anti mouse cd8 α ab  (Bio X Cell)

Journal: The Journal of Clinical Investigation

Article Title: PPP2R2A insufficiency enhances PD-L1 immune checkpoint blockade efficacy in lung cancer through cGAS-STING activation

doi: 10.1172/JCI193354

Figure Lengend Snippet: ( A–D ) Representative Western blot analysis showing increased PD-L1 protein levels in mouse (CMT167, LLC) and human (A549, H1299) cells after PPP2R2A KO, with quantification from biological replicates. ( E–G ) PD-L1 induction by PPP2R2A KD is dependent on GSK-3β phosphorylation. ( H–I ) qPCR of PD-L1 mRNA in human and mouse cells. ( J–M ) PD-L1 upregulation by PPP2R2A KD requires STING in CMT167 and A549 cells. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Data are reported as mean ± SD ( n = 3). ( B and H ) A Student t test was used for the data analysis. ( D , I , K and M ) Statistical analysis was conducted using 1-way ANOVA followed by Bonferroni post hoc test for multiple comparisons. shcon, short hairpin RNA control.

Article Snippet: After 9 days, mice were randomized to 1 of the following treatment groups: control (rat IgG2b in vivo isotype control, 100 μg/dose; ICH2243, clone 1-2, Ichorbio); α-PD-L1 alone (anti-mouse PD-L1 in vivo antibody, 100 μg/dose; ICH1086, clone 10F.9G2, Ichorbio), α-PD-L1 + α-CD4 (anti mouse CD4, 200 μg/dose, BP0003, Clone:GK1.5, BioXcell); α-PD-L1 plus α-CD8 (anti-mouse CD8, 200 μg/dose; BP0004, clone 53-6.7, BioXcell); α-PD-L1 plus α-CD4/8 (anti-CD4 plus anti-CD8 antibodies, 200 μg/dose for each antibody); and α-PD-L1 plus α-NK (anti-mouse NK1.1, 200 μg/dose; BP0036, clone PK136, BioXcell).

Techniques: Western Blot, Phospho-proteomics, shRNA, Control

Journal: The Journal of Clinical Investigation

Article Title: PPP2R2A insufficiency enhances PD-L1 immune checkpoint blockade efficacy in lung cancer through cGAS-STING activation

doi: 10.1172/JCI193354

Figure Lengend Snippet: ( A ) Treatment schedule in mice bearing Ppp2r2a + / + or Ppp2r2a +/– tumors. ( B–G ) Tumor growth curves and individual tumor volumes with control or anti–PD-L1 antibody. ( H ) Kaplan–Meier survival analysis. ****, P < 0.0001, Kaplan-Meier analysis was used for overall survival. ( I–J ) Mouse body weights on day 0 and day 25. Data are presented as mean ± SEM. *** P < 0.001, **** P < 0.0001 by 1- or 2-way ANOVA with Bonferroni post hoc test.

Article Snippet: After 9 days, mice were randomized to 1 of the following treatment groups: control (rat IgG2b in vivo isotype control, 100 μg/dose; ICH2243, clone 1-2, Ichorbio); α-PD-L1 alone (anti-mouse PD-L1 in vivo antibody, 100 μg/dose; ICH1086, clone 10F.9G2, Ichorbio), α-PD-L1 + α-CD4 (anti mouse CD4, 200 μg/dose, BP0003, Clone:GK1.5, BioXcell); α-PD-L1 plus α-CD8 (anti-mouse CD8, 200 μg/dose; BP0004, clone 53-6.7, BioXcell); α-PD-L1 plus α-CD4/8 (anti-CD4 plus anti-CD8 antibodies, 200 μg/dose for each antibody); and α-PD-L1 plus α-NK (anti-mouse NK1.1, 200 μg/dose; BP0036, clone PK136, BioXcell).

Techniques: Control

Journal: The Journal of Clinical Investigation

Article Title: PPP2R2A insufficiency enhances PD-L1 immune checkpoint blockade efficacy in lung cancer through cGAS-STING activation

doi: 10.1172/JCI193354

Figure Lengend Snippet: ( A ) Treatment schedule for control antibody or anti–PD-L1 therapy followed by immune profiling. ( B–C ) Flow cytometry of CD45 + tumor-infiltrating immune cells with opt-SNE and FlowSOM clustering. ( D–F ) Representative plots and quantification of NK, Treg, CD4 + , and CD8 + T cell populations. Data are presented as mean ± SEM ( n = 10). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by 1-way ANOVA with Bonferroni post hoc test. ( G ) Schematic summary of TME reprogramming in PPP2R2A-deficient NSCLC under PD-L1 blockade.

Article Snippet: After 9 days, mice were randomized to 1 of the following treatment groups: control (rat IgG2b in vivo isotype control, 100 μg/dose; ICH2243, clone 1-2, Ichorbio); α-PD-L1 alone (anti-mouse PD-L1 in vivo antibody, 100 μg/dose; ICH1086, clone 10F.9G2, Ichorbio), α-PD-L1 + α-CD4 (anti mouse CD4, 200 μg/dose, BP0003, Clone:GK1.5, BioXcell); α-PD-L1 plus α-CD8 (anti-mouse CD8, 200 μg/dose; BP0004, clone 53-6.7, BioXcell); α-PD-L1 plus α-CD4/8 (anti-CD4 plus anti-CD8 antibodies, 200 μg/dose for each antibody); and α-PD-L1 plus α-NK (anti-mouse NK1.1, 200 μg/dose; BP0036, clone PK136, BioXcell).

Techniques: Control, Flow Cytometry

Journal: The Journal of Clinical Investigation

Article Title: PPP2R2A insufficiency enhances PD-L1 immune checkpoint blockade efficacy in lung cancer through cGAS-STING activation

doi: 10.1172/JCI193354

Figure Lengend Snippet: ( A ) Experimental design for immune cell depletion in CMT167 Ppp2r2a +/– tumors treated with control or PD-L1 antibodies. ( B–H ) Tumor growth curves showing loss of therapeutic efficacy upon depletion of CD4 + , CD8 + , or NK cells. ( I–J ) Quantification of tumor volumes and weights on day 16. Data are presented as mean ± SEM ( n = 10). * P < 0.05, ** P < 0.01, **** P < 0.0001 by 1-way or 2-way ANOVA with Bonferroni post hoc test.

Article Snippet: After 9 days, mice were randomized to 1 of the following treatment groups: control (rat IgG2b in vivo isotype control, 100 μg/dose; ICH2243, clone 1-2, Ichorbio); α-PD-L1 alone (anti-mouse PD-L1 in vivo antibody, 100 μg/dose; ICH1086, clone 10F.9G2, Ichorbio), α-PD-L1 + α-CD4 (anti mouse CD4, 200 μg/dose, BP0003, Clone:GK1.5, BioXcell); α-PD-L1 plus α-CD8 (anti-mouse CD8, 200 μg/dose; BP0004, clone 53-6.7, BioXcell); α-PD-L1 plus α-CD4/8 (anti-CD4 plus anti-CD8 antibodies, 200 μg/dose for each antibody); and α-PD-L1 plus α-NK (anti-mouse NK1.1, 200 μg/dose; BP0036, clone PK136, BioXcell).

Techniques: Control, Drug discovery

Journal: The Journal of Clinical Investigation

Article Title: PPP2R2A insufficiency enhances PD-L1 immune checkpoint blockade efficacy in lung cancer through cGAS-STING activation

doi: 10.1172/JCI193354

Figure Lengend Snippet: ( A ) Treatment schedule in mice bearing Ppp2r2a + / + or Ppp2r2a +/– CMT167 tumors. ( B–J ) Tumor growth curves with control or anti–PD-L1 antibody ± IFNAR1 neutralization. ( K–L ) Quantification of tumor volumes and weights at endpoint. Data are presented as mean ± SEM. ( n = 10). ** P < 0.01, **** P < 0.0001 by 1-way or 2-way ANOVA with Bonferroni post hoc test.

Article Snippet: After 9 days, mice were randomized to 1 of the following treatment groups: control (rat IgG2b in vivo isotype control, 100 μg/dose; ICH2243, clone 1-2, Ichorbio); α-PD-L1 alone (anti-mouse PD-L1 in vivo antibody, 100 μg/dose; ICH1086, clone 10F.9G2, Ichorbio), α-PD-L1 + α-CD4 (anti mouse CD4, 200 μg/dose, BP0003, Clone:GK1.5, BioXcell); α-PD-L1 plus α-CD8 (anti-mouse CD8, 200 μg/dose; BP0004, clone 53-6.7, BioXcell); α-PD-L1 plus α-CD4/8 (anti-CD4 plus anti-CD8 antibodies, 200 μg/dose for each antibody); and α-PD-L1 plus α-NK (anti-mouse NK1.1, 200 μg/dose; BP0036, clone PK136, BioXcell).

Techniques: Control, Neutralization

Journal: The Journal of Clinical Investigation

Article Title: PPP2R2A insufficiency enhances PD-L1 immune checkpoint blockade efficacy in lung cancer through cGAS-STING activation

doi: 10.1172/JCI193354

Figure Lengend Snippet: ( A ) Treatment schedule. ( B ) Visualization of CD8 + T cell flow cytometry data obtained from day 7 tumor samples. Tumor samples were collected from indicated groups, and UMAP dimension reduction and FlowSOM clustering were applied to identify clusters with distinctive marker expression patterns. ( C ) Key marker expressions were overlaid onto UMAP space. ( D ) Contour plots were generated to display population dynamics across the groups. ( E ) Representative flow cytometry plot showing CX3CR1 and CD44 expression levels in CD8 + T cells. ( F ) Representative quantification of CD44 + CX3CR1 + CD8 + T cells. ( G ) Working model of PPP2R2A deficiency in ICB response. Data are presented as mean ± SEM. ** P < 0.01, **** P < 0.0001 by Tukey’s multiple comparison test.

Article Snippet: After 9 days, mice were randomized to 1 of the following treatment groups: control (rat IgG2b in vivo isotype control, 100 μg/dose; ICH2243, clone 1-2, Ichorbio); α-PD-L1 alone (anti-mouse PD-L1 in vivo antibody, 100 μg/dose; ICH1086, clone 10F.9G2, Ichorbio), α-PD-L1 + α-CD4 (anti mouse CD4, 200 μg/dose, BP0003, Clone:GK1.5, BioXcell); α-PD-L1 plus α-CD8 (anti-mouse CD8, 200 μg/dose; BP0004, clone 53-6.7, BioXcell); α-PD-L1 plus α-CD4/8 (anti-CD4 plus anti-CD8 antibodies, 200 μg/dose for each antibody); and α-PD-L1 plus α-NK (anti-mouse NK1.1, 200 μg/dose; BP0036, clone PK136, BioXcell).

Techniques: Flow Cytometry, Marker, Expressing, Generated, Comparison