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Immunology 2019
May 9 – 13, 2019
San Diego Convention Center
San Diego, CA 

Booth #1536

Visit our booth at Immunology 2019 to learn about our innovative platforms and products. 

  • IncuCyte® S3 Live-Cell Analysis System and Intellicyt iQue® Screener PLUS are propelling innovation in immunology research. These groundbreaking systems provide “information-rich solutions” that deliver new insights into biological mechanisms at unprecedented speed, depth and scale to accelerate immunologic discovery and reveal the dynamic and complex nature of immune cell biology. 
  • arium® laboratory-grade water purification systems – our laboratory-grade water purification systems feature an inspiring, application-oriented design. They enable you to perform your workflows faster and more reliably, simplifying your daily lab work while ensuring cost-efficient operation over the long term.

At the show, please visit our poster presentations and our block symposium.

Learn more about Immunology 2019

Posters

Title: Development of a high throughput, bead-based flow cytometry panel for T-helper cell cytokine quantitation 

Presentation Type: Poster Session
Session Title: Cytokines Regulating Inflammation
Date: Friday, May 10, 2019
Time: 2:30 PM - 3:45 PM
Room: Exhibit/Poster Hall
Authors: John O’Rourke, Caroline Weldon, Lisa Keenan, and Mark B. Carter, Essen BioScience, Albuquerque, New Mexico, USA.

Upon T-cell activation, naive CD4+ cells differentiate into various types of T-helper (Th) cells, which is influenced by the cytokine milieu present during antigen engagement and is further driven by autocrine mechanisms. Th subsets orchestrate adaptive immune responses to specific pathogens and dysregulation of Th cells are major contributors to autoimmune and allergic diseases. Th subsets can be identified, in part, by their unique cytokine secretion profile. 

To measure cytokine secretion from Th cells, Intellicyt has developed multiplex bead-based panels (QPanels) to quantitate Th1 and Th2 (4-, 6- or 9-plex) and Th1, Th2, & Th17 (7-plex) associated cytokines in cell supernatants.  These miniaturized assays require sample volumes of only 10 µl. Data acquisition is performed using 96 or 384 well plates on the iQue Screener platforms.  Pre-mixed reagents and cytokine standards, and templated data analysis, including event gates, gating strategy, 4 parameter standard curves and sample quantitation are auto-generated to allow for ease of use.  

For proof of concept studies, peripheral blood mononuclear cells (PBMCs) from 3 donors were cultured in media containing human sera and IL-2. Cells were stimulated with anti- CD3/CD28 beads and cytokine secretion was measured 1, 2, and 3 days post-stimulation.  Donor to donor variation and temporal differences in secreted cytokine levels were observed. All donor PBMCs secreted IFNγ, GM-CSF and TNFα by 3 days post-stimulation. Notably, one donor showed a more robust Th2 (IL-4, IL-6, and IL-10) and Th17 (IL-17A) response compared to other donors.  These data highlight the use of QPanels in T cell screening, which allows easy assessment of effector function and the identification of Th subsets.


Title: Real-time visualization and quantification of neutrophil activation and function using live-cell analysis

Presentation Type: Poster Session
Session Title: Technological Innovations I
Date: Saturday, May 11, 2019
Time: 2:30 PM - 3:45 PM
Room: Exhibit/Poster Hall
Authors: N. Bevan, G. Lovell, B. O’Clair, L. Kelsey, C. Szybut, H. Campwala, T. Jackson, N. Holtz, E. Endsley, T. Dale, D. Trezise, Essen BioScience, Welwyn Garden City, Hertfordshire & Ann Arbor, Michigan, USA.

Polymorphonuclear neutrophils (PMNs) have multiple functions during the resolution of inflammation. At the inflammation site, increases in chemokines induce infiltration of PMNs via CXCR receptor activation, resulting in cell clearance via phagocytosis and NETosis. Here we describe characterization of the activation and function of PMNs using IncuCyte® live-cell analysis. 

Changes in cell shape and CD marker expression are known indicators of PMN activation.  Freshly isolated PMNs were seeded in 96-well plates, in the presence of FabFlour-488 labeled CD11b antibody for live-cell immunocytochemistry.  Phase and fluorescence images were captured every 30 min for 6 h with IncuCyte S3.  Cell-by-cell analysis enables individual cell segmentation and yields area, shape (eccentricity) and fluorescent intensity metrics. Subsets of PMNs could be analyzed by classifying on each of these parameters. CXCL8 induced a time- and concentration-dependent increase in eccentricity (EC50 0.63 nM) and concomitant increase in CD11b expression (EC50 0.22 nM).  CXCL8-induced activation was attenuated by an anti-CXCL8 antibody. CXCL1 or CCL2 yielded little or no change in either parameter.

As a follow up, we assessed CXCL8-dependent chemotaxis using IncuCyte ClearView Chemotaxis plates, phagocytotic activity with IncuCyte pHrodo® E. coli Bioparticles® and PMA-induced NETosis (IncuCyte Cytotox Green). In all three cases robust, time-dependent signal changes were observed, consistent with known PMN function.  We conclude that live-cell analysis is a flexible and powerful method for analyzing neutrophil activity, where morphological, protein and functional parameters can be readily quantified and integrated over time.  

Other example data sets for subset analysis of proliferation, cell death and cell cycle measurements as well as immuno-phenotyping will be shared to illustrate the value of this approach. 

Title: Developing a novel multiplexed high throughput flow cytometry based immune assay to screen kinase modulators of primary T cell activation 

Presentation Type: Block Symposium 
Session Title: Technological Innovations II 
Date: Saturday, May 11, 2019 
Time: 9:45 AM - 10:00 AM 
Room: Room 30AB 

Presentation Type: Poster Session 
Session Title: Technological Innovations II 
Date: Saturday, May 11, 2019 
Time: 2:30 PM - 3:45 PM 
Room: Exhibit/Poster Hall 
Authors:  John O’Rourke, Andrea Gomez-Donart, Caroline Weldon, & Zhaoping Liu, Essen BioScience, Albuquerque, NM 87113.

Modulating TCR engagement and signaling using biologics, small molecules or genetic engineering is highly relevant to many therapeutic areas. TCR signal initiation is mediated by cytosolic tyrosine kinases, leading to signal amplification through a network of serine-threonine kinases.  Genetic defects, mutations and other mechanisms leading to increased kinase activity and enhanced T cell activation (TCA) is involved in many autoimmune pathologies, making kinases attractive targets for the direct inhibition of TCA and treatment of autoimmune disease.  

The development of drugs and therapies regulating TCR activity require assays to profile T cell function and cell health.  To address this need, we developed a novel multiplexed TCA assay based on high throughput flow cytometry technology with advanced computational algorithms for data analysis. The assay simultaneously measures T cell phenotype, time-dependent expression of T cell activation markers, cell proliferation and viability and quantitates secreted cytokines from a single 10 l sample using a 96 or 384-well plate format.
 
To illustrate the value of the TCA assay in drug discovery, we performed a phenotypic screen using a 152 kinase inhibitor (KI) small molecule library for their ability to inhibit human primary TCA.  Using the Profile Mapping algorithm in our ForeCyt software, we integrated 11 TCA metrics to identify multiple inhibition pathways.  The Profile Map tool was used to group KI based on their inhibition phenotypes, irrespective of their known targets.  These results illustrate the use of Intellicyt’s technology to deliver solutions for drug and biologics discovery.