Application

T Cell Activation

Phenotype and Function Kit

T Cell Activation

The activation of naive T cells by an antigen and costimulatory signals initiates clonal expansion of both CD4+ helper and CD8+ cytotoxic T cells. In addition to T cell proliferation, a variety of signalling pathways are activated, leading to the expression of functional cell surface markers and the release of cytokines.

There is a growing market of therapies (bispecific, checkpoint inhibitor antibodies & CAR-T cells) that focus on using the immune system to target cancer. The ability to monitor and quantify activation profiles allows for a greater understanding of how T cells react to certain stimuli which in turn can provide insight into how well drugs will be able to harness the power of the immune system.

Request a Quote

Assay Concept

Multiplexed approach to measure T cell activation, proliferation and cytokine release in a single well.

Figure 1. Illustration of Human T Cell Activation Cell and Cytokine Profiling Kit assay principles. Different T cell phenotypes are profiled for the expression of 3 activation markers: CD69 (early), CD25 (late), and HLA-DR (even later). The 2 effector cytokines (IFNγ and TNFα) are also quantified using 2-plex QBeads in a sandwich immunoassay format in the same well. Simultaneous measurement of T cell proliferation or encoded target cells is possible, but is not included in this illustration.

Key Advantages

icon check

Gain additional biological insight into T cell activation status in physiologically relevant models

Read more below

icon check

Simultaneously quantify surface protein & cytokine expression

Read more below

icon check

Easy to use and interpret; pre-defined gating strategy

Read more below

icon check

Minimal sample manipulation: low volume, one wash, no dilution, mix and read

Read more below

Gain additional biological insight into T cell activation status in physiologically relevant models

Figure 2. Kinase inhibitor screen highlights variation in human T cell activation profiles.

PBMCs were treated with a panel of Kinase inhibitors (20 µM) for one hour. CD3/CD28 Dynabeads were then added to stimulate activation for 24 hours. 10 µL samples were analysed using the T Cell Activation Cell and Cytokine Profiling Kit and the iQue system.

(A) Plate view of CD4+CD69+ gating strategy. Green box identifies media only control. Red box identifies cyclosporine control. Yellow boxes identify the compounds shown on graphs B-D (1: Ruxolitinib; 2: Bisindolylmaleimide I; 3: AZD 7762).

(B, C) Expression of CD69, CD25 and HLA-DR on CD4+ and CD8+ cells.

(D) Release of IFNγ and TNFα cytokines from PBMCs.

Simultaneously quantify surface protein & cytokine expression

Figure 3. Quantify variability in activation profile and cytokine release from PBMCs isolated from different donors.

SKOV-3 cells were seeded (4K/well) in a co-culture assay with PBMCs (10K/well) from 3 donors. Activation was induced by CD3/CD28 Dynabeads. Cells were lifted following the adherent cell lifting protocol after 4 days and analyzed using the T Cell Activation Cell and Cytokine Profiling Kit. (A-C) Expression of markers of early (CD69), late (CD25) and even later (HLA-DR) T cell activation. (D) IFNγ release by each donor.

Easy to use and interpret; pre-defined gating strategy

Figure 4. Pre-defined gates on Forecyt enable automatic phenotyping of human T cell subsets

Minimal sample manipulation: low volume, one wash, no dilution

  • Simple protocol where cells and supernatant are taken as one sample allowing for a streamlined workflow.

  • No labelling, or compensation optimization steps required.

Figure 5a. Easy to follow protocol for the analysis of T cell phenotypes and cytokine release using the T Cell Activation Cell and Cytokine Profiling Kit.

Figure 5b. Easy to follow modified protocol to allow the inclusion of target cells in the assay. Allowing the analysis of T cell phenotypes and cytokine release using the T Cell Activation Cell and Cytokine Profiling Kit.

Ordering Information

 

T Cell Activation Cell and Cytokine Profiling Kit 
PlatformCompatible with iQue3 VBR configuration
Available SizesCatalog NumbersOrder Now
1 x 96 well90560T Cell Activation Cell and Cytokine Profiling Kit
5 x 96 wells90561
1 x 384 wells90562
5 x 384 wells90563

 

Technical Resources

Documents

T Cell Activation Cell and Cytokine Profiling Kit Manual

 

Application Notes

A Kinase inhibitor phenotypic screen using a multiplex T cell activation assay

Combined Live Cell and Flow Cytometry Analysis of Immune Cell Killing

 

Posters

Routine monitoring of T cell activation by multiplexing analysis of immunophenotypes and secreted cytokines

Characterization of bi-specific T-cell engager (BiTE) antibody-mediated cell killing using a combined live-cell and flow cytometry workflow

Rapid, High Capacity Monitoring of T-Cell Activation for Adoptive Cell Therapy

Back to Top