Visit Sartorius at ECI 2018, booth #39, Amsterdam from 3rd to 5th September to learn the latest on the Intellicyt® iQue Screener PLUS and the IncuCyte® S3 Live-Cell Analysis System.
Our innovative cell analysis systems provide specialized instrumentation, software and reagents to accelerate discovery and development of novel therapeutics and provide new insight into the mechanisms of disease at a speed, depth and scale not achievable with conventional cell analysis techniques.
While at ECI, please visit our poster presentation.
A Rapid, High Throughput Multiplex Assay that Measures T-Cell Activation, Cytokine Secretion, and Identifies T-cell Subsets from Multiple Donors
Presenter: John O’Rourke, Ph.D – MBA, Assay Development Manager Poster Session: P.B4.03 Session Title: T-cell activation and exhaustion – Part 3 Date and Time: Wednesday, 05.09.2018 10:15-11:45 (Guided Poster Walk) Location: Poster Area (Exhibition Hall) Authors: Zhaoping Liu and John O’Rourke
Intellicyt Corporation, Part of the Sartorius Group
The identification of T-cell subsets and assessing their ex vivo activation is a key step in the adoptive cell therapy process. To facilitate this workflow, we developed a high throughput flow cytometry-based, multiplexed assay to measure T cell activation in different T-cell subsets. PBMCs from multiple donors were profiled prior to and after T cell enrichment with CD3/CD28 magnetic beads. Purified T cells were cultured in the presence of the beads and media containing IL-2 for 3 days and cell phenotypes and secreted cytokines were analyzed. For each sample well, 11 cytokines and 13 cellular endpoints were measured each day over the course of a three-day activation protocol including quantifying cells expressing the activation markers CD69, CD25 and HLA-DR. A 96-well sample plate contained T cells from 12 different donors, with each sample analyzed in quadruplicate using two different sample dilutions. The plates were read on the iQue Screener Plus and the high-content data was analyzed using the integrated ForeCyt software. The results indicate a time course dependent, donor to donor variation in T-cell activation. These high throughput, large-scale flow cytometry studies provide extensive T cell activation and subset profiles from multiple donors with assay times of ~15 minutes. The adoption of these assays can provide actionable data to optimize adoptive cell therapy culture conditions and improve manufacturing protocols.
