Teton™ CytoProfiling, T Cell Activation (Jurkat Cells)

Background

T cell activation is a central event in adaptive immunity, orchestrating responses against pathogens and tumors while also shaping transplant rejection and autoimmunity. To capture the dynamic process of activation, Jurkat T cells were stimulated with anti-CD3 and anti-CD28 antibodies and profiled across three time points (30 minutes, 6 hours, and 24 hours), alongside positive and negative controls. Learn more about this study in our data spotlight.

Methods

Using the Teton Immuno Panel Kit and three 24-plex Teton Focus Panels (T Cell Activation, Cytokine Signaling, and Innate Immunity), we simultaneously measured 343 RNA, 122 protein and phospho-protein, and 6 cell paint markers across thousands of single cells with spatial resolution. Protocol details can be found in the Teton CytoProfiling User Guide.

Results

Multiomic profiling revealed a clear temporal progression of T cell activation. Early responses (30 min) included rapid induction of immediate-early genes such as NR4A1 and surface marker CD69, followed by metabolic ramping and costimulatory signaling (e.g., CD28, IL2RA) at 6 hours. By 24 hours, Jurkat cells displayed sustained activation signatures, including CD84 upregulation and high TOX expression, suggesting transition toward an exhaustion phenotype. Phospho-protein analysis further confirmed signaling activity, with phospho-RPS6 elevated at early time points. This integrated single-cell dataset highlights the power of Teton CytoProfiling to resolve dynamic immune activation trajectories with multiomic and morphological depth .