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cd8a depletion antibody 2.43  (Leinco Technologies)

 
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    Leinco Technologies cd8a depletion antibody 2.43
    VEEV-TC83 infection induces acute infiltration of T-cells in the olfactory bulb. A) Representative flow cytometry plots for identifying <t>CD8+</t> T cells from the brain (IC, 5dpi, TC83). B) Absolute counts of immune cells in brains, (n=4, IC, 5dpi, TC83). C) Quantification of flow cytometry analysis identifying total CD8+ T cells in the brains of mice over time, (n=4-6, IC, TC83). D-E) IHCp of OBs from mock or infected mice (7dpi, IN, TC83). D) Representative images of OBs stained with IHCp (Dapi=blue CD8=white Neun=pink). E) Quantification of CD8+ T cells per random 40x high powered field (hpf). (n=30 fields, N=6 OBs, * p ≤0.05).
    Cd8a Depletion Antibody 2.43, supplied by Leinco Technologies, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd8a+depletion+antibody+2%2E43/%CE%B1cd8+antibody/bio_rxiv__2025__05__13__653736-151-17-20
    Average 90 stars, based on 1 article reviews
    cd8a depletion antibody 2.43 - by Bioz Stars, 2026-09
    90/100 stars

    Images

    1) Product Images from "Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection"

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection

    Journal: bioRxiv

    doi: 10.1101/2025.05.13.653736

    VEEV-TC83 infection induces acute infiltration of T-cells in the olfactory bulb. A) Representative flow cytometry plots for identifying CD8+ T cells from the brain (IC, 5dpi, TC83). B) Absolute counts of immune cells in brains, (n=4, IC, 5dpi, TC83). C) Quantification of flow cytometry analysis identifying total CD8+ T cells in the brains of mice over time, (n=4-6, IC, TC83). D-E) IHCp of OBs from mock or infected mice (7dpi, IN, TC83). D) Representative images of OBs stained with IHCp (Dapi=blue CD8=white Neun=pink). E) Quantification of CD8+ T cells per random 40x high powered field (hpf). (n=30 fields, N=6 OBs, * p ≤0.05).
    Figure Legend Snippet: VEEV-TC83 infection induces acute infiltration of T-cells in the olfactory bulb. A) Representative flow cytometry plots for identifying CD8+ T cells from the brain (IC, 5dpi, TC83). B) Absolute counts of immune cells in brains, (n=4, IC, 5dpi, TC83). C) Quantification of flow cytometry analysis identifying total CD8+ T cells in the brains of mice over time, (n=4-6, IC, TC83). D-E) IHCp of OBs from mock or infected mice (7dpi, IN, TC83). D) Representative images of OBs stained with IHCp (Dapi=blue CD8=white Neun=pink). E) Quantification of CD8+ T cells per random 40x high powered field (hpf). (n=30 fields, N=6 OBs, * p ≤0.05).

    Techniques Used: Infection, Flow Cytometry, Staining

    VEEV-TC83 infection induces acute infiltration of bystander, memory CD8+ T-cells. A) Representative flow cytometry histograms comparing expression of bystander markers on CD8+ T cells between brains of mock and TC83-infected mice (n=4, 3dpi, IC). B) Representative flow cytometry histograms comparing expression of bystander markers on CD8+ T cells between 3dpi and 10dpi (n=4, IC, TC83). C) Representative flow contour plots showing CD44 and NKG2D expression on CD8+ T cells at 3dpi (upper panels) and 10dpi (lower panels) (n=4, 3 or 10dpi, IC, TC83) (Gating: Scatter, singlets, live, CD45+, CD2+CD3+, CD8+). D) Representative IHCp images from OBs of mock (upper) or infected (lower panel) IL15-reporter mice stained with Dapi=blue, CD45=cyan, TMEM119=green, CD8=white, CD3=orange, GFP-IL15=red, Neun=pink (7dpi, IN, TC83). e) qPCR of bystander-associated cytokines (n=4, 5dpi, IC),
    Figure Legend Snippet: VEEV-TC83 infection induces acute infiltration of bystander, memory CD8+ T-cells. A) Representative flow cytometry histograms comparing expression of bystander markers on CD8+ T cells between brains of mock and TC83-infected mice (n=4, 3dpi, IC). B) Representative flow cytometry histograms comparing expression of bystander markers on CD8+ T cells between 3dpi and 10dpi (n=4, IC, TC83). C) Representative flow contour plots showing CD44 and NKG2D expression on CD8+ T cells at 3dpi (upper panels) and 10dpi (lower panels) (n=4, 3 or 10dpi, IC, TC83) (Gating: Scatter, singlets, live, CD45+, CD2+CD3+, CD8+). D) Representative IHCp images from OBs of mock (upper) or infected (lower panel) IL15-reporter mice stained with Dapi=blue, CD45=cyan, TMEM119=green, CD8=white, CD3=orange, GFP-IL15=red, Neun=pink (7dpi, IN, TC83). e) qPCR of bystander-associated cytokines (n=4, 5dpi, IC),

    Techniques Used: Infection, Flow Cytometry, Expressing, Staining

    Microglia and macrophage-dependent IL15 production supports CD8+ T-cell recruitment and cytotoxicity. A) Upper panel: Representative flow plots of macrophage (CD11bhiCD45+, orange) and microglial (CD11b mid CD45+, pink) populations. Lower panel: histogram plots of IL15-GFP expression in macrophages (orange), microglia (pink) and T cells (black). (Gating: scatter, singlets, live; n=4, IC). B) Quantification of CD8+ T cells in the brains of infected WT or IL15ko mice (n=7, IC). C) Representative flow plots of cytokine and surface marker expression from CD8+ T cells from WT (upper panels) or IL15ko (lower panels) mice, all infected with TC83. (n=6, IC). D) Quantification of cytokine expression in CD8+ T cells of WT or IL15ko mice (IFNγ: n=4; Gzmb: n=3, all IC). E) Flow cytometric analysis of CD8+ T-cells from brains of WT and IL15KO mice indicating CD44 high NKG2D+ cells (n=6, IC). F) Mice were treated with vehicle or rIL15 (5ug) at day 0 and again 48hrs later. Mice were harvested 48hrs post first dose (n=4, IC). G) Quantification of protein expression from flow cytometric analysis (top panels) of CD8+ T cells with representative flow histograms (lower panels) from vehicle or rIL15 (5ug) treated mice (vehicle=light blue, rIL15=dark blue; IC, n=4). H) Representative flow plots of CD44 high NKG2D+ CD8+ T-cells in vehicle or rIL15 treated mice (n=4, IC).
    Figure Legend Snippet: Microglia and macrophage-dependent IL15 production supports CD8+ T-cell recruitment and cytotoxicity. A) Upper panel: Representative flow plots of macrophage (CD11bhiCD45+, orange) and microglial (CD11b mid CD45+, pink) populations. Lower panel: histogram plots of IL15-GFP expression in macrophages (orange), microglia (pink) and T cells (black). (Gating: scatter, singlets, live; n=4, IC). B) Quantification of CD8+ T cells in the brains of infected WT or IL15ko mice (n=7, IC). C) Representative flow plots of cytokine and surface marker expression from CD8+ T cells from WT (upper panels) or IL15ko (lower panels) mice, all infected with TC83. (n=6, IC). D) Quantification of cytokine expression in CD8+ T cells of WT or IL15ko mice (IFNγ: n=4; Gzmb: n=3, all IC). E) Flow cytometric analysis of CD8+ T-cells from brains of WT and IL15KO mice indicating CD44 high NKG2D+ cells (n=6, IC). F) Mice were treated with vehicle or rIL15 (5ug) at day 0 and again 48hrs later. Mice were harvested 48hrs post first dose (n=4, IC). G) Quantification of protein expression from flow cytometric analysis (top panels) of CD8+ T cells with representative flow histograms (lower panels) from vehicle or rIL15 (5ug) treated mice (vehicle=light blue, rIL15=dark blue; IC, n=4). H) Representative flow plots of CD44 high NKG2D+ CD8+ T-cells in vehicle or rIL15 treated mice (n=4, IC).

    Techniques Used: Expressing, Infection, Marker

    Virus-induced cytotoxic bystander CD8+ T-cells in the brain are activated independent of the T-cell receptor. A) Quantification of flow cytometric analysis of CD8+ T cell numbers from mock or infected WT or OT1 TCR transgenic mice (n=3-4, IC, TC83). B) Representative flow plots of GP33 tetramer+CD8+ T cells from mock (upper panels) or TC83 infected (lower panels) WT or P14 TCR transgenic mice (n=5, IC). C) Left panel: Quantification of total CD8+ T cell numbers per brain. Middle panel: Quantification of GP33-specific CD8+ T-cells from B. Right panel: Quantification of Tbet+ CD8+ T-cells per brain. (IC, TC83, n=4-5, p ≤0.5, one-way ANOVA with Multiple Comparisons post analysis). D) Representative flow plots of CD44 high NKG2D+ CD8+ T cells from P14 TCR transgenic mice mock (upper panel) or TC83 infected (lower panel) (n=4-5, i.c., TC83).
    Figure Legend Snippet: Virus-induced cytotoxic bystander CD8+ T-cells in the brain are activated independent of the T-cell receptor. A) Quantification of flow cytometric analysis of CD8+ T cell numbers from mock or infected WT or OT1 TCR transgenic mice (n=3-4, IC, TC83). B) Representative flow plots of GP33 tetramer+CD8+ T cells from mock (upper panels) or TC83 infected (lower panels) WT or P14 TCR transgenic mice (n=5, IC). C) Left panel: Quantification of total CD8+ T cell numbers per brain. Middle panel: Quantification of GP33-specific CD8+ T-cells from B. Right panel: Quantification of Tbet+ CD8+ T-cells per brain. (IC, TC83, n=4-5, p ≤0.5, one-way ANOVA with Multiple Comparisons post analysis). D) Representative flow plots of CD44 high NKG2D+ CD8+ T cells from P14 TCR transgenic mice mock (upper panel) or TC83 infected (lower panel) (n=4-5, i.c., TC83).

    Techniques Used: Virus, Infection, Transgenic Assay

    Bystander CD8+ T cells from the brains of TC83-infected mice exhibit cytotoxic characteristics. Left panels are quantification, right panels are representative flow plots. A) Flow cytometric analysis of CD107a expression in CD8+ T cells (n=5, IC, TC83). B) Flow cytometric analysis of IFNγ production in CD8+ T cells (n=6, IC, TC83). C) Flow cytometric analysis of Gzmb expression in CD8+ T cells (n=5, IC, TC83). * p ≤0.05, Student’s t-test.
    Figure Legend Snippet: Bystander CD8+ T cells from the brains of TC83-infected mice exhibit cytotoxic characteristics. Left panels are quantification, right panels are representative flow plots. A) Flow cytometric analysis of CD107a expression in CD8+ T cells (n=5, IC, TC83). B) Flow cytometric analysis of IFNγ production in CD8+ T cells (n=6, IC, TC83). C) Flow cytometric analysis of Gzmb expression in CD8+ T cells (n=5, IC, TC83). * p ≤0.05, Student’s t-test.

    Techniques Used: Infection, Expressing

    Bystander CD8+ T cell infiltration and activation is dependent on type 1 interferon. Mice were treated IC with vehicle (PBS) or PolyI:C, IFNa, or IFNb at 0hrs and again at 48hrs. Mice were harvested 4 days after first injection. A) Gene expression from mice treated with vehicle or PolyI:C (n=4). B) Representative flow plots of CD44hiNKG2D+ CD8+ T cells from PBS or PolyI:C treated mice. C) Gene expression from IFNα or IFNβ treated mice (n=4). D) Representative flow plots of CD44hiNKG2D+ CD8+ T cells from PBS, IFNα, or IFNβ treated mice. E) Quantification of CD8+ T-cells (left panel) and Tbet+CD8+ T cells (right panel) per brain from PBS, IFNα, or IFNβ treated mice. F) Representative flow cytometry histograms of activation markers on CD8+ T-cells from PBS (black outline), IFNα (blue), or IFNβ (green) treated mice (n=4). ** p≤ 0.05, student’s t-test or one-way ANOVA with multiple comparisons post analysis).
    Figure Legend Snippet: Bystander CD8+ T cell infiltration and activation is dependent on type 1 interferon. Mice were treated IC with vehicle (PBS) or PolyI:C, IFNa, or IFNb at 0hrs and again at 48hrs. Mice were harvested 4 days after first injection. A) Gene expression from mice treated with vehicle or PolyI:C (n=4). B) Representative flow plots of CD44hiNKG2D+ CD8+ T cells from PBS or PolyI:C treated mice. C) Gene expression from IFNα or IFNβ treated mice (n=4). D) Representative flow plots of CD44hiNKG2D+ CD8+ T cells from PBS, IFNα, or IFNβ treated mice. E) Quantification of CD8+ T-cells (left panel) and Tbet+CD8+ T cells (right panel) per brain from PBS, IFNα, or IFNβ treated mice. F) Representative flow cytometry histograms of activation markers on CD8+ T-cells from PBS (black outline), IFNα (blue), or IFNβ (green) treated mice (n=4). ** p≤ 0.05, student’s t-test or one-way ANOVA with multiple comparisons post analysis).

    Techniques Used: Activation Assay, Injection, Gene Expression, Flow Cytometry

    Bystander CD8+ T cells contribute to CNS injury following virus infection. A) Rag-/-mice and WT mice were i.c. infected with TC83 and viral genome copies in the brain were measured with qPCR (5dpi, n=8). B) At the time of ic infection, WT mice were injected ip with either isotype control antibody or CD8-depletion antibody. Viral genome copies were measured with qPCR (5dpi, TC83, n=8). C) Mice were infected IN with TC83 and either injected IP with isotype control antibody or CD8-depletion antibody. At 5dpi, brains were harvested and processed for IHCp. Representative images of the OBs of infected mice (Dapi=blue, Neu=pink, cl-caspase3=white). D) Cleaved caspase 3 was quantified with IHCp of individual OBs in neurons (Neun+), astroglia (GFAP+), and microglia (Iba1+) (n=4, p ≤0.05 Student’s t test). E) Phosphorylated Stat1 was quantified with IHCp of individual OBs in neurons (Neun+), astrocytes (GFAP+), and microglia (Iba1+) (n=4, * p ≤0.05 Student’s t test).
    Figure Legend Snippet: Bystander CD8+ T cells contribute to CNS injury following virus infection. A) Rag-/-mice and WT mice were i.c. infected with TC83 and viral genome copies in the brain were measured with qPCR (5dpi, n=8). B) At the time of ic infection, WT mice were injected ip with either isotype control antibody or CD8-depletion antibody. Viral genome copies were measured with qPCR (5dpi, TC83, n=8). C) Mice were infected IN with TC83 and either injected IP with isotype control antibody or CD8-depletion antibody. At 5dpi, brains were harvested and processed for IHCp. Representative images of the OBs of infected mice (Dapi=blue, Neu=pink, cl-caspase3=white). D) Cleaved caspase 3 was quantified with IHCp of individual OBs in neurons (Neun+), astroglia (GFAP+), and microglia (Iba1+) (n=4, p ≤0.05 Student’s t test). E) Phosphorylated Stat1 was quantified with IHCp of individual OBs in neurons (Neun+), astrocytes (GFAP+), and microglia (Iba1+) (n=4, * p ≤0.05 Student’s t test).

    Techniques Used: Virus, Infection, Injection, Control

    Related Articles

    Injection:

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection
    Article Snippet: LCMV Armstrong stocks were propagated in baby hamster kidney 21 cells and generously provided by Dr. Raymond M Welsh and amplified in the Berg laboratory. .. To deplete CD8+ T-cells, mice were injected i.p. with 250ug of isotype control (Leinco, clone GL113) or CD8a depletion antibody (Leinco, clone 2.43) at the time of infection with TC83. ..

    Control:

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection
    Article Snippet: LCMV Armstrong stocks were propagated in baby hamster kidney 21 cells and generously provided by Dr. Raymond M Welsh and amplified in the Berg laboratory. .. To deplete CD8+ T-cells, mice were injected i.p. with 250ug of isotype control (Leinco, clone GL113) or CD8a depletion antibody (Leinco, clone 2.43) at the time of infection with TC83. ..

    Infection:

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection
    Article Snippet: LCMV Armstrong stocks were propagated in baby hamster kidney 21 cells and generously provided by Dr. Raymond M Welsh and amplified in the Berg laboratory. .. To deplete CD8+ T-cells, mice were injected i.p. with 250ug of isotype control (Leinco, clone GL113) or CD8a depletion antibody (Leinco, clone 2.43) at the time of infection with TC83. ..



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    Image Search Results


    VEEV-TC83 infection induces acute infiltration of T-cells in the olfactory bulb. A) Representative flow cytometry plots for identifying CD8+ T cells from the brain (IC, 5dpi, TC83). B) Absolute counts of immune cells in brains, (n=4, IC, 5dpi, TC83). C) Quantification of flow cytometry analysis identifying total CD8+ T cells in the brains of mice over time, (n=4-6, IC, TC83). D-E) IHCp of OBs from mock or infected mice (7dpi, IN, TC83). D) Representative images of OBs stained with IHCp (Dapi=blue CD8=white Neun=pink). E) Quantification of CD8+ T cells per random 40x high powered field (hpf). (n=30 fields, N=6 OBs, * p ≤0.05).

    Journal: bioRxiv

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection

    doi: 10.1101/2025.05.13.653736

    Figure Lengend Snippet: VEEV-TC83 infection induces acute infiltration of T-cells in the olfactory bulb. A) Representative flow cytometry plots for identifying CD8+ T cells from the brain (IC, 5dpi, TC83). B) Absolute counts of immune cells in brains, (n=4, IC, 5dpi, TC83). C) Quantification of flow cytometry analysis identifying total CD8+ T cells in the brains of mice over time, (n=4-6, IC, TC83). D-E) IHCp of OBs from mock or infected mice (7dpi, IN, TC83). D) Representative images of OBs stained with IHCp (Dapi=blue CD8=white Neun=pink). E) Quantification of CD8+ T cells per random 40x high powered field (hpf). (n=30 fields, N=6 OBs, * p ≤0.05).

    Article Snippet: To deplete CD8+ T-cells, mice were injected i.p. with 250ug of isotype control (Leinco, clone GL113) or CD8a depletion antibody (Leinco, clone 2.43) at the time of infection with TC83.

    Techniques: Infection, Flow Cytometry, Staining

    VEEV-TC83 infection induces acute infiltration of bystander, memory CD8+ T-cells. A) Representative flow cytometry histograms comparing expression of bystander markers on CD8+ T cells between brains of mock and TC83-infected mice (n=4, 3dpi, IC). B) Representative flow cytometry histograms comparing expression of bystander markers on CD8+ T cells between 3dpi and 10dpi (n=4, IC, TC83). C) Representative flow contour plots showing CD44 and NKG2D expression on CD8+ T cells at 3dpi (upper panels) and 10dpi (lower panels) (n=4, 3 or 10dpi, IC, TC83) (Gating: Scatter, singlets, live, CD45+, CD2+CD3+, CD8+). D) Representative IHCp images from OBs of mock (upper) or infected (lower panel) IL15-reporter mice stained with Dapi=blue, CD45=cyan, TMEM119=green, CD8=white, CD3=orange, GFP-IL15=red, Neun=pink (7dpi, IN, TC83). e) qPCR of bystander-associated cytokines (n=4, 5dpi, IC),

    Journal: bioRxiv

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection

    doi: 10.1101/2025.05.13.653736

    Figure Lengend Snippet: VEEV-TC83 infection induces acute infiltration of bystander, memory CD8+ T-cells. A) Representative flow cytometry histograms comparing expression of bystander markers on CD8+ T cells between brains of mock and TC83-infected mice (n=4, 3dpi, IC). B) Representative flow cytometry histograms comparing expression of bystander markers on CD8+ T cells between 3dpi and 10dpi (n=4, IC, TC83). C) Representative flow contour plots showing CD44 and NKG2D expression on CD8+ T cells at 3dpi (upper panels) and 10dpi (lower panels) (n=4, 3 or 10dpi, IC, TC83) (Gating: Scatter, singlets, live, CD45+, CD2+CD3+, CD8+). D) Representative IHCp images from OBs of mock (upper) or infected (lower panel) IL15-reporter mice stained with Dapi=blue, CD45=cyan, TMEM119=green, CD8=white, CD3=orange, GFP-IL15=red, Neun=pink (7dpi, IN, TC83). e) qPCR of bystander-associated cytokines (n=4, 5dpi, IC),

    Article Snippet: To deplete CD8+ T-cells, mice were injected i.p. with 250ug of isotype control (Leinco, clone GL113) or CD8a depletion antibody (Leinco, clone 2.43) at the time of infection with TC83.

    Techniques: Infection, Flow Cytometry, Expressing, Staining

    Microglia and macrophage-dependent IL15 production supports CD8+ T-cell recruitment and cytotoxicity. A) Upper panel: Representative flow plots of macrophage (CD11bhiCD45+, orange) and microglial (CD11b mid CD45+, pink) populations. Lower panel: histogram plots of IL15-GFP expression in macrophages (orange), microglia (pink) and T cells (black). (Gating: scatter, singlets, live; n=4, IC). B) Quantification of CD8+ T cells in the brains of infected WT or IL15ko mice (n=7, IC). C) Representative flow plots of cytokine and surface marker expression from CD8+ T cells from WT (upper panels) or IL15ko (lower panels) mice, all infected with TC83. (n=6, IC). D) Quantification of cytokine expression in CD8+ T cells of WT or IL15ko mice (IFNγ: n=4; Gzmb: n=3, all IC). E) Flow cytometric analysis of CD8+ T-cells from brains of WT and IL15KO mice indicating CD44 high NKG2D+ cells (n=6, IC). F) Mice were treated with vehicle or rIL15 (5ug) at day 0 and again 48hrs later. Mice were harvested 48hrs post first dose (n=4, IC). G) Quantification of protein expression from flow cytometric analysis (top panels) of CD8+ T cells with representative flow histograms (lower panels) from vehicle or rIL15 (5ug) treated mice (vehicle=light blue, rIL15=dark blue; IC, n=4). H) Representative flow plots of CD44 high NKG2D+ CD8+ T-cells in vehicle or rIL15 treated mice (n=4, IC).

    Journal: bioRxiv

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection

    doi: 10.1101/2025.05.13.653736

    Figure Lengend Snippet: Microglia and macrophage-dependent IL15 production supports CD8+ T-cell recruitment and cytotoxicity. A) Upper panel: Representative flow plots of macrophage (CD11bhiCD45+, orange) and microglial (CD11b mid CD45+, pink) populations. Lower panel: histogram plots of IL15-GFP expression in macrophages (orange), microglia (pink) and T cells (black). (Gating: scatter, singlets, live; n=4, IC). B) Quantification of CD8+ T cells in the brains of infected WT or IL15ko mice (n=7, IC). C) Representative flow plots of cytokine and surface marker expression from CD8+ T cells from WT (upper panels) or IL15ko (lower panels) mice, all infected with TC83. (n=6, IC). D) Quantification of cytokine expression in CD8+ T cells of WT or IL15ko mice (IFNγ: n=4; Gzmb: n=3, all IC). E) Flow cytometric analysis of CD8+ T-cells from brains of WT and IL15KO mice indicating CD44 high NKG2D+ cells (n=6, IC). F) Mice were treated with vehicle or rIL15 (5ug) at day 0 and again 48hrs later. Mice were harvested 48hrs post first dose (n=4, IC). G) Quantification of protein expression from flow cytometric analysis (top panels) of CD8+ T cells with representative flow histograms (lower panels) from vehicle or rIL15 (5ug) treated mice (vehicle=light blue, rIL15=dark blue; IC, n=4). H) Representative flow plots of CD44 high NKG2D+ CD8+ T-cells in vehicle or rIL15 treated mice (n=4, IC).

    Article Snippet: To deplete CD8+ T-cells, mice were injected i.p. with 250ug of isotype control (Leinco, clone GL113) or CD8a depletion antibody (Leinco, clone 2.43) at the time of infection with TC83.

    Techniques: Expressing, Infection, Marker

    Virus-induced cytotoxic bystander CD8+ T-cells in the brain are activated independent of the T-cell receptor. A) Quantification of flow cytometric analysis of CD8+ T cell numbers from mock or infected WT or OT1 TCR transgenic mice (n=3-4, IC, TC83). B) Representative flow plots of GP33 tetramer+CD8+ T cells from mock (upper panels) or TC83 infected (lower panels) WT or P14 TCR transgenic mice (n=5, IC). C) Left panel: Quantification of total CD8+ T cell numbers per brain. Middle panel: Quantification of GP33-specific CD8+ T-cells from B. Right panel: Quantification of Tbet+ CD8+ T-cells per brain. (IC, TC83, n=4-5, p ≤0.5, one-way ANOVA with Multiple Comparisons post analysis). D) Representative flow plots of CD44 high NKG2D+ CD8+ T cells from P14 TCR transgenic mice mock (upper panel) or TC83 infected (lower panel) (n=4-5, i.c., TC83).

    Journal: bioRxiv

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection

    doi: 10.1101/2025.05.13.653736

    Figure Lengend Snippet: Virus-induced cytotoxic bystander CD8+ T-cells in the brain are activated independent of the T-cell receptor. A) Quantification of flow cytometric analysis of CD8+ T cell numbers from mock or infected WT or OT1 TCR transgenic mice (n=3-4, IC, TC83). B) Representative flow plots of GP33 tetramer+CD8+ T cells from mock (upper panels) or TC83 infected (lower panels) WT or P14 TCR transgenic mice (n=5, IC). C) Left panel: Quantification of total CD8+ T cell numbers per brain. Middle panel: Quantification of GP33-specific CD8+ T-cells from B. Right panel: Quantification of Tbet+ CD8+ T-cells per brain. (IC, TC83, n=4-5, p ≤0.5, one-way ANOVA with Multiple Comparisons post analysis). D) Representative flow plots of CD44 high NKG2D+ CD8+ T cells from P14 TCR transgenic mice mock (upper panel) or TC83 infected (lower panel) (n=4-5, i.c., TC83).

    Article Snippet: To deplete CD8+ T-cells, mice were injected i.p. with 250ug of isotype control (Leinco, clone GL113) or CD8a depletion antibody (Leinco, clone 2.43) at the time of infection with TC83.

    Techniques: Virus, Infection, Transgenic Assay

    Bystander CD8+ T cells from the brains of TC83-infected mice exhibit cytotoxic characteristics. Left panels are quantification, right panels are representative flow plots. A) Flow cytometric analysis of CD107a expression in CD8+ T cells (n=5, IC, TC83). B) Flow cytometric analysis of IFNγ production in CD8+ T cells (n=6, IC, TC83). C) Flow cytometric analysis of Gzmb expression in CD8+ T cells (n=5, IC, TC83). * p ≤0.05, Student’s t-test.

    Journal: bioRxiv

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection

    doi: 10.1101/2025.05.13.653736

    Figure Lengend Snippet: Bystander CD8+ T cells from the brains of TC83-infected mice exhibit cytotoxic characteristics. Left panels are quantification, right panels are representative flow plots. A) Flow cytometric analysis of CD107a expression in CD8+ T cells (n=5, IC, TC83). B) Flow cytometric analysis of IFNγ production in CD8+ T cells (n=6, IC, TC83). C) Flow cytometric analysis of Gzmb expression in CD8+ T cells (n=5, IC, TC83). * p ≤0.05, Student’s t-test.

    Article Snippet: To deplete CD8+ T-cells, mice were injected i.p. with 250ug of isotype control (Leinco, clone GL113) or CD8a depletion antibody (Leinco, clone 2.43) at the time of infection with TC83.

    Techniques: Infection, Expressing

    Bystander CD8+ T cell infiltration and activation is dependent on type 1 interferon. Mice were treated IC with vehicle (PBS) or PolyI:C, IFNa, or IFNb at 0hrs and again at 48hrs. Mice were harvested 4 days after first injection. A) Gene expression from mice treated with vehicle or PolyI:C (n=4). B) Representative flow plots of CD44hiNKG2D+ CD8+ T cells from PBS or PolyI:C treated mice. C) Gene expression from IFNα or IFNβ treated mice (n=4). D) Representative flow plots of CD44hiNKG2D+ CD8+ T cells from PBS, IFNα, or IFNβ treated mice. E) Quantification of CD8+ T-cells (left panel) and Tbet+CD8+ T cells (right panel) per brain from PBS, IFNα, or IFNβ treated mice. F) Representative flow cytometry histograms of activation markers on CD8+ T-cells from PBS (black outline), IFNα (blue), or IFNβ (green) treated mice (n=4). ** p≤ 0.05, student’s t-test or one-way ANOVA with multiple comparisons post analysis).

    Journal: bioRxiv

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection

    doi: 10.1101/2025.05.13.653736

    Figure Lengend Snippet: Bystander CD8+ T cell infiltration and activation is dependent on type 1 interferon. Mice were treated IC with vehicle (PBS) or PolyI:C, IFNa, or IFNb at 0hrs and again at 48hrs. Mice were harvested 4 days after first injection. A) Gene expression from mice treated with vehicle or PolyI:C (n=4). B) Representative flow plots of CD44hiNKG2D+ CD8+ T cells from PBS or PolyI:C treated mice. C) Gene expression from IFNα or IFNβ treated mice (n=4). D) Representative flow plots of CD44hiNKG2D+ CD8+ T cells from PBS, IFNα, or IFNβ treated mice. E) Quantification of CD8+ T-cells (left panel) and Tbet+CD8+ T cells (right panel) per brain from PBS, IFNα, or IFNβ treated mice. F) Representative flow cytometry histograms of activation markers on CD8+ T-cells from PBS (black outline), IFNα (blue), or IFNβ (green) treated mice (n=4). ** p≤ 0.05, student’s t-test or one-way ANOVA with multiple comparisons post analysis).

    Article Snippet: To deplete CD8+ T-cells, mice were injected i.p. with 250ug of isotype control (Leinco, clone GL113) or CD8a depletion antibody (Leinco, clone 2.43) at the time of infection with TC83.

    Techniques: Activation Assay, Injection, Gene Expression, Flow Cytometry

    Bystander CD8+ T cells contribute to CNS injury following virus infection. A) Rag-/-mice and WT mice were i.c. infected with TC83 and viral genome copies in the brain were measured with qPCR (5dpi, n=8). B) At the time of ic infection, WT mice were injected ip with either isotype control antibody or CD8-depletion antibody. Viral genome copies were measured with qPCR (5dpi, TC83, n=8). C) Mice were infected IN with TC83 and either injected IP with isotype control antibody or CD8-depletion antibody. At 5dpi, brains were harvested and processed for IHCp. Representative images of the OBs of infected mice (Dapi=blue, Neu=pink, cl-caspase3=white). D) Cleaved caspase 3 was quantified with IHCp of individual OBs in neurons (Neun+), astroglia (GFAP+), and microglia (Iba1+) (n=4, p ≤0.05 Student’s t test). E) Phosphorylated Stat1 was quantified with IHCp of individual OBs in neurons (Neun+), astrocytes (GFAP+), and microglia (Iba1+) (n=4, * p ≤0.05 Student’s t test).

    Journal: bioRxiv

    Article Title: Myeloid cell IL-15 production in the brain supports Bystander CD8+ T-Cell Neuropathic Immune Responses following Virus infection

    doi: 10.1101/2025.05.13.653736

    Figure Lengend Snippet: Bystander CD8+ T cells contribute to CNS injury following virus infection. A) Rag-/-mice and WT mice were i.c. infected with TC83 and viral genome copies in the brain were measured with qPCR (5dpi, n=8). B) At the time of ic infection, WT mice were injected ip with either isotype control antibody or CD8-depletion antibody. Viral genome copies were measured with qPCR (5dpi, TC83, n=8). C) Mice were infected IN with TC83 and either injected IP with isotype control antibody or CD8-depletion antibody. At 5dpi, brains were harvested and processed for IHCp. Representative images of the OBs of infected mice (Dapi=blue, Neu=pink, cl-caspase3=white). D) Cleaved caspase 3 was quantified with IHCp of individual OBs in neurons (Neun+), astroglia (GFAP+), and microglia (Iba1+) (n=4, p ≤0.05 Student’s t test). E) Phosphorylated Stat1 was quantified with IHCp of individual OBs in neurons (Neun+), astrocytes (GFAP+), and microglia (Iba1+) (n=4, * p ≤0.05 Student’s t test).

    Article Snippet: To deplete CD8+ T-cells, mice were injected i.p. with 250ug of isotype control (Leinco, clone GL113) or CD8a depletion antibody (Leinco, clone 2.43) at the time of infection with TC83.

    Techniques: Virus, Infection, Injection, Control

    AlloDCs enhanced the therapeutic efficacy of α4-1BB antibody that dependent on CD8 + T-cells. (A) Schematic illustration of the experiment outline of the CT-26 model. (B) Mouse survival (Kaplan-Meier curve) in the CT-26 model after different treatments as indicated (PBS n=10, AlloDC n=14, α4-1BB n=15, α4-1BB/AlloDC n=10, α4-1BB/AlloDC/CD4 depletion n=6, α4-1BB/AlloDC/CD8 depletion n=6). (C, D) Tumor size in individual mouse and survival after re-challenge with CT-26 tumor. All mice that survived in (B) were re-challenged and none of them got tumor after re-challenge. The number of rechallenged mice from each group are labeled in figure panel. (E–J) Tumor size in individual mouse in different treatment groups as in (B) . (K) Schematic illustration of the experiment outline of the B16 model. (L) Mouse survival (Kaplan-Meier curve) in the B16 model after different treatments as indicated. (PBS n=10, AlloDC n=9, α4-1BB n=8, α4-1BB/AlloDC n=9). (M–P) Tumor size in individual mouse in different treatment groups as in (L) . The size of each group (n) is labeled in each panel below. All survival curves were compared using the Log-Rank test. (*P<0.05, **P<0.01, ****P<0.0001).

    Journal: Frontiers in Immunology

    Article Title: Proinflammatory allogeneic dendritic cells enhance the therapeutic efficacy of systemic anti-4-1BB treatment

    doi: 10.3389/fimmu.2023.1146413

    Figure Lengend Snippet: AlloDCs enhanced the therapeutic efficacy of α4-1BB antibody that dependent on CD8 + T-cells. (A) Schematic illustration of the experiment outline of the CT-26 model. (B) Mouse survival (Kaplan-Meier curve) in the CT-26 model after different treatments as indicated (PBS n=10, AlloDC n=14, α4-1BB n=15, α4-1BB/AlloDC n=10, α4-1BB/AlloDC/CD4 depletion n=6, α4-1BB/AlloDC/CD8 depletion n=6). (C, D) Tumor size in individual mouse and survival after re-challenge with CT-26 tumor. All mice that survived in (B) were re-challenged and none of them got tumor after re-challenge. The number of rechallenged mice from each group are labeled in figure panel. (E–J) Tumor size in individual mouse in different treatment groups as in (B) . (K) Schematic illustration of the experiment outline of the B16 model. (L) Mouse survival (Kaplan-Meier curve) in the B16 model after different treatments as indicated. (PBS n=10, AlloDC n=9, α4-1BB n=8, α4-1BB/AlloDC n=9). (M–P) Tumor size in individual mouse in different treatment groups as in (L) . The size of each group (n) is labeled in each panel below. All survival curves were compared using the Log-Rank test. (*P<0.05, **P<0.01, ****P<0.0001).

    Article Snippet: CD4 depletion antibody (In Vivo Mab αmouse CD4, clone GK1.5, BioCell) and CD8 depletion antibody (In Vivo Mab αmouse CD8a, clone 2.43, BioCell) were injected (i.p) on day 13, 14, 15, 20, 21 and 22.

    Techniques: Drug discovery, Labeling

    AlloDCs boost the lymphoid compartment signature for α4-1BB/AlloDC therapies. (A) The experimental setup is depicted in a schematic illustration. (B) The comparison between the PBS-treated group and the grouped samples based on up- or down-regulation is presented as the sum of pathway signature scores determined from NanoString mRNA profiling. (C) The radar map displays the pathway scores from different treatment groups. (D, E) The heatmap showcases the top 15 differentially expressed genes from lymphoid compartment pathways and cell proliferation. (F) The abundance of tumor-infiltrating immune cells in each mouse from different treatment groups is presented as cell-type scores from NanoString mRNA profiling. Error bars represent SEM and the mean values were compared using one-way ANOVA nonparametric test. (*P<0.05, **P<0.01).

    Journal: Frontiers in Immunology

    Article Title: Proinflammatory allogeneic dendritic cells enhance the therapeutic efficacy of systemic anti-4-1BB treatment

    doi: 10.3389/fimmu.2023.1146413

    Figure Lengend Snippet: AlloDCs boost the lymphoid compartment signature for α4-1BB/AlloDC therapies. (A) The experimental setup is depicted in a schematic illustration. (B) The comparison between the PBS-treated group and the grouped samples based on up- or down-regulation is presented as the sum of pathway signature scores determined from NanoString mRNA profiling. (C) The radar map displays the pathway scores from different treatment groups. (D, E) The heatmap showcases the top 15 differentially expressed genes from lymphoid compartment pathways and cell proliferation. (F) The abundance of tumor-infiltrating immune cells in each mouse from different treatment groups is presented as cell-type scores from NanoString mRNA profiling. Error bars represent SEM and the mean values were compared using one-way ANOVA nonparametric test. (*P<0.05, **P<0.01).

    Article Snippet: CD4 depletion antibody (In Vivo Mab αmouse CD4, clone GK1.5, BioCell) and CD8 depletion antibody (In Vivo Mab αmouse CD8a, clone 2.43, BioCell) were injected (i.p) on day 13, 14, 15, 20, 21 and 22.

    Techniques: Comparison

    AlloDCs combined with α4-1BB treatment attract CD8 + T-cell populations with activated phenotype by enhanced DC activation. (A) The ratio of CD8+ and CD4+ T-cells in the tumor-infiltrating T-cell population (gated as CD3+ T-cells) is analyzed using flow cytometry. (B) The percentage of IFNγ + CD8 + T-cells in the tumor is measured. (C) The percentage of CD107a + CD8 + T-cells in the tumor is determined. (D–G) The percentage of phenotypically exhausted CD8+ T-cells in the tumor is assessed, represented as triple, double, and single positive for PD-1 + , Tim3 + and LAG3 + markers on CD8 + T cells. (H) Percentage of tumor infiltrating tissue-resident memory (T RM ) CD8 + T-cells (gated as CD49a + CD103 + T-cells out of CD69 + CD8 + T-cells) in each treatment group, analyzed by flow cytometry. (I) Representative density plot showing tumor infiltrating T RM in each treatment group. (J) Percentage of tumor-reactive CD8 + T-cells (CD39 + CD103 + out of CD8 + T-cells) in tumor samples from different treatment groups. (K) IFN-γ expression level in the supernatant of in vitro cultured splenocytes, harvested from each treatment group and re-stimulated with either gp70 peptides or non-relevant peptides. (L) Percentage of the tumor-infiltrating DCs (CD11C + F4/80 - out of CD11b + CD45 + cells) in each treatment group. (M, N) The expression of CD103a and CD8a on the tumor infiltrating DCs. (O, P) Percentage of the CCL5 + and IL12 + tumor-infiltrating DCs in each treatment group. Error bars represent SEM and the mean values were compared using one-way ANOVA nonparametric test. (*P<0.05, **P<0.01).

    Journal: Frontiers in Immunology

    Article Title: Proinflammatory allogeneic dendritic cells enhance the therapeutic efficacy of systemic anti-4-1BB treatment

    doi: 10.3389/fimmu.2023.1146413

    Figure Lengend Snippet: AlloDCs combined with α4-1BB treatment attract CD8 + T-cell populations with activated phenotype by enhanced DC activation. (A) The ratio of CD8+ and CD4+ T-cells in the tumor-infiltrating T-cell population (gated as CD3+ T-cells) is analyzed using flow cytometry. (B) The percentage of IFNγ + CD8 + T-cells in the tumor is measured. (C) The percentage of CD107a + CD8 + T-cells in the tumor is determined. (D–G) The percentage of phenotypically exhausted CD8+ T-cells in the tumor is assessed, represented as triple, double, and single positive for PD-1 + , Tim3 + and LAG3 + markers on CD8 + T cells. (H) Percentage of tumor infiltrating tissue-resident memory (T RM ) CD8 + T-cells (gated as CD49a + CD103 + T-cells out of CD69 + CD8 + T-cells) in each treatment group, analyzed by flow cytometry. (I) Representative density plot showing tumor infiltrating T RM in each treatment group. (J) Percentage of tumor-reactive CD8 + T-cells (CD39 + CD103 + out of CD8 + T-cells) in tumor samples from different treatment groups. (K) IFN-γ expression level in the supernatant of in vitro cultured splenocytes, harvested from each treatment group and re-stimulated with either gp70 peptides or non-relevant peptides. (L) Percentage of the tumor-infiltrating DCs (CD11C + F4/80 - out of CD11b + CD45 + cells) in each treatment group. (M, N) The expression of CD103a and CD8a on the tumor infiltrating DCs. (O, P) Percentage of the CCL5 + and IL12 + tumor-infiltrating DCs in each treatment group. Error bars represent SEM and the mean values were compared using one-way ANOVA nonparametric test. (*P<0.05, **P<0.01).

    Article Snippet: CD4 depletion antibody (In Vivo Mab αmouse CD4, clone GK1.5, BioCell) and CD8 depletion antibody (In Vivo Mab αmouse CD8a, clone 2.43, BioCell) were injected (i.p) on day 13, 14, 15, 20, 21 and 22.

    Techniques: Activation Assay, Flow Cytometry, Expressing, In Vitro, Cell Culture

    CDK7 inhibition stimulates antitumor immunity and sensitizes NSCLC to antiPD-1 therapy. a Photographs of tumors from the Lewis lung cancer model treated with the combination of THZ1 and antiPD-1 antibody ( n = 7). b Tumor growth curves of mice from the Lewis lung cancer model. The dashed line represents a single mouse data in each group and the solid line represents the mean value in different groups. Error bars represent ± SEM (* P < 0.05; ** P < 0.01). c Weights of tumors from mice in Lewis lung cancer model at the endpoint (* P < 0.05; *** P < 0.001). d Quantitation of PD-L1 level on tumor surfaces from the Lewis lung cancer model (*** P < 0.001). e Quantification of percentages of CD45 + cells in tumors from the Lewis lung cancer model treated with the combination of THZ1 and antiPD-1 antibody (*** P < 0.001). f Quantification of percentage of CD45 + CD8 + cells in tumor from the Lewis lung cancer model treated with the combination of THZ1 and antiPD-1 antibody ( n = 7) (** P < 0.01; *** P < 0.001). g The ratios of CD8 + T cell/CD45 + cell in tumors from the Lewis lung cancer model (*** P < 0.001). h IFN-γ in the tumors from the Lewis lung cancer model was measured (*** P < 0.001). The collected tumors were homogenized and detected by using Quantikine ELISA (R&D Systems). i C57BL/6 mice bearing Lewis tumor were treated with THZ1 or the combination of THZ1 and antiPD-1 with or without CD8 + T depletion by antibodies. The tumor burden of different groups was quantified by tumor weights at the endpoint (* P < 0.05; *** P < 0.001). j Kaplan-Meier survival analysis of patients with different risk scores by CDK7 protein level and tumor-infiltrating lymphocyte (TIL) scores in cohort II. High CDK7 protein level and low TIL scores were defined as two risk factors. Patients were stratified into three risk groups with different survival outcomes as follows: low-risk group without any risk factors = low CDK7 protein level and high TIL score ( n = 64); medium risk group with only one risk factor ( n = 126); high-risk group with two risk factors = high CDK7 protein level and low TIL score ( n = 32) (* P < 0.05). k Kaplan-Meier survival analysis of patients with different risk scores by CDK7 protein level, MYC protein level, and TILs score in cohort II. High CDK7 protein level, high MYC protein level and low TIL score were defined as three risk factors. Patients were stratified into three risk groups with different survival outcomes as follows: low-risk group without any risk factors ( n = 41); medium risk group with only one risk factor ( n = 97); high-risk group with at least two risk factors ( n = 84) (** P < 0.01). l Schematic illustration showing the CDK7-p38α-MYC axis dependent regulation of PD-L1 and the function of this signaling in NSCLC. The solid arrows represent strong processes; the dashed arrows represent very weak processes after CDK7 inhibition

    Journal: Journal of Hematology & Oncology

    Article Title: CDK7 inhibitor THZ1 enhances antiPD-1 therapy efficacy via the p38α/MYC/PD-L1 signaling in non-small cell lung cancer

    doi: 10.1186/s13045-020-00926-x

    Figure Lengend Snippet: CDK7 inhibition stimulates antitumor immunity and sensitizes NSCLC to antiPD-1 therapy. a Photographs of tumors from the Lewis lung cancer model treated with the combination of THZ1 and antiPD-1 antibody ( n = 7). b Tumor growth curves of mice from the Lewis lung cancer model. The dashed line represents a single mouse data in each group and the solid line represents the mean value in different groups. Error bars represent ± SEM (* P < 0.05; ** P < 0.01). c Weights of tumors from mice in Lewis lung cancer model at the endpoint (* P < 0.05; *** P < 0.001). d Quantitation of PD-L1 level on tumor surfaces from the Lewis lung cancer model (*** P < 0.001). e Quantification of percentages of CD45 + cells in tumors from the Lewis lung cancer model treated with the combination of THZ1 and antiPD-1 antibody (*** P < 0.001). f Quantification of percentage of CD45 + CD8 + cells in tumor from the Lewis lung cancer model treated with the combination of THZ1 and antiPD-1 antibody ( n = 7) (** P < 0.01; *** P < 0.001). g The ratios of CD8 + T cell/CD45 + cell in tumors from the Lewis lung cancer model (*** P < 0.001). h IFN-γ in the tumors from the Lewis lung cancer model was measured (*** P < 0.001). The collected tumors were homogenized and detected by using Quantikine ELISA (R&D Systems). i C57BL/6 mice bearing Lewis tumor were treated with THZ1 or the combination of THZ1 and antiPD-1 with or without CD8 + T depletion by antibodies. The tumor burden of different groups was quantified by tumor weights at the endpoint (* P < 0.05; *** P < 0.001). j Kaplan-Meier survival analysis of patients with different risk scores by CDK7 protein level and tumor-infiltrating lymphocyte (TIL) scores in cohort II. High CDK7 protein level and low TIL scores were defined as two risk factors. Patients were stratified into three risk groups with different survival outcomes as follows: low-risk group without any risk factors = low CDK7 protein level and high TIL score ( n = 64); medium risk group with only one risk factor ( n = 126); high-risk group with two risk factors = high CDK7 protein level and low TIL score ( n = 32) (* P < 0.05). k Kaplan-Meier survival analysis of patients with different risk scores by CDK7 protein level, MYC protein level, and TILs score in cohort II. High CDK7 protein level, high MYC protein level and low TIL score were defined as three risk factors. Patients were stratified into three risk groups with different survival outcomes as follows: low-risk group without any risk factors ( n = 41); medium risk group with only one risk factor ( n = 97); high-risk group with at least two risk factors ( n = 84) (** P < 0.01). l Schematic illustration showing the CDK7-p38α-MYC axis dependent regulation of PD-L1 and the function of this signaling in NSCLC. The solid arrows represent strong processes; the dashed arrows represent very weak processes after CDK7 inhibition

    Article Snippet: CD8 + T cells were depleted in a subset of mice by intraperitoneal injection of CD8a depletion antibody (BioXcell, 2.43 clone), 3 times per week at 150 μg per animal.

    Techniques: Inhibition, Quantitation Assay, Enzyme-linked Immunosorbent Assay

    Specifications of tracers

    Journal: Theranostics

    Article Title: CD4 + and CD8a + PET imaging predicts response to novel PD-1 checkpoint inhibitor: studies of Sym021 in syngeneic mouse cancer models

    doi: 10.7150/thno.37513

    Figure Lengend Snippet: Specifications of tracers

    Article Snippet: Mice bearing subcutaneous CT26 tumors (150-200 mm 3 ) were treated for three consecutive days with intraperitoneal injections of either saline, 300 μg of CD8a + depleting antibody clone 2.43, a full length rat-anti-mouse IgG2b (#BE0061, BioXcell), or 300 μg DFO-CD8a precursor (N=5/group).

    Techniques: Activity Assay, Injection

    Development and optimization of radiolabeled F(ab)'2 fragments for PET imaging . (A) Schematic illustration of preparation of F(ab)'2 fragments from full length antibodies using FaBRICATOR (IdeS protease). (B) SDS-page of non-digested (lane 2+6), crude antibody mixture (lane 3+7) and purified F(ab)'2 (lane 4+8) and Fc fragments (lane 5+9) of anti-mouse CD4 and anti-mouse CD8a. (C) Representative HPLC chromatogram of crude anti-mouse CD8a antibody mixture used for preparative purification. (D) Representative HPLC chromatogram of 89 Zr-DFO-CD8a after PD10 purification at end-of-synthesis. (E) Ex vivo biodistribution 72 hours post-injection of 89 Zr-DFO-CD8a in major organs and (F) CT26 tumors with increasing doses of unlabeled CD8a-F(ab)'2 measured by gamma counting and expressed as %ID/g (N=3/dose). (G) Tumor-to-muscle and (H) tumor-to-blood ratio of uptake quantified by gamma counting 72 hours post-injection of 89 Zr-DFO-CD8a (N=3/dose). (I) Image-derived biodistribution of dose-optimized mean 89 Zr-DFO-CD8a uptake in major organs and tumor based on ROI analysis and expressed as mean %ID/g in CT26 tumor-bearing mice over the imaging time-course (N=3). Data are presented as mean ± SEM. Ab: antibody; ALN: axillary lymph node; CLN: cervical lymph node; ILN: inguinal lymph node; %ID/g: % injected dose per gram tissue.

    Journal: Theranostics

    Article Title: CD4 + and CD8a + PET imaging predicts response to novel PD-1 checkpoint inhibitor: studies of Sym021 in syngeneic mouse cancer models

    doi: 10.7150/thno.37513

    Figure Lengend Snippet: Development and optimization of radiolabeled F(ab)'2 fragments for PET imaging . (A) Schematic illustration of preparation of F(ab)'2 fragments from full length antibodies using FaBRICATOR (IdeS protease). (B) SDS-page of non-digested (lane 2+6), crude antibody mixture (lane 3+7) and purified F(ab)'2 (lane 4+8) and Fc fragments (lane 5+9) of anti-mouse CD4 and anti-mouse CD8a. (C) Representative HPLC chromatogram of crude anti-mouse CD8a antibody mixture used for preparative purification. (D) Representative HPLC chromatogram of 89 Zr-DFO-CD8a after PD10 purification at end-of-synthesis. (E) Ex vivo biodistribution 72 hours post-injection of 89 Zr-DFO-CD8a in major organs and (F) CT26 tumors with increasing doses of unlabeled CD8a-F(ab)'2 measured by gamma counting and expressed as %ID/g (N=3/dose). (G) Tumor-to-muscle and (H) tumor-to-blood ratio of uptake quantified by gamma counting 72 hours post-injection of 89 Zr-DFO-CD8a (N=3/dose). (I) Image-derived biodistribution of dose-optimized mean 89 Zr-DFO-CD8a uptake in major organs and tumor based on ROI analysis and expressed as mean %ID/g in CT26 tumor-bearing mice over the imaging time-course (N=3). Data are presented as mean ± SEM. Ab: antibody; ALN: axillary lymph node; CLN: cervical lymph node; ILN: inguinal lymph node; %ID/g: % injected dose per gram tissue.

    Article Snippet: Mice bearing subcutaneous CT26 tumors (150-200 mm 3 ) were treated for three consecutive days with intraperitoneal injections of either saline, 300 μg of CD8a + depleting antibody clone 2.43, a full length rat-anti-mouse IgG2b (#BE0061, BioXcell), or 300 μg DFO-CD8a precursor (N=5/group).

    Techniques: Imaging, SDS Page, Purification, Ex Vivo, Injection, Derivative Assay

    Specificity of 89 Zr-DFO-CD8a in depleted, antigen-negative and tumor-bearing mice. (A) Representative dot plots of median fluorescent intensity of CD4-FITC (y-axis) and CD8a-BV711 (x-axis) measured by flow cytometric analysis of blood, spleen and tumors of control (N=2), CD8a + depleted (N=2) and DFO-CD8a treated (N=2) CT26 tumor-bearing mice. (B) Depletion with CD8a 2.43 mAb reduced the percentage of CD45 + CD8a + cells in blood, spleen and tumor whereas DFO-CD8a precursor did not change CD45 + CD8a + populations (N=2/group). (C) Representative axial PET/CT images 24 hours post-injection of 89 Zr-DFO-CD8a in control, CD8a + depleted and NMRI nude (antigen-negative) mice. Arrows designate the spleen. (D) Ex vivo biodistribution of 89 Zr-DFO-CD8a and 89 Zr-DFO-IgG2b (isotype control) in lymphoid tissue. Mean 89 Zr-DFO-CD8a uptake was significantly reduced in CD8a + depleted mice (N=3/group). (E) The 89 Zr-DFO-CD8a tumor-to-blood ratio was lowered in CD8a + depleted mice compared to control mice (p=0.011) (N=3/group). The 89 Zr-DFO-IgG2b (isotype control) tumor-to-blood ratio was significantly different than the 89 Zr-DFO-CD8a tumor-to-blood ratio in control (p=0.0001) and CD8a + depleted mice (p=0.0024). (F) Autoradiography of tumors showed increased 89 Zr-DFO-CD8a uptake in tumors subjected to fractionated external radiation therapy (XRT, 3x2Gy). (G) XRT (3x2Gy) increased the mean 89 Zr-DFO-CD8a uptake in tumors (p=0.0006) and spleens (p=0.0022) of CT26 tumor-bearing mice (N=16/group) that was confirmed by (H) flow cytometric analysis of CD45 + CD8a + cells (N=6/group). Data are presented as mean ± SEM and the significance levels are indicated by asterisks (*). *=p<0.05, **=p<0.01, ***=p<0.001, ****=p<0.0001, ns=no significance. ALN: axillary lymph node; CLN: cervical lymph node; ILN: inguinal lymph node; XRT: external radiation therapy; %ID/g: % injected dose per gram tissue.

    Journal: Theranostics

    Article Title: CD4 + and CD8a + PET imaging predicts response to novel PD-1 checkpoint inhibitor: studies of Sym021 in syngeneic mouse cancer models

    doi: 10.7150/thno.37513

    Figure Lengend Snippet: Specificity of 89 Zr-DFO-CD8a in depleted, antigen-negative and tumor-bearing mice. (A) Representative dot plots of median fluorescent intensity of CD4-FITC (y-axis) and CD8a-BV711 (x-axis) measured by flow cytometric analysis of blood, spleen and tumors of control (N=2), CD8a + depleted (N=2) and DFO-CD8a treated (N=2) CT26 tumor-bearing mice. (B) Depletion with CD8a 2.43 mAb reduced the percentage of CD45 + CD8a + cells in blood, spleen and tumor whereas DFO-CD8a precursor did not change CD45 + CD8a + populations (N=2/group). (C) Representative axial PET/CT images 24 hours post-injection of 89 Zr-DFO-CD8a in control, CD8a + depleted and NMRI nude (antigen-negative) mice. Arrows designate the spleen. (D) Ex vivo biodistribution of 89 Zr-DFO-CD8a and 89 Zr-DFO-IgG2b (isotype control) in lymphoid tissue. Mean 89 Zr-DFO-CD8a uptake was significantly reduced in CD8a + depleted mice (N=3/group). (E) The 89 Zr-DFO-CD8a tumor-to-blood ratio was lowered in CD8a + depleted mice compared to control mice (p=0.011) (N=3/group). The 89 Zr-DFO-IgG2b (isotype control) tumor-to-blood ratio was significantly different than the 89 Zr-DFO-CD8a tumor-to-blood ratio in control (p=0.0001) and CD8a + depleted mice (p=0.0024). (F) Autoradiography of tumors showed increased 89 Zr-DFO-CD8a uptake in tumors subjected to fractionated external radiation therapy (XRT, 3x2Gy). (G) XRT (3x2Gy) increased the mean 89 Zr-DFO-CD8a uptake in tumors (p=0.0006) and spleens (p=0.0022) of CT26 tumor-bearing mice (N=16/group) that was confirmed by (H) flow cytometric analysis of CD45 + CD8a + cells (N=6/group). Data are presented as mean ± SEM and the significance levels are indicated by asterisks (*). *=p<0.05, **=p<0.01, ***=p<0.001, ****=p<0.0001, ns=no significance. ALN: axillary lymph node; CLN: cervical lymph node; ILN: inguinal lymph node; XRT: external radiation therapy; %ID/g: % injected dose per gram tissue.

    Article Snippet: Mice bearing subcutaneous CT26 tumors (150-200 mm 3 ) were treated for three consecutive days with intraperitoneal injections of either saline, 300 μg of CD8a + depleting antibody clone 2.43, a full length rat-anti-mouse IgG2b (#BE0061, BioXcell), or 300 μg DFO-CD8a precursor (N=5/group).

    Techniques: Control, Positron Emission Tomography-Computed Tomography, Injection, Ex Vivo, Autoradiography

    Characterization of syngeneic mouse models by CD4 + and CD8a + subsets. (A) Representative sections from immunohistochemical (IHC) staining of CD4 + and CD8a + , and hematoxylin and eosin (HE) staining in a tumor model with low (B16F10) and high (Sa1N) intensity staining (left panel). Representative dot plots of median fluorescent intensity of CD4-FITC (y-axis) and CD8a-PerCP-Cy5.5 (x-axis) of flow cytometric analysis of CD4 + and CD8a + cells in a tumor model with low (B16F10) and high (Sa1N) intensity staining (right panel). (B) Agreement between flow cytometric and IHC analysis of CD4 + and (C) CD8a + subsets revealed clustering of tumor types into hot (red) and cold (blue) areas. N=6/model for flow cytometry, N=3/model for IHC. HE: hematoxylin and eosin; IHC: immunohistochemistry.

    Journal: Theranostics

    Article Title: CD4 + and CD8a + PET imaging predicts response to novel PD-1 checkpoint inhibitor: studies of Sym021 in syngeneic mouse cancer models

    doi: 10.7150/thno.37513

    Figure Lengend Snippet: Characterization of syngeneic mouse models by CD4 + and CD8a + subsets. (A) Representative sections from immunohistochemical (IHC) staining of CD4 + and CD8a + , and hematoxylin and eosin (HE) staining in a tumor model with low (B16F10) and high (Sa1N) intensity staining (left panel). Representative dot plots of median fluorescent intensity of CD4-FITC (y-axis) and CD8a-PerCP-Cy5.5 (x-axis) of flow cytometric analysis of CD4 + and CD8a + cells in a tumor model with low (B16F10) and high (Sa1N) intensity staining (right panel). (B) Agreement between flow cytometric and IHC analysis of CD4 + and (C) CD8a + subsets revealed clustering of tumor types into hot (red) and cold (blue) areas. N=6/model for flow cytometry, N=3/model for IHC. HE: hematoxylin and eosin; IHC: immunohistochemistry.

    Article Snippet: Mice bearing subcutaneous CT26 tumors (150-200 mm 3 ) were treated for three consecutive days with intraperitoneal injections of either saline, 300 μg of CD8a + depleting antibody clone 2.43, a full length rat-anti-mouse IgG2b (#BE0061, BioXcell), or 300 μg DFO-CD8a precursor (N=5/group).

    Techniques: Immunohistochemical staining, Immunohistochemistry, Staining, Flow Cytometry

    Summarized IHC, flow cytometry, PET imaging and efficacy data.

    Journal: Theranostics

    Article Title: CD4 + and CD8a + PET imaging predicts response to novel PD-1 checkpoint inhibitor: studies of Sym021 in syngeneic mouse cancer models

    doi: 10.7150/thno.37513

    Figure Lengend Snippet: Summarized IHC, flow cytometry, PET imaging and efficacy data.

    Article Snippet: Mice bearing subcutaneous CT26 tumors (150-200 mm 3 ) were treated for three consecutive days with intraperitoneal injections of either saline, 300 μg of CD8a + depleting antibody clone 2.43, a full length rat-anti-mouse IgG2b (#BE0061, BioXcell), or 300 μg DFO-CD8a precursor (N=5/group).

    Techniques: Flow Cytometry, Imaging

    Experimental design and PET imaging of 89 Zr-DFO-CD4 and 89 Zr-DFO-CD8a in a panel of syngeneic mouse models. (A) Overview of the timing of tumor inoculation, 89 Zr-DFO-CD4 and 89 Zr-DFO-CD8a injections, PET imaging and therapy dosing. Sym021 (10 mg/kg) was dosed 6 times over two weeks and efficacy monitored until humane endpoints were reached. (B) Maximum 89 Zr-DFO-CD4 and (C) 89 Zr-DFO-CD8a uptake in syngeneic mouse models quantified from PET ROI analysis of tumors and expressed as %ID/g 24 hours post-injection of tracer ranked from low (left) to high (right). Tumor-to-heart ratios of the maximum (D) 89 Zr-DFO-CD4 and (E) 89 Zr-DFO-CD8a uptake quantified from PET ROI analysis and expressed as %ID/g 24 hours post-injection of tracer ranked from low (left) to high (right). (F) Representative coronal maximum intensity projection PET images of 89 Zr-DFO-CD4 (top panel) and 89 Zr-DFO-CD8a (bottom panel) for each model. The PET acquisition time was 300 seconds. White circles designate the tumor. Data are presented as mean ± SEM. %ID/g: % injected dose per gram tissue.

    Journal: Theranostics

    Article Title: CD4 + and CD8a + PET imaging predicts response to novel PD-1 checkpoint inhibitor: studies of Sym021 in syngeneic mouse cancer models

    doi: 10.7150/thno.37513

    Figure Lengend Snippet: Experimental design and PET imaging of 89 Zr-DFO-CD4 and 89 Zr-DFO-CD8a in a panel of syngeneic mouse models. (A) Overview of the timing of tumor inoculation, 89 Zr-DFO-CD4 and 89 Zr-DFO-CD8a injections, PET imaging and therapy dosing. Sym021 (10 mg/kg) was dosed 6 times over two weeks and efficacy monitored until humane endpoints were reached. (B) Maximum 89 Zr-DFO-CD4 and (C) 89 Zr-DFO-CD8a uptake in syngeneic mouse models quantified from PET ROI analysis of tumors and expressed as %ID/g 24 hours post-injection of tracer ranked from low (left) to high (right). Tumor-to-heart ratios of the maximum (D) 89 Zr-DFO-CD4 and (E) 89 Zr-DFO-CD8a uptake quantified from PET ROI analysis and expressed as %ID/g 24 hours post-injection of tracer ranked from low (left) to high (right). (F) Representative coronal maximum intensity projection PET images of 89 Zr-DFO-CD4 (top panel) and 89 Zr-DFO-CD8a (bottom panel) for each model. The PET acquisition time was 300 seconds. White circles designate the tumor. Data are presented as mean ± SEM. %ID/g: % injected dose per gram tissue.

    Article Snippet: Mice bearing subcutaneous CT26 tumors (150-200 mm 3 ) were treated for three consecutive days with intraperitoneal injections of either saline, 300 μg of CD8a + depleting antibody clone 2.43, a full length rat-anti-mouse IgG2b (#BE0061, BioXcell), or 300 μg DFO-CD8a precursor (N=5/group).

    Techniques: Imaging, Injection

    89 Zr-DFO-CD4 PET predicts Sym021 treatment outcome in syngeneic mouse models. (A) The tumor growth inhibition (TGI) from day 0 til day 10 in Sym021 treated mice (10 mg/kg) relative to the growth of the control group expressed as % correlated with the maximum tumor-to-heart ratio of 89 Zr-DFO-CD4 (p=0.0002, top panel) and 89 Zr-DFO-CD8a (p=0.0354, bottom panel) in individual mice across all models (N=35/tracer). (B) Average TGI until day 10 relative to start of therapy with 10 mg/kg Sym021 (day 0) expressed as % for each model plotted against the average maximum tumor-to-heart ratio of 89 Zr-DFO-CD4 (top panel) and 89 Zr-DFO-CD8a (bottom panel) for each model (N=35/tracer, N=5/model). (C) Mean and maximum tumor-to-heart ratio of 89 Zr-DFO-CD4 (top panel) and 89 Zr-DFO-CD8a (bottom panel) in responders (Rs) and non-responders (N-Rs) across all models (N=35/tracer). Responders (N=5) had higher maximum 89 Zr-DFO-CD4 tumor-to-heart ratio than all non-responders (N=30) (p=0.0306, top panel). (D) Mean and maximum tumor-to-heart ratio of 89 Zr-DFO-CD4 (top panel) and 89 Zr-DFO-CD8a (bottom panel) in responders and non-responders of the models where responders were found. Responders (N=5) had higher maximum 89 Zr-DFO-CD4 tumor uptake than the non-responders of the models were responders were found (N=10) (p=0.0097, top panel). (E) Stratification of mice across all models based on their maximum tumor-to-heart ratio of 89 Zr-DFO-CD4 (top panel) and 89 Zr-DFO-CD8a (bottom panel) resulted in improved overall survival of mice with a maximum 89 Zr-DFO-CD4 tumor-to-heart ratio >9 (N=5) compared to the <9 group (N=30) (p=0.0018). All tumor and heart uptake values of 89 Zr-DFO-CD4 and 89 Zr-DFO-CD8a were derived from PET ROI analysis and expressed as %ID/g. Data are presented as mean ± SEM and the significance levels are indicated by asterisks (*). * = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001, ns = no significance. N-Rs: non-responders; ROI: region of interest; Rs: responders; %ID/g: % injected dose per gram tissue.

    Journal: Theranostics

    Article Title: CD4 + and CD8a + PET imaging predicts response to novel PD-1 checkpoint inhibitor: studies of Sym021 in syngeneic mouse cancer models

    doi: 10.7150/thno.37513

    Figure Lengend Snippet: 89 Zr-DFO-CD4 PET predicts Sym021 treatment outcome in syngeneic mouse models. (A) The tumor growth inhibition (TGI) from day 0 til day 10 in Sym021 treated mice (10 mg/kg) relative to the growth of the control group expressed as % correlated with the maximum tumor-to-heart ratio of 89 Zr-DFO-CD4 (p=0.0002, top panel) and 89 Zr-DFO-CD8a (p=0.0354, bottom panel) in individual mice across all models (N=35/tracer). (B) Average TGI until day 10 relative to start of therapy with 10 mg/kg Sym021 (day 0) expressed as % for each model plotted against the average maximum tumor-to-heart ratio of 89 Zr-DFO-CD4 (top panel) and 89 Zr-DFO-CD8a (bottom panel) for each model (N=35/tracer, N=5/model). (C) Mean and maximum tumor-to-heart ratio of 89 Zr-DFO-CD4 (top panel) and 89 Zr-DFO-CD8a (bottom panel) in responders (Rs) and non-responders (N-Rs) across all models (N=35/tracer). Responders (N=5) had higher maximum 89 Zr-DFO-CD4 tumor-to-heart ratio than all non-responders (N=30) (p=0.0306, top panel). (D) Mean and maximum tumor-to-heart ratio of 89 Zr-DFO-CD4 (top panel) and 89 Zr-DFO-CD8a (bottom panel) in responders and non-responders of the models where responders were found. Responders (N=5) had higher maximum 89 Zr-DFO-CD4 tumor uptake than the non-responders of the models were responders were found (N=10) (p=0.0097, top panel). (E) Stratification of mice across all models based on their maximum tumor-to-heart ratio of 89 Zr-DFO-CD4 (top panel) and 89 Zr-DFO-CD8a (bottom panel) resulted in improved overall survival of mice with a maximum 89 Zr-DFO-CD4 tumor-to-heart ratio >9 (N=5) compared to the <9 group (N=30) (p=0.0018). All tumor and heart uptake values of 89 Zr-DFO-CD4 and 89 Zr-DFO-CD8a were derived from PET ROI analysis and expressed as %ID/g. Data are presented as mean ± SEM and the significance levels are indicated by asterisks (*). * = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001, ns = no significance. N-Rs: non-responders; ROI: region of interest; Rs: responders; %ID/g: % injected dose per gram tissue.

    Article Snippet: Mice bearing subcutaneous CT26 tumors (150-200 mm 3 ) were treated for three consecutive days with intraperitoneal injections of either saline, 300 μg of CD8a + depleting antibody clone 2.43, a full length rat-anti-mouse IgG2b (#BE0061, BioXcell), or 300 μg DFO-CD8a precursor (N=5/group).

    Techniques: Inhibition, Control, Derivative Assay, Injection

    (A) Weights of subcutaneous LLC tumors from 1-month mock and Aza + ITF-2357 treated mice in the presence of CD8a-depleting antibody (n = 7 mock and n = 5 treated mice). NS, non-significant p value calculated by two tail t test.

    Journal: Cell

    Article Title: Epigenetic Therapy Ties MYC Depletion to Reversing Immune Evasion and Treating Lung Cancer

    doi: 10.1016/j.cell.2017.10.022

    Figure Lengend Snippet: (A) Weights of subcutaneous LLC tumors from 1-month mock and Aza + ITF-2357 treated mice in the presence of CD8a-depleting antibody (n = 7 mock and n = 5 treated mice). NS, non-significant p value calculated by two tail t test.

    Article Snippet: CD8a depletion antibody (BioXcell, 2.43 clone), 3 times per week at 150ug per animal, see LLC CD8 depletion model for details about the treatment schema.

    Techniques:

    KEY RESOURCES TABLE

    Journal: Cell

    Article Title: Epigenetic Therapy Ties MYC Depletion to Reversing Immune Evasion and Treating Lung Cancer

    doi: 10.1016/j.cell.2017.10.022

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: CD8a depletion antibody (BioXcell, 2.43 clone), 3 times per week at 150ug per animal, see LLC CD8 depletion model for details about the treatment schema.

    Techniques: Western Blot, Plasmid Preparation, Recombinant, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, Amplification, Expressing, Software