What Is Immune Cell Clustering and Proliferation?
During an immune response, activated cells of the immune system, such as T lymphocytes, undergo rapid expansion in order to fight infection or disease. Many interactions also occur between activated immune cells (e.g., T cell interactions with antigen-presenting cells and interactions between T cells themselves). These dynamic changes in cell number and cell-cell interactions are fundamental to regulating the effect and extent of the immune response. Understanding the mechanisms of immune cell activation, proliferation and regulation is key to the development of novel and effective anti-tumor immunotherapeutics.
About The Assays
Introducing the Incucyte® Immune Cell Clustering & Proliferation Assays
Visualize and quantify immune cell interactions and proliferation in real time. Investigate the mechanisms of immune cell activation (proliferation), regulation and differentiation (clustering) inside your incubator and without the need to lift or label your cells.
Real-Time Visualization and Automated Analysis
- Monitor proliferation and cell-cell clustering interactions in real time, without removing your cultures from the incubator
- Automated imaging and analysis – count label-free clusters using intuitive IncuCyte® image analysis software
- Gain phenotypic insight from images and movies
Figure 1. Label-free detection of T cell aggregation. Activation of PBMCs with anti-CD3 antibody and IL-2 induces T cell aggregation or "clustering". User-friendly IncuCyte® software enables accurate quantification of cell clustering over time (blue mask).
Monitor the Full Time Course Inside Your Cell Incubator
- Reveal how treatments affect the kinetics of immune cell clustering and proliferation
- Cultures remain undisturbed and in your incubator throughout the experiment
Figure 2. Measure clustering in real time. Automated time-course analysis reveals treatment effects. Increasing the concentration of anti-CD3 antibody from 0.01 to 10 µg/ml in the presence of IL-2 (10 ng/ml) promotes T cell clustering
No Washing, No Cell Lifting, No Antibody Labeling
- Measure proliferation and clustering without the need for labels
- Set up and walk away – fully automated image capture and analysis
- Compatible with complementary flow cytometry and end-point biochemical measurements – simply harvest cells/supernatants when the assay is complete
Real-time, label-free monitoring of PBMC proliferation.
The Incucyte® Live-Cell Analysis System automatically quantifies proliferation of PBMCs (yellow mask) activated with anti-CD3 antibody and IL-2 over a period of 144 hours. The full time course of PBMC proliferation is shown in the presence of anti-CD3 antibody and IL-2, anti-CD3 antibody alone, and in the absence of activators.
Zumwalde NA, Domae E, Mescher MF, Shimizu Y. ICAM-1-dependent homotypic aggregates regulate CD8 T cell effector function and differentiation during T cell activation. (INSERT LINK) J Immunol, 191(7):3681–93, 2013
Revealing the mechanisms of activated T cell aggregation using IncuCyte ZOOM® real time imaging and cell cluster analysis.
"T cell clusters regulate the tuning of CD8+ T cell function and terminal differentiation. These studies contribute to our knowledge of the necessity of T cell interactions and crosstalk during priming and potentially, via cluster manipulation, we hope to augment vaccine efficacy and anti-tumor immunotherapeutics".
Zumwalde, N. A. The Role of ICAM-1 Mediated T cell:T cell Interactions on CD8+ T cell Effector Function and Differentiation. PhD Dissertation, University of Minnesota (2013)