Additionally, while the data that exist to train these models comes through high-throughput methods such as tetramer sorting21,22 or T cell stimulation assays31, these methods often introduce some level of noise due to non-specific binding or stimulation. Unfortunately, gold standard methods that ...
T cell stimulation assay Cells were digested from the limbs of arthritic mice harvested at ZT6 or ZT18. Cells were counted (BioRad TC20 automated cell counter) in the presence of trypan blue. Samples were washed and re-suspended in stimulation cocktail (containing 0.1 μg/mL ionomycin, ...
Cell purity was generally 92-97%, as determined by flow cytometry. Intracellular staining for FoxP3 was performed using the anti-mouse/rat FoxP3 staining kit from eBioscience (San Diego, CA). In vitro stimulation assays Spleen cells (2 × 105/well) were incubated for 72 hours in flat ...
This chapter provides an overview of cytometric cytokine secretion assay used for the detection and isolation of cytokine-secreting T cells. The cytokine secretion assay is useful for the detection and isolation of viable antigen-specific T cells after a short restimulation with specific antigen in ...
3.5.2T cell proliferation T cellproliferation in response to stimulation by a peptide–MHC complex can be measured by (1) the incorporation of the radioactive nucleotide tritiatedthymidineinto the DNA of dividing but not resting cells or (2) thedilutionof afluorescent dye,carboxyfluoresceinsuccinimidy...
Besides, S100A9+CD14+ monocytes exhibit high expression of PD-L1, enabling its interaction with PD-1 on T cells and the subsequent inhibition of T cell proliferation and T-cell cytotoxicity. This study provides novel insights into the existence of a previously unrecognized circulating monocyte ...
Non-coding disease-associated variants are enriched within cell type-specific open chromatin regions, especially regulatory elements such as promoters and enhancers [7, 11, 15,16,17,18,19,20,21], and they often impact gene expression in a cell type-specific manner [22,23,24,25,26,27,28,...
First, we conducted the OT-I transgenic mouse model and the CAR-T cell-mediated cytotoxicity assay to investigate the influence of IFI35 on antitumor immunity. CD8+ T cells were isolated from OT-I mice and activated with anti-CD3 (1 µg/mL), anti-CD28 (1 µg/mL), and IL-2 (...
Immune cells must adapt to different environments during the course of an immune response. Here we study the adaptation of CD8+ T cells to the intestinal microenvironment and how this process shapes the establishment of the CD8+ T cell pool. CD8+ T cells
Atherosclerosis is accompanied by a CD4 T cell response to apolipoprotein B (APOB). Major histocompatibility complex class II (MHC-II) tetramers can be used to isolate antigen-specific CD4 T cells by flow sorting. Here, we produce, validate and use an MHC-II tetramer, DRB1*07:01 AP...