + T cells, and donor-derived NDV-GP-treated (indicated by purple dots) SM CD4 + T cells in the indicated organs. (G) Frequency of circulating cells (intravenous [i.v.] CD4 staining positive) of host-derived PBS-treated (indicated by gray dots), NDV-WT-treated (indicated by blue dots), NDV-GP-treated (indicated by red dots) TCF-1 hi CD39 lo SM CD4 + T cells, and NDV-GP-treated TCF-1 lo CD39 hi (indicated by purple dots) SM CD4 + T cells, together with donor-derived NDV-GP-treated TCF-1 hi CD39 lo (indicated by red cubes) and TCF-1 lo CD39 hi (indicated by purple cubes) SM CD4 + T cells in the indicated organs. (H) Comparison of GzmB expression levels in host-derived PBS-treated (indicated by gray dots), NDV-WT-treated (indicated by blue dots), NDV-GP-treated (indicated by red dots) TCF-1 hi CD39 lo SM CD4 + T cells, and NDV-GP-treated TCF-1 lo CD39 hi (indicated by purple dots) SM CD4 + T cells, together with donor-derived NDV-GP-treated TCF-1 hi CD39 lo (indicated by red cubes) and TCF-1 lo CD39 hi (indicated by purple cubes) SM CD4 + T cells in the indicated organs. All data are representative of at least two independent experiments with at least four mice per group. Not significant (ns), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by one-way ANOVA with Turkey’s test (C–H). Center values and error bars (C–H) indicate mean and SEM.

Article Snippet: For the analysis of endogenous CD4 + T cells, lymphocytes were stained with MHC-II LCMV GP 66-77 tetramer (ProImmune, TT3023-4 A) in an incubator at 37°C for 2 h prior to surface marker staining.

Techniques: Infection, Flow Cytometry, Derivative Assay, Staining, Comparison, Expressing

Journal: Molecular Therapy Oncology

Article Title: Oncolytic virotherapy mobilizes tumor-resident, granzyme B-producing bystander CD4 + T cells to inhibit systemic microbial infection

doi: 10.1016/j.omton.2026.201187

Figure Lengend Snippet: Turnover of OV-BYTE-educated Ly108 hi CD39 lo and Ly108 lo CD39 hi CD4 + T cells (A) Schematic of the experimental design for (B–I). Congenic CD45.1 + SM CD4 + T cells were adoptively transferred into naive C57BL/6 recipients (CD45.2 + ), which were then infected with LCMV Armstrong. In another cohort, naive C57BL/6 recipients (CD45.2 + ) were simultaneously infected with LCMV Armstrong without SM CD4 + T cell transfer. On day 60 post-infection, both cohorts were engrafted with MC38 cells. On days 7–12 post-tumor engraftment, the cohort that received SM CD4 + T cells was administered NDV-GP daily via the intratumoral route. Then, Ly108 hi CD39 lo and Ly108 lo CD39 hi SM CD4 + T cells in tumor and spleen were isolated on day 15 post-tumor engraftment and subsequently transferred into MC38 tumor-bearing mice (no SM CD4 + T cells transfer) via intratumoral injection and intravenous injection, respectively. These recipients were then administered NDV-GP intratumorally for five consecutive days, and flow cytometry analysis was thereafter. (B and C) Frequency (B) and number (C) of transferred tumor-derived Ly108 hi CD39 lo (indicated by blue dots) and Ly108 lo CD39 hi (indicated by red dots) SM CD4 + T cells in the indicated organs. Quantification of SM CD4 + cells from 100 μL of peripheral blood. (D) Flow cytometry analysis of TCF-1 hi CD39 lo and TCF-1 lo CD39 hi cells in transferred tumor-derived Ly108 hi CD39 lo or Ly108 lo CD39 hi SM CD4 + T cells (red contour lines) in the indicated organs. TCF-1/CD39 expression of tumor-resident polyclonal CD4 + T cells (gray contour lines) is shown as an internal control in the bottom-left plot. Numbers adjacent to the blue outlined areas indicate the percentages of TCF-1 hi CD39 lo cells in the indicated transferred SM CD4 + T cells, and numbers adjacent to the red ones indicate the percentage of TCF-1 lo CD39 hi cells in the indicated transferred SM CD4 + T cells. (E and F) Frequency of TCF-1 lo CD39 hi (E) and TCF-1 hi CD39 lo (F) cells in transferred tumor-derived Ly108 hi CD39 lo (indicated by blue dots) and Ly108 lo CD39 hi (indicated by red dots) SM CD4 + T cells in the indicated organs. (G and H) Frequency (G) and number (H) of transferred spleen-derived Ly108 hi CD39 lo (indicated by blue dots) and Ly108 lo CD39 hi (indicated by red dots) SM CD4 + T cells in the indicated organs. Quantification of SM CD4 + cells from 100 μL of peripheral blood. (I) Frequency of TCF-1 hi CD39 lo and TCF-1 lo CD39 hi cells in transferred spleen-derived Ly108 lo CD39 hi SM CD4 + T cells. All data are representative of at least two independent experiments with at least four mice per group. Not significant (ns), ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 by two-tailed unpaired Student’s t test (B, C, E–H). Center values and error bars (B, C, E–H) indicate mean and SEM.

Article Snippet: For the analysis of endogenous CD4 + T cells, lymphocytes were stained with MHC-II LCMV GP 66-77 tetramer (ProImmune, TT3023-4 A) in an incubator at 37°C for 2 h prior to surface marker staining.

Techniques: Infection, Isolation, Injection, Flow Cytometry, Derivative Assay, Expressing, Control, Two Tailed Test

Journal: Molecular Therapy Oncology

Article Title: Oncolytic virotherapy mobilizes tumor-resident, granzyme B-producing bystander CD4 + T cells to inhibit systemic microbial infection

doi: 10.1016/j.omton.2026.201187

Figure Lengend Snippet: Dual protection against tumor and pathogen infection by the OV-BYTE strategy (A) Schematic of the experimental design for (B–D). C57BL/6 mice were infected with LCMV Armstrong and engrafted with MC38 cells on day 60 post-infection. On days 7–12 after tumor engraftment, recipients were daily administered PBS, NDV-WT, or NDV-GP daily via the intratumoral route. On day 15 after tumor engraftment, recipients were infected with either LM-GP 61-80 or IAV-GP 61-80 at an LD 50 dose. (B) Tumor growth curve of MC38 tumor-bearing mice intratumorally treated with PBS, NDV-WT, or NDV-GP as described in (A). (C and D) Survival curves of LM-GP 61-80 (C) and IAV-GP 61-80 (D) infection in MC38-engrafted mice treated with PBS, NDV-WT, or NDV-GP as described in (A). (E) Schematic of the experimental design for (F–H). C57BL/6 mice were infected with LCMV Armstrong and engrafted with B16F10 cells on day 60 post-infection. On days 7–12 after tumor engraftment, recipients were administered PBS, Ad5-WT, or Ad5-GP daily via the intratumoral route. On day 15 after tumor engraftment, recipients were infected with either LM-GP 61-80 or IAV-GP 61-80 at an LD 50 dose. (F) Tumor growth curve of B16F10 tumor-bearing mice intratumorally treated with PBS, Ad5-WT, or Ad5-GP as described in (E). (G and H) Survival curves of LM-GP 61-80 (G) and IAV-GP 61-80 (H) infection in B16F10-engrafted mice treated with PBS, Ad5-WT, or Ad5-GP as described in (E). (I) Schematic of the experimental design. Congenic CD45.1 + SM CD4 + T cells were adoptively transferred into naive C57BL/6 recipients (CD45.2 + ), which were then infected with LCMV Armstrong. On day 60 post-infection, these recipients were engrafted with MC38 cells. On days 7–12, these recipients were administered NDV-GP daily via the intratumoral route. Then, Ly108 hi CD39 lo and Ly108 lo CD39 hi SM CD4 + T cells in the spleens were isolated on day 15 post-tumor engraftment and subsequently transferred into MC38 tumor-bearing mice (no LCMV Armstrong infection) via intravenous injection, along with MC38 tumor-bearing mice receiving no cell transfer as control. One day later, all recipients were infected with LM-GP 61-80 at an LD 50 dose. (J) Survival curve of LM-GP 61-80 infection in groups described in (I). (K) Schematic of the experimental design. WT and Gzmb KO mice were infected with LCMV Armstrong. On day 60 post-infection, splenic LCMV Armstrong-activated CD4 + T MEM cells were harvested and adoptively transferred into another cohort of naive C57BL/6 mice. These recipients, along with control C57BL/6 mice with no CD4 + T MEM cell transfer, were then engrafted with MC38 tumor cells, intratumorally administrated NDV-WT or NDV-GP, and infected with LM-GP 61-80 at the indicated time points. (L) Survival curve of LM-GP 61-80 infection in groups described in (I). All data are representative of at least two independent experiments with at least eight mice per group. Not significant (ns), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by two-way ANOVA (B, F) and log rank (Mantel-Cox) test (C, D, G, H, J, L). Center values and error bars (B, F) indicate mean and SEM.

Article Snippet: For the analysis of endogenous CD4 + T cells, lymphocytes were stained with MHC-II LCMV GP 66-77 tetramer (ProImmune, TT3023-4 A) in an incubator at 37°C for 2 h prior to surface marker staining.

Techniques: Infection, Isolation, Injection, Control