Journal: Cell Reports Medicine

Article Title: Genetic and pharmaceutical targeting of HIF1α allows combo-immunotherapy to boost graft vs. leukemia without exacerbation graft vs. host disease

doi: 10.1016/j.xcrm.2023.101236

Figure Lengend Snippet: Anti-PD1 and anti-PDL1 antibodies abrogate the anti-GVHD effect of targeted mutation of Hif1a in donor T cells (A) Diagram of the experimental scheme. BALB/c mice received 5 × 10 6 CD45.1 T cell-depleted BM cells + 5 × 10 5 WT or Hif1a −/− T cells. WT and KO T cell recipients were divided to receive treatment with anti-mouse PDL1 (10F.9G2), anti-mouse PD1 (RMP1-14), or isotype control immunoglobulin G (IgG) antibodies (vehicle), as detailed in the . (B) Kaplan-Meier survival curves are shown for the six groups of mice (n = 10–13 mice per group). The data are representative of three experiments. (C) H&E-stained tissues are shown in representative images for each of six groups of mice, highlighting major pathological findings. (D) Summary of histological scoring for the organs in each group depicted in (C). Scoring criteria are described in the . (E) Representative immunofluorescence staining for CD3 and PDL1 in the liver and salivary gland (S.G.) tissues from mice receiving different treatments. (F and G) Representative immunofluorescence images are shown for liver, lung, intestine, skin, and tongue sections from WT or Hif1a −/− T cell recipients. Co-staining of CD3 and PDL1 is shown in (F), and co-staining of CD3 and cleaved caspase-3 (cCasp3) is shown in (G).

Article Snippet: Antibodies for hCD3 (NBP1, Novus), mCD3 (SP7, abcam), cleaved caspase-3 (Asp175, 5A1E), and mPD-L1 (10F.9G2, BioxCell, West Lebanon, NH) were used for immunofluorescence.

Techniques: Mutagenesis, Staining, Immunofluorescence

Journal: Cell Reports Medicine

Article Title: Genetic and pharmaceutical targeting of HIF1α allows combo-immunotherapy to boost graft vs. leukemia without exacerbation graft vs. host disease

doi: 10.1016/j.xcrm.2023.101236

Figure Lengend Snippet:

Article Snippet: Antibodies for hCD3 (NBP1, Novus), mCD3 (SP7, abcam), cleaved caspase-3 (Asp175, 5A1E), and mPD-L1 (10F.9G2, BioxCell, West Lebanon, NH) were used for immunofluorescence.

Techniques: Recombinant, Software

Journal: Oncoimmunology

Article Title: Small-molecule PD-L1 inhibitor BMS1166 abrogates the function of PD-L1 by blocking its ER export

doi: 10.1080/2162402X.2020.1831153

Figure Lengend Snippet: BMS1166 partially and specifically inhibited PD-L1 glycosylation. (a) PC9/PD-L1 cells were treated with DMSO, 10 μM BMS1166 or 1 μg/ml tunicamycin (TM) for 17 hr, and the whole cell lysates were collected and incubated with PNGase for 1 hr at 37°C, and then processed to western blot analysis using antibodies as indicated. The anti-GAPDH antibody was used as a control for equal protein loading. (b) PC9/PD-L1 or Jurkat/PD-1 cells were treated with DMSO, 10 μM BMS1166 or indicated concentration of TM for 17 hr. Total cell lysates were collected and processed for western blot using antibodies as indicated. The anti-GAPDH antibody was used as a control for equal protein loading. (c) 4T1 cells were transfected with mPD-L1 plasmids for 24 hr and then incubated with DMSO (NC), 10 μM BMS1166 or 1 μg/ml TM overnight. Total cell lysates were collected and processed for western blot using antibodies as indicated. Anti-α-Tubulin antibody was used as a control for equal protein loading.

Article Snippet: Plasmid: pCMV-hPD-L1, pCMV-hPD-1 and pCMV-mPD-L1-HA were purchased from Sino Biological Inc. pGL4.30-luc-NFAT (# E848A) was purchased from Promega. pCDH-PD-L1-WT, N200Q, N219Q, N192/219Q, N200/219Q or 3NQ-Flag were gifts from Prof. Mien-Chie Hung (The University of Texas, Houston, USA).

Techniques: Incubation, Western Blot, Concentration Assay, Transfection