iQue® T Cell Activation Cell and Cytokine Profiling Kit by Sartorius Group

The activation of naive T cells by antigen is one of the central events in the adaptive immune responses.  Binding of an antigen to the T cell receptor (TCR)/CD3 complex along with costimulatory signals initiates clonal expansion of both CD4+ helper and CD8+ cytotoxic T cells. 

In addition to cell proliferation, a variety of signaling pathways are activated leading to the expression of functional proteins including cell surface markers and the release of cytokines. Further differentiation of activated T cells result in effector cell functions such as B-lymphocyte and macrophage activation by CD4+ cells and T cell mediated cell killing by CD8+ cells. 

Understanding the biology of T cell activation as well as profiling biologics and compounds that impact T cell activation are critical for a broad spectrum of applications including drug discovery and adoptive cell therapy. 

• Speed: Sample to actionable data by allowing multiple assays to be run in one assay reducing the hands on time, numbers of overall tests and conserving  precious sample. 
• Results: 15 endpoints from a single well assay measuring T cell phenotypes, early and late cell activation markers in each phenotype, cell viability, cell count, proliferation and secreted effector cytokines. 
• Biological insight: Generate cell health and functional data from same cell by multiplexing cell and cytokine detection in one well. 
• Simplicity: Single platform assay and data acquisition eliminates the time of acquiring and correlating data from multiple sources. 

The T Cell Activation Cell and Cytokine Profiling Kit assesses the activation status of T cells while providing information about their health and their role in cytokine secretion. This one wash assay requires minimal hands on time and measures both cells and beads to provide the following biological insights: 

• Live immune cells are distinguished from dead cells by staining with a fluorescent membrane integrity dye. 
• Cells are immunophenotyped by staining with a fluorescent antibody panel to separate CD3+ T cells, CD3- non-T cells, CD4+ T helper cells, CD8+ T cytotoxic cells. 
• The T cell phenotypes are profiled for the expression of 3 activation markers , CD69 (early), CD25 (late), and HLA-DR (even later, with strong stimulation). 
• Optimized workflow enables the measurement of effector cytokines secreted by activated T cells , Th1 cytokine IFNγ/ multifunctional cytokine TNFα in the same assay well."