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: Journal of Extracellular Vesicles

Article Title: A multiantigenic antibacterial nanovaccine utilizing hybrid membrane vesicles for combating Pseudomonas aeruginosa infections

doi: 10.1002/jev2.12524

Figure Lengend Snippet: In vivo immune responses activated by AuNP@HMV@SPs. (a) Schematic illustration of experimental design to evaluate the in vivo immune responses activated by AuNP@HMV@SPs (OMVs: 2 µg/mL, SP: 1.5 µg/mL) via subcutaneous injection. (b) Percentage of CD80 + and CD86 + cells (gated on CD11c + cells) in the lymph nodes collected on day 21 ( n = 4). (c) Percentage of GL7 + cells (gated on CD19 + IgG − cells) in the spleen collected on day 21 ( n = 5). (d) Anti‐ P. aeruginosa specific antibody IgG titers in serum collected on day 21 ( n = 5). (e) CD3 + T cells, (f) CD3 + CD4 + T cells, and (g) CD3 + CD8 + T cells in the spleen collected on day 21 ( n = 5). Naive mice without immunization were used as control. Statistical analysis was conducted using the student's t ‐test or One‐way ANOVA with Tukey's post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001 and ns representing non‐significance.

Article Snippet: FITC‐conjugated anti‐mouse CD11c, PE‐conjugated anti‐mouse CD80, APC‐conjugated anti‐mouse CD86, APC‐conjugated anti‐mouse CD3, PE‐conjugated anti‐mouse CD8a, FITC‐conjugated anti‐mouse CD4, and PE‐conjugated anti‐mouse CD19 were purchased from Tonbo Biosciences, U.S.A.. APC‐conjugated anti‐mouse GL‐7 was obtained from Laizee Biotech, Shanghai, China.

Techniques: In Vivo, Injection, Control

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: Cell reports

Article Title: Alpha synuclein, the culprit in Parkinson disease, is required for normal immune function

doi: 10.1016/j.celrep.2021.110090

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-mouse CD8a Ab, APC/Cy7 (clone 53-6.7) , Tonbo Biosciences , Cat# 25-0081-U100.

Techniques: Recombinant, Saline, Stripping Membranes, Reverse Transcription, SYBR Green Assay, RNAscope, Multiplex Assay, Polymer, Software, Flow Cytometry, Control, Light Microscopy