Journal: Molecular Therapy. Nucleic Acids

Article Title: Preventing ATP Degradation by ASO-Mediated Knockdown of CD39 and CD73 Results in A2aR-Independent Rescue of T Cell Proliferation

doi: 10.1016/j.omtn.2020.06.020

Figure Lengend Snippet: Impact of A2aR Inhibition on T Cell Proliferation in the Presence or Absence of Extracellular ATP Human T cells were labeled with a proliferation dye, activated with anti-CD2/CD3/CD28 tetrameric antibody complexes, and treated with 5 μM of respective ASOs (or 2.5 μM in the case of the CD39 ASO + CD73 ASO condition) without using a transfection reagent. As mock control, cells were not treated with an ASO. On days 3 and 4, 10 μM of A2a receptor inhibitors (A2aRIs) AZD-4635 or CPI-444, dissolved in DMSO, or DMSO and different doses of ATP, was added to the cells. Proliferation of T cells was analyzed by flow cytometry on day 5 (A), and proliferation indices of CD8 + and CD4 + T cells were calculated (B). (A) Representative results from three technical replicates and two independent experiments are shown. (B) The mean + SD from three technical replicates is depicted. Representative results from two independent experiments are shown.

Article Snippet: A2aRIs AZD-4635 and CPI-444 (MedKoo Biosciences, Morrisville, NC) dissolved in DMSO were added on days 3 and 4 (10 μM).

Techniques: Inhibition, Labeling, Transfection, Control, Flow Cytometry