vivo | anti mouse cd8a  (Bio X Cell)

Journal: Cell reports

Article Title: Semaphorin7A and PD-L1 cooperatively drive immunosuppression during mammary involution and breast cancer

doi: 10.1016/j.celrep.2025.115676

Figure Lengend Snippet: (A) Percent SEMA7A + tumor cells isolated from E0771 control (Ctrl) and SEMA7A overexpressing (S7A OE) tumors. (B) Geometric mean fluorescence intensity (gMFI) of SEMA7A expression from Ctrl or S7A OE tumors. (C) Percent PD-L1+ tumor cells from Ctrl or S7A OE tumors. (D) Percent PD-L1+ immune cells from Ctrl or S7A OE tumors. (E) Percent PD-L1+ total macrophages from Ctrl or S7A OE tumors. (F) Percentage PD-L1+ M1-like macrophages, M2-like macrophages, and PoEMs from Ctrl or S7A OE tumors. (G) Frequency of CD4 + and CD8 + T cells of total immune cells from Ctrl or S7A OE tumors. (H) gMFI of PD-1 on CD4 + and CD8 + T cells from Ctrl or S7A OE tumors. (I) Percent exhausted (PD-1 high /LAG-3+/IFNγ–/TNFα–) CD4 + and CD8 + T cells of total CD4/8 T cells from Ctrl or S7A OE tumors. (J) Percent activated (PD-1 mid /LAG-3–/IFNγ+/TNFα+) CD4 + and CD8 + T cells of total CD4/8 T cells from Ctrl or S7A OE tumors. (K) Summary of macrophage and T cell phenotypes between SEMA7A Low and SEMA7A High tumors. Error bars are mean ± SEM. Sample sizes for (A)–(J): Ctrl n = 5 tumors; S7A OE n = 5 tumors. Statistics for (A)–(J) are unpaired two-tailed Welch’s t tests. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: In vivo | Anti-mouse CD8a (Clone 53-6.7) , Bio X Cell , Cat# BE0004-1; RRID: AB_1107671.

Techniques: Isolation, Control, Fluorescence, Expressing, Two Tailed Test

Journal: Cell reports

Article Title: Semaphorin7A and PD-L1 cooperatively drive immunosuppression during mammary involution and breast cancer

doi: 10.1016/j.celrep.2025.115676

Figure Lengend Snippet: (A) Percent CD4 + T cells from E0771 control (Ctrl) or SEMA7A overexpressing (OE) tumors following treatment with IgG control, αPD-1, or αPD-L1. (B) Percent CD8 + T cells from Ctrl or OE tumors following treatment with IgG control, αPD-1, or αPD-L1. (C) PD-1 gMFI on CD4 + T cells from Ctrl or S7A OE tumors following treatment with IgG control, αPD-1, or αPD-L1. (D) PD-1gMFI on CD8 + T cells from Ctrl or S7A OE tumors following treatment with IgG control, αPD-1, or αPD-L1. (E) Percent exhausted (PD-1 high /LAG-3+/IFNγ–/TNFα–) CD4 + T cells from Ctrl or S7A OE tumors following treatment with IgG control, αPD-1, or αPD-L1. (F) Percent exhausted (PD-1 high /LAG-3+/IFNγ–/TNFα–) CD8 + T cells from Ctrl or S7A OE tumors following treatment with IgG control, αPD-1, or αPD-L1. (G) Percent activated (PD-1 mid /LAG-3-/IFNγ+/TNFα+) CD4 + T cells from Ctrl or S7A OE tumors following treatment with IgG control, αPD-1, or αPD-L1. (H) Percent activated (PD-1 mid /LAG-3-/IFNγ+/TNFα+) CD8 + T cells from Ctrl or S7A OE tumors following treatment with IgG control, αPD-1, or αPD-L1. (I) Growth of E0771 OE tumors. Arrows indicate administration of treatment with IgG control, αPD-1, or αPD-L1. IgG n = 6 tumors; αPD-1 n = 6 tumors; αPD-L1 n = 8 tumors. (J) Growth of E0771 OE tumors in CD4 + T cell-depleted mice with/without αPD-1/αPD-L1 treatment. αCD4 was administered at days 6, 8, 13, and 18. Arrows represent IgG, αPD-1, or αPD-L1 treatments. αCD4 + IgG n = 6 tumors; αCD4 + αPD-1 n = 6 tumors; αCD4 + αPD-L1 n = 8 tumors. (K) Growth of E0771 OE tumors in CD8 + T cell-depleted mice with/without αPD-1/αPD-L1 treatment. αCD8 was administered at days 6, 8, 13, and 18. Arrows represent IgG, αPD-1, or αPD-L1 treatments. αCD8 + IgG n = 6 tumors; αCD8 + αPD-1 n = 6 tumors; αCD8 + αPD-L1 n = 8 tumors. Error bars are mean ± SEM. Sample sizes for (A)–(H): Ctrl + IgG n = 6 tumors; Ctrl + αPD-1 n = 8 tumors; Ctrl + αPD-L1 n = 6 tumors; OE + IgG n = 6 tumors; OE + αPD-1 n = 8 tumors; OE + αPD-L1 n = 6 tumors. Statistics for (A)–(H) are ordinary one-way ANOVA with Tukey’s multiple comparisons test. Statistics for (I)–(K) are from day 20 using ordinary one-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. # indicates statistically significant differences from all other groups .

Article Snippet: In vivo | Anti-mouse CD8a (Clone 53-6.7) , Bio X Cell , Cat# BE0004-1; RRID: AB_1107671.

Techniques: Control

Journal: Cell reports

Article Title: Semaphorin7A and PD-L1 cooperatively drive immunosuppression during mammary involution and breast cancer

doi: 10.1016/j.celrep.2025.115676

Figure Lengend Snippet: (A) Growth of individual E0771 SEMA7A overexpressing (OE) tumors following treatment with IgG control ( n = 10 tumors) or αSEMA7A (SmAbH1) ( n = 10 tumors). (B) Survival analysis from (A). n = 5 mice/group with Mantel-Cox test. (C) Response rates of individual tumors from (A). n = 10 tumors/group. (D) Growth of 66cL4 OE tumors following treatment with IgG control ( n = 12 tumors), SmAbH1 ( n = 12 tumors), or aPD-L1 ( n = 12 tumors). Six of six OE + IgG and six of six OE + aPD-L1 mice were euthanized at day 33 due to tumor burden or morbidity, three of six OE + SmAbH1 mice were euthanized for time-matched flow cytometry. Statistics are from day 33 tumor volumes using ordinary one-way ANOVA with Tukey’s multiple comparisons test. (E) Growth rates from (D). (F) Response rates of individual tumors from (D). n = 12 tumors/group. (G) Percent immune cells from tumors following treatments in (D). (H) Percent PD-L1+ total macrophages from tumors following treatments in (D). (I) Percent PD-L1+ M2-like macrophages from tumors following treatments in (D). (J) Percent PD-L1+ PoEMs from tumors following treatments in (D). (K) Percent CD4 + T cells from tumors following treatments in (D). (L) PD-1 geometric mean fluorescence intensity (gMFI) on CD4 + T cells from tumors following treatments in (D). (M) Percent exhausted (PD-1 high /LAG-3+/IFNγ–/TNFα–) CD4 + T cells from tumors following treatments in (D). (N) Percent activated (PD-1 mid /LAG-3–/IFNγ+/TNFα+) CD4 + T cells from tumors following treatments in (D). (O) Percent CD8 + T cells from tumors following treatments in (D). (P) PD-1 gMFI on CD8 + T cells from tumors following treatments in (D). (Q) Percent exhausted (PD-1 high /LAG-3+/IFNγ–/TNFα–) CD8 + T cells from tumors following treatments in (D). (R) Percent activated (PD-1 mid /LAG-3-/IFNγ+/TNFα+) CD8 + T cells from tumors following treatments in (D). Error bars are mean ± SEM. Sample sizes for (G)–(R): n = 5 tumors/group. Statistics for (G)–(R) are ordinary one-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: In vivo | Anti-mouse CD8a (Clone 53-6.7) , Bio X Cell , Cat# BE0004-1; RRID: AB_1107671.

Techniques: Control, Flow Cytometry, Fluorescence

Journal: Cell Death & Disease

Article Title: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

doi: 10.1038/s41419-025-07570-8

Figure Lengend Snippet: a The protein levels of Smyd3 and Shcbp1 in normal mouse mammary glands, and tumors initiated with HP5712 cells in 3 months old FVB virgin mice as shown by Western blots. b The protein levels of Smyd3 and Shcbp1 in HP5712 cells expressing sgSmyd3, OE-Smyd3, or OE-Smyd3/sgShcbp1 by Western blots. Representative tumor images ( c ) and tumor weight plots ( d ) at day 32 from FVB virgin mice implanted with HP5712 parental (HP tumors), OE-Smyd3-HP5712, sgSmyd3-HP5712, and OE-Smyd3/sgShcbp1-HP5712 cells at 1X10 6 cells per mammary fat pad (n = 8 mice/group). e The plot of relative spleen weight from the same cohort of mice in c and d . f The protein levels of Smyd3, H3K4me3, and Shcbp1 in tumors initiated with HP5712 parental, OE-Smyd3-HP5712, sgSmyd3-HP5712, and OE-Smyd3/sgShcbp1-HP5712 cells by Western blots. g The protein levels of pMek, pErk, and Kras in tumors initiated with HP5712 parental, OE-Smyd3-HP5712, sgSmyd3-HP5712, and OE-Smyd3/sgShcbp1-HP5712 cells as shown by Western blots. h tSNE visualized immune cells from normal FVB mouse mammary glands (FVB MG, n = 6 mice), tumors implanted with parental HP5712 cells (HP Vector Ctr), HP-OE-Smyd3, HP-OE-Smyd3/sgShcbp1, and HP-sgSmyd3 from FVB mice (n = 6 mice/group) by CyTOF analysis. Quantifications of PMN-MDSCs and M-MDSCs ( i ), CD4+ and CD8 + T cells ( j ) in CD45+ immune cell populations from the same cohorts of mice in h (n = 3 biological independent samples/per group). k , l The WT-T cells were activated with CD3 and CD28 antibody. Co-cultured the activated T cells without or with MDSCs from the mice implanted with HP parental cells, HP cells expressing Smyd3, or sgSmyd3, or OE-Smyd3/sgShcbp1 for 72 hours and examined T cells proliferation status ( k ) and quantifications ( l ) of data in k (n = 3 mice/group). Representative images of CD8+/PD1+ double positive cells in mammary tissues from the mice with the genotypes of HP-Ctr, HP-OE-Smyd3, HP-sgSmyd3, and HP-OE-Smyd3/sgShcbp1 ( m ) and quantifications of CD8+/PD1 double positive cells from mammary tissues ( n ) in these mice (n = 4 mice/group, and at least 20 images for each sample were counted). Scale bar: 20 μm. arrow heads point to PD1/CD8 positive cell. o Expressions of PD1 from the spleen with implantation of parental HP cells, HP cells with OE-Smyd3, HP-sgSmyd3, and HP-OE-Smyd3/sgShcbp1 in FVB mice as determined by qPCR. p Summary of Smyd3-Shcbp1oncogenic signals shape the TIME.

Article Snippet: In Vivo anti-mouse CD8a , BioX Cell , BP0117 , , .

Techniques: Western Blot, Expressing, Plasmid Preparation, Cell Culture

Journal: Cell Death & Disease

Article Title: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

doi: 10.1038/s41419-025-07570-8

Figure Lengend Snippet: Representative tumor images ( a ), and tumor weight plots ( b ) at day 22 from FVB mice implanted with HP5712 cells at 1×10 6 cells per mammary fat pad, and treatment with PBS, PD1, trametinib (Tra), and PD1+Tra (n = 8 mice/group). c The plot of tumor volume in the processes of treatment in ( a , b ). d The plot of relative spleen weight from the same cohort of mice in a , b . Quantifications of PMN-MDSCs and M-MDSCs ( e ), CD4+ and CD8 + T cells ( f ) in CD45+ immune cell populations from the same cohorts of mice in a , b by CyTOF analysis at D12 (n = 3 mice/group). Quantifications of PMN-MDSCs and M-MDSCs ( g ), CD4+ and CD8 + T cells ( h ) in CD45+ immune cell populations from the same cohorts of mice in a , b by CyTOF analysis at D22 (n = 3 mice/group). i , j Tumor images from the HP Ctr mice, HP mice treated with αPD1 and Tra, and HP mice treated with αPD1 and Tra with depletion of T cell using CD8 antibody ( i ). The plot of tumor weight ( j ) in the same cohort of mice in ( i ) (n = 8 mice in each group). The protein levels of Smyd3 and Shcbp1 in tumors initiated with HP5712 cells and treatment with PBS, αPD1, Tra, and αPD1+Tra in FVB mice at D12 ( k ) and D22 ( l ) as shown by Western blots. The protein levels of pMek, pErk, Kras, and Grb2 in tumors initiated with HP5712 cells and treatment with PBS, αPD1, Tra, and αPD1+Tra in FVB mice at D12 ( m ) and D22 ( n ) as shown by Western blots. Expressions of Smyd3 ( o ) and Shcbp1 ( p ) in G600 cells with the treatment of E2, Tra, and E2 together with Tra at 1 hour, 2 hours, 4 hours, and 24 hours as determined by qPCR.

Article Snippet: In Vivo anti-mouse CD8a , BioX Cell , BP0117 , , .

Techniques: Western Blot

Journal: Cell Death & Disease

Article Title: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

doi: 10.1038/s41419-025-07570-8

Figure Lengend Snippet: Antibodies used for ChIP, IHC, IF, WB and in vivo mouse study.

Article Snippet: In Vivo anti-mouse CD8a , BioX Cell , BP0117 , , .

Techniques: Immunohistochemistry-IF, In Vivo

Journal: Cancer cell

Article Title: Tailoring vascular phenotype through AAV therapy promotes anti-tumor immunity in glioma.

doi: 10.1016/j.ccell.2023.04.010

Figure Lengend Snippet: Figure 5. CD8+ T cell phenotypes differ based on response after AAV-LIGHT therapy (A) Schematic illustration of the experimental layout used to obtain data shown in (B)–(K). i.c., intracranial. In brief, the murine glioma cell line CT-2A-Ova was implanted intracranially in C57BL/6 mice on day 0, followed by intravenous administration of either AAV-GFP or AAV-LIGHT on days 4, 7, and 10 post-tumor implantation. Mice were sacrificed on days 14 and 21 post-tumor implantation, and the brains were collected for single-cell RNA sequencing (scRNA-seq) analysis or flow cytometry analysis. (B) At the time of collection, brains were dissected sagittally and visually assessed to determine whether a macroscopic tumor was present. Samples were classified accordingly into two categories depending on tumor status: ‘‘tumor’’ (when a tumor was present) or ‘‘no macroscopic tumor’’ (when no clear tumor mass was observed). Pie charts show the distribution of these categories within each treatment group at days 14 and 21. (C) Uniform manifold approximation and projection (UMAP) of scRNA-seq profiles of 7,904 tumor-infiltrating CD8+ T cells. Colors indicate cell distribution de- pending on tumor status (left) or treatment (right). (D) AUCell enrichment scores of gene signatures upregulated in effector vs. exhausted CD8+ T cells (GSE9650), memory vs. exhausted CD8+ T cells (GSE9650), exhausted CD8+ T cells, and stem-like CD8+ T cells.27,28

Article Snippet: In vivo antibody therapies Rat-a-mouse CD8a (BioXCell, Lebanon, NH, clone: 53-6.7, #BE0004) or rat IgG2a isotype control (BioXcell, clone: 2A3, #BE0089) antibodies were administered i.v. every 6 days (200 mg/dose), starting at day -2 from tumor implantation for depleting CD8+ T cells.

Techniques: Tumor Implantation, RNA Sequencing, Cytometry

Journal: Cancer cell

Article Title: Tailoring vascular phenotype through AAV therapy promotes anti-tumor immunity in glioma.

doi: 10.1016/j.ccell.2023.04.010

Figure Lengend Snippet: Figure 6. AAV-LIGHT therapy promotes stem-like CD8+ T cells during an ongoing anti-tumor response (A) Schematic illustration of the experimental layout used to obtain data shown in (B)–(K). In brief, brains with macroscopic CT-2A-Ova tumors were collected on day 14 post-tumor implantation from AAV-GFP- or AAV-LIGHT-treated mice (n = 2 mice/group) for single-cell RNA sequencing (scRNA-seq) analysis. A trajectory

Article Snippet: In vivo antibody therapies Rat-a-mouse CD8a (BioXCell, Lebanon, NH, clone: 53-6.7, #BE0004) or rat IgG2a isotype control (BioXcell, clone: 2A3, #BE0089) antibodies were administered i.v. every 6 days (200 mg/dose), starting at day -2 from tumor implantation for depleting CD8+ T cells.

Techniques: Tumor Implantation, RNA Sequencing

Journal: Cancer cell

Article Title: Tailoring vascular phenotype through AAV therapy promotes anti-tumor immunity in glioma.

doi: 10.1016/j.ccell.2023.04.010

Figure Lengend Snippet: Figure 7. TCF1+CD8+ stem-like cells are present in T cell-rich TLS and within the tumor area in both murine and human gliomas (A and B) Multispectral staining showing the presence of TCF1+CD8+ stem-like cells within TLS in the brain of CT-2A glioma-bearing mice treated with either (A) AAV-GFP or (B) AAV-LIGHT. (C) Multispectral staining showing the presence of TCF1+CD8+ stem-like cells within the tumor area in the brain of CT-2A glioma-bearing mice treated with either AAV-GFP or AAV-LIGHT. In (A)–(C), dotted boxes indicate the areas shown in the zoom images below the main images. White arrows indicate TCF1+CD8+ T cells. Main scale bars, 50 mm; zoom scale bars, 10 mm. (D) Quantification of distance (mm) to the nearest CD11c+ cell from CD8+ or TCF1+CD8+ T cells within the tumor area in the indicated groups (n = 3–4 mice/group). Statistics: Kruskal-Wallis test with Dunn’s correction.

Article Snippet: In vivo antibody therapies Rat-a-mouse CD8a (BioXCell, Lebanon, NH, clone: 53-6.7, #BE0004) or rat IgG2a isotype control (BioXcell, clone: 2A3, #BE0089) antibodies were administered i.v. every 6 days (200 mg/dose), starting at day -2 from tumor implantation for depleting CD8+ T cells.

Techniques: Staining

Journal: Cancer cell

Article Title: Tailoring vascular phenotype through AAV therapy promotes anti-tumor immunity in glioma.

doi: 10.1016/j.ccell.2023.04.010

Figure Lengend Snippet: Figure 8. Delivery of an LTbR-IgG construct abolishes the survival benefit of AAV-LIGHT, in association with reduced formation of TA-HEVs and a reduction in stem-like CD8+ T cells (A) Schematic illustration of the experimental layout used to obtain data shown in (B)–(N). i.c., intracranial; IF, immunofluorescence. In brief, murine glioma cell lines CT-2A or CT-2A-Ova were implanted intracranially in C57BL/6 mice on day 0, followed by intravenous administration of either AAV-GFP or AAV-LIGHT on days 4, 7, and 10 post-tumor implantation. LTbR-IgG construct or control IgG were administered intraperitoneally every 7 days starting at day 4 from tumor implantation. Mice were sacrificed at the humane endpoint, and tumor-bearing brains were collected for immunofluorescence analysis (n = 20 mice/group), or mice were sacrificed on day 14 post-tumor implantation, and tumor-bearing brains were harvested for flow cytometry analysis of tumor-infiltrating T cells (n = 3–7 mice/group). (B) Kaplan-Meier survival curves of CT-2A tumor-bearing mice treated with AAV-LIGHT or AAV-GFP and LTbR-IgG or control IgG (n = 20 mice/group). Statistics: log-rank test. The graph shows the combined survival curves from two independent experiments. (C) Representative immunofluorescence images of CD45+B220+CD3+ TLS in the indicated groups. (D and E) Quantification of (D) the number of TLS per brain (n = 7–8 mice/group) and (E) TLS area (mm2) (n = 16–18 TLS/group) in the indicated groups. (F) Representative immunofluorescence images of CD3+ T cells and B220+ B cells within the same TLS in the indicated groups. (G) Quantification of the CD3-to-B220 ratio within the TLS (n = 16–18 TLS/group) in the indicated groups. (H) Representative immunofluorescence images of MAdCAM-1+CD31+ TA-HEVs and MAdCAM-1CD31+ tumor vessels in the indicated groups. (I) Proportion of CD31+ area that was positive for MAdCAM-1 (n = 5 mice/group) in the indicated groups. (J) Quantification of CD8+ T cells (percentage of CD3+ T cells) in the indicated groups (n = 3–6 mice/group). (K) Representative pseudocolor density plots of PD-1 vs. TCF1 expression on Ovadex+CD8+ T cells in the indicated groups. (L–N) Quantification of (L) TCF1+PD-1+ cells (percentage of Ovadex+CD8+ cells), (M) TCF1PD-1+ cells (percentage of Ovadex+CD8+ cells), and (N) TCF1+PD-1

Article Snippet: In vivo antibody therapies Rat-a-mouse CD8a (BioXCell, Lebanon, NH, clone: 53-6.7, #BE0004) or rat IgG2a isotype control (BioXcell, clone: 2A3, #BE0089) antibodies were administered i.v. every 6 days (200 mg/dose), starting at day -2 from tumor implantation for depleting CD8+ T cells.

Techniques: Construct, Tumor Implantation, Control, Cytometry, Expressing

Journal: Cancer Cell

Article Title: Genetic and pharmacological modulation of DNA mismatch repair heterogeneous tumors promotes immune surveillance

doi: 10.1016/j.ccell.2022.12.003

Figure Lengend Snippet: Immune profiling of MMR heterogeneous tumors reveals the involvement of CD8 + and γδ T cells (A) MMR heterogeneous tumors (100% Mlh1 +/+ , 80% Mlh1 +/+ 20% Mlh1 −/− , 50% Mlh1 +/+ 50% Mlh1 −/− ) were analyzed 13 days after injection in immunocompetent BALB C mice. Immune cell infiltrate was evaluated by flow cytometry. Percentages of CD8 + T, CD4 + T, γδ T (γδ-TCR + ), T reg (CD4 + FoxP3 + CD25 + ), B (CD19 + ), natural killer cells (CD49b + ), and macrophages (F4/80 + ), were calculated normalizing the absolute number of each population with the viable CD45 + fraction. The total amount of CD45 + alive cells is reported. Data are represented as average % ± standard error of the mean. The experiment was performed once. Tumors with insufficient material for immunophenotypical characterization were excluded from the analyses; n = 10 for 100% Mlh1 +/+ group, n = 9 for 80% Mlh1 +/+ 20% Mlh1 −/− group, n = 6 for 50% Mlh1 +/+ 50% Mlh1 −/− group. Statistical significance for each mixed population compared to the 100% Mlh1 +/+ control group was calculated by one-way ANOVA: ∗ p < 0.05 ∗∗ p < 0.005 ∗∗∗ p < 0.0005; (B) Ex vivo reactivity assays; CT26 100% Mlh1 +/+ and 50% Mlh1 +/+ 50% Mlh1 −/− were injected in immunocompetent BALB C mice. On day 13, mice were sacrificed and blood was taken. PBMCs were isolated and cocultured with Mlh1 +/+ or Mlh1 −/− separately (ratio PBMCs to tumor cells of 1:1). PBMCs harvested from naive mice were used as controls. IFN-γ expression was analyzed by flow cytometry as marker of activation after 5 h of coculture. The analyses of IFN-γ + CD8 + T, IFN-γ + CD4 + T, IFN-γ + γδ T cells obtained from the coculture with Mlh1 +/+ cells or (C) Mlh1 −/− cells is reported. At least three biological replicates were performed for each of the immune cell populations. Each dot represents a single biological replicate. Only conditions in which at least two biological replicates were above the threshold of 1% of IFNγ + cells (after background subtraction) were considered specifically activated. Mean ± standard error of the mean is reported (D) CT26 50% Mlh1 −/− 50% Mlh1 +/+ cells were injected in immunocompetent BALB C mice treated with depleting antibody for CD8 + T cells or γδ T cells. Twelve mice per group were used. Untreated mice served as control. Depleting CD8 + T cell antibody was given according to the following schedule: 400 μg at the day of the injection, 200 μg at the day 2, and then every 3 days after injection. Depleting γδ T cell antibody was administered according to the following schedule: 400 μg 2 days and 1 day before the injection, 400 μg at days 3 and 6 after injection followed by 400 μg every 7 days. Tumors were measured twice a week and volumes are reported as average tumor volume (mm 3 ) ± standard error of the mean; Mann-Whitney statistical analyses was performed, ∗ ∗∗∗ p < 0.0001. (E) Single mice graphs of the experiment shown in Figure 5 D are reported. (F) gDNA was extracted from four immune escaped tumors per arm (randomly selected), and ddPCR analyses was performed to determine the Mlh1 +/+ Mlh1 −/− cell content. Two or three sampling for each tumor were analyzed. Day 0 indicates the day of tumor cell injection. Data are represented as average % of cells ± standard deviation for each tumor. The experiment was performed once. See also Figure S3 .

Article Snippet: In Vivo Mab Anti-Mouse CD8a , Bioxcell , Cat#BE0061; RRID: AB_1125541.

Techniques: Injection, Flow Cytometry, Control, Ex Vivo, Isolation, Expressing, Marker, Activation Assay, MANN-WHITNEY, Sampling, Standard Deviation

Journal: Cancer Cell

Article Title: Genetic and pharmacological modulation of DNA mismatch repair heterogeneous tumors promotes immune surveillance

doi: 10.1016/j.ccell.2022.12.003

Figure Lengend Snippet:

Article Snippet: In Vivo Mab Anti-Mouse CD8a , Bioxcell , Cat#BE0061; RRID: AB_1125541.

Techniques: In Vivo, Recombinant, Purification, Virus, Staining, Sequencing, Detection Assay, Mutagenesis, Software

Journal: Cancers

Article Title: Antigens Expressed by Breast Cancer Cells Undergoing EMT Stimulate Cytotoxic CD8 + T Cell Immunity

doi: 10.3390/cancers14184397

Figure Lengend Snippet: CD8 + T cells are required for protection against mesenchymal-like EO771 tumor growth using EO771 whole cell vaccine (WCV). ( A ) Overview of tumor studies is shown. ( B ) Tumor growth curve of EO771 cells in mice treated with adjuvant alone ( n = 10) or EO771 WCV ( n = 10). Error bars depict mean with SEM. A two-way ANOVA was performed, and significant protection was provided by the EO771 WCV ( p = 0.0052). This experiment represents 4 tumor studies. ( C ) Tumor growth curve of EO771 cells in mice treated with adjuvant ( n = 16), EO771 WCV ( n = 16) or EO771 WCV plus 53-6.7 mAb ( n = 16). Graph represents 2 independent experiments; error bars depict mean with SEM. A two-way ANOVA was performed using Dunnett’s multiple comparisons test. At day 50, there was significant protection against tumor growth in mice treated with EO771 WCV ( p < 0.0001), which was partly reversed in mice that were also CD8-deficient ( p = 0.0157). ( D ) Representative plots from flow cytometric analysis depicting CD8 + T cell depletion from the peripheral blood of mice treated with CD8 + T cell depleting mAb (clone 53-6.7) or isotype control mAb (clone 2A3).

Article Snippet: Three days prior to primary vaccination, mice were treated with 500 μg of In Vivo Plus anti-mouse CD8a antibody (53-6.7, BioXCell, Lebanon, NH, USA, Cat. #BP0004-1) injected IP (100 μL in PBS) followed by weekly injections of 250 μg.

Techniques: Adjuvant, Control

Journal: Cancers

Article Title: Antigens Expressed by Breast Cancer Cells Undergoing EMT Stimulate Cytotoxic CD8 + T Cell Immunity

doi: 10.3390/cancers14184397

Figure Lengend Snippet: Restoration of miR-200c in EO771 alters expression of MHC class I SNV-derived neoantigens and putative SNV-derived peptides upregulated in mesenchymal-like EO771 cells elicit superior cytotoxic responses against epithelial-like EO771-miR-200c+ cells. ( A ) Overview of study design vaccinating mice with pooled MHC class I peptides followed by ex vivo T cell assays. ( B ) Quantification of putative peptides resulting from somatic variants per H-2D b /H-2K b allele with strong binding affinity predictions (<500 nM) is shown. The gray bar fraction corresponds to the number of peptides with significant change in expression ± miR-200c. ( C ) Flow cytometry analysis depicting CD8 + T cell proliferation in response to stimulation with individual MHC class I SNV-derived peptides. The gray peaks represent the dilution of cell trace violet (CTV) stain as CD8 + T cells proliferate in response to specific peptide. The black peak represents CTV stain in non-proliferating CD8 + T cells treated with negative control (no peptide). ( D ) Quantification of percent cytotoxicity and ( E ) IFNγ measurements from splenocytes of mice vaccinated with pooled peptides expressed more in the mesenchymal conditions and challenged with EO771-B7.1 ± miR-200c target cells ex vivo at the respective Effector: Target cell ratio (E:T). A two-way ANOVA was performed using Sidak’s multiple comparisons and found significant differences in cytotoxicity against EO771-B7.1-miR-200c target cells at both E:T ratios ( p < 0.0001). The numerical p values listed in the figure legend correlate with the asterisks in the figure.

Article Snippet: Three days prior to primary vaccination, mice were treated with 500 μg of In Vivo Plus anti-mouse CD8a antibody (53-6.7, BioXCell, Lebanon, NH, USA, Cat. #BP0004-1) injected IP (100 μL in PBS) followed by weekly injections of 250 μg.

Techniques: Expressing, Derivative Assay, Immunopeptidomics, Ex Vivo, Binding Assay, Flow Cytometry, Staining, Negative Control