Inhibition of T-cell proliferation is the most common approach to assess human myeloid-derived suppressor cell (MDSC) functions. However, diverse methodologies hinder the comparison of results obtained in different laboratories. In this chapter, we present a T-cell proliferation assay procedure based ...
18kAccesses 10Altmetric Metrics Abstract The antitumor efficacy of genetically engineered ‘living drugs’, including chimeric antigen receptor and T-cell receptor T cells, is influenced by their activation, proliferation, inhibition, and exhaustion. A sensitive and reproducible cytotoxicity assay that colle...
Cell proliferation assay Fresh prepared T-cells were washed by PBS twice and stained with 2 uM CellTrace Voilet (Invitrogen) in PBS for 10 min, and incubated in complete medium for another 20 min to quench residual dye. After two wash with complete medium, cells were activated at pHe...
,“non-Treg cells”) isolated from the mPEG-MLR, also exhibited significant, though weaker, inhibition of allogeneic CD8+ T cell proliferation (roughly comparable to CD25+ cells prepared from naive mice). This alloinhibition was likely due to the existence of non-CD25+ regulatory T cell ...
In vitro proliferation assay of murine colon cancer cells For cell proliferation assay, cells were seeded in flat-bottom 96 well plates at a density of 3000 cells/well. Cell proliferating rate was determined by counting the cells with the counting kit-8 (APExBIO) at 0, 24, 48, and 72 h...
The inhibition of protein tyrosine phosphatases 1B (PTP1B) and N2 (PTPN2) has emerged as an exciting approach for bolstering T cell anti-tumor immunity. ABBV-CLS-484 is a PTP1B/PTPN2 inhibitor in clinical trials for solid tumors. Here we have explored th
In two other models of antitumor immunity, it was proposed that Tregs diminished CD8+ T-cell function by consuming IL-2 [42] or by preventing cytotoxic granule release [43]. In a model of autoimmunity, other authors proposed that DCs were central to Treg inhibition in vivo and attenuated ...
Inhibition of Rho-Kinase downregulates Th17 cells and ameliorates hepatic fibrosis by Schisto- soma japonicum infection. Cells. 2019;8:1262. 4. Carson JP, Ramm GA, Robinson MW, McManus DP, Gobert GN. Schisto- some-induced fibrotic disease: the role of hepatic stellate cells. Trends Parasitol...
We hypothesized that GPR171 is an understudied functional receptor for T cells. To test that, we utilized neuropeptide BigLEN, a known natural ligand for GPR17121, to amplify GPR171 signal in a human T-cell proliferation assay. Under the stimulation of a human CD3 mAb (clone OKT3), the pr...
Proliferation was assessed by a CFDA dilution assay. e CD25 and CD69 expression of T cell subpopulations after 24 h of coculture (n = 5 individual T cell donors). CD3+ PBTs were categorized into effector memory (EM, CD45RA−CCR7−), central memory (CM, CD45RA−CCR7+), ...