Journal: Cancers

Article Title: Insulin Resistance in Women Correlates with Chromatin Histone Lysine Acetylation, Inflammatory Signaling, and Accelerated Aging

doi: 10.3390/cancers16152735

Figure Lengend Snippet: Increased senescent T-cells in insulin-resistant ( IR ) women versus metabolically healthy controls ( HC ). Flow cytometry was used to test peripheral white blood cells (WBCs) from 9 insulin-resistant women (HbA1c = 5.7–6.3) vs. 13 metabolically healthy women (HbA1 < 5.7) for beta-galactosidase (SA-beta-gal) activity, a marker of senescence. There was a significant increase in the percentage of senescent CD4+- and CD8+-positive T-cells in insulin-resistant women vs. healthy women; ** p = 0.0046 and * p = 0.0245, respectively.

Article Snippet: PBMCs (1.0 × 10 6 ) were prepared for flow cytometry by staining for the immune cell subsets, CD45+CD3+CD4+ T-cells, and CD45+CD3+CD8+ T-cells using anti-human CD45 V500 (BD Biosciences, Frankliin Lakes, NJ, USA, Catalog No. 560777), anti-human CD3 Super Bright 600 (ThermoFisher Scientific, Waltham, MA, USA, Catalog No. 63-0037-42), anti-human CD4 PE-Cy7 (BD Biosciences, Catalog No. 348789), and anti-human CD8 cFlourYG610 (Cytek Biosciences, Fremont, CA, USA, Catalog No. R7-20245).

Techniques: Metabolic Labelling, Flow Cytometry, Activity Assay, Marker

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: Targeted mutation of the Hif1a gene increased PDL1 levels in donor T cells and non-tumor host tissues via IFNγ-dependent pathway Flow cytometry analysis of spleen samples from WT or Hif1a −/− T cell recipients 14 days after transplantation. (A) The percentage of WT and Hif1a −/− donor T cell in splenocytes of 14 days post-transplantation mice, shown as mean ± SEM. Data are representative of 3 independent experiments. (B) Frequency of IFNγ + T cells in spleen. Splenocytes from recipients reconstituted with WT or Hif1a −/− T cells were stimulated with PMA + ionomycin for 4 h, and IFNγ expression was determined by intracellular staining. The summarized data are shown as mean ± SEM for one experiment and are representative of 3 independent experiments. (C) The mean fluorescence intensity (MFI) of PDL1 staining is plotted for CD4 (left) and CD8 (right) donor T cells from spleens of recipient mice on day 14. (D) The CD4/CD8 ratio of donor WT and Hif1a −/− T cells in splenocytes of 14 days post-transplantation mice was measured and summarized, shown as mean ± SEM. Data are shown for one experiment and are representative of at least 3 independent experiments. (E) BALB/c mice received 5 × 10 6 CD45.1 T cell-depleted BM cells + 3 × 10 5 WT or Hif1a −/− T cells and were subsequently treated with anti-mouse IFNγ (XMG1.2) or isotype control IgG antibodies (vehicle). Representative immunofluorescence staining for CD3 and PDL1 in the liver and S.G. tissues from mice receiving different treatments. (F) Kaplan-Meier survival curves are shown for the four groups of mice. The data are representative of three experiments. (G) H&E-stained tissues are shown in representative images for each of four groups of mice, highlighting major pathological findings. (H) Summary of histological scoring for the organs in each group depicted in (H). Scoring criteria are described in the .

Article Snippet: PE-Cy7 conjugated anti–human CD4 , eBioscience , 25-0047-42; 25-0047-42.

Techniques: Mutagenesis, Flow Cytometry, Transplantation Assay, Expressing, Staining, Fluorescence, Control, 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: PE-Cy7 conjugated anti–human CD4 , eBioscience , 25-0047-42; 25-0047-42.

Techniques: Recombinant, Software