Principle of Immune Cell Activation Assay

It involves exposing responder immune cells (e.g., PBMCs, T cells, B cells, NK cells) to specific stimuli, such as chemical compounds, antigens, or cytokines.
Stimulating agents, like antigens or cytokines, engage receptors on immune cells, leading to signaling cascades.
Activated immune cells upregulate activation markers (e.g., CD69, CD25) and secrete cytokines (e.g., IFN-γ, IL-2).

Model Description

Allogeneic stimulator cells (e.g., PBMCs, dendritic cells) are co-cultured with responder T cells, leading to TCR-mediated activation. This interaction triggers signaling pathways that induce T cell activation, proliferation, and cytokine secretion.

Readouts

T cell activation by flow cytometry and T cell response by cytokine secretion (IFN-γ, IL-2, and TNF-α release).

Add-On Services

Evaluation of cytokine release, proteomics, and transcriptomics.

Key Components of Immune Cell Activation Assay

1. Stimulator Cells

PBMCs which serve as both the responder and stimulator cells, or antigen-presenting cells (e.g., dendritic cells).

2. Immune Modulators

a. Activators: Agents that stimulate immune cells, such as cytokines (e.g., IL-2, IL-12), pathogen-associated molecular patterns (PAMPs), or synthetic activators (e.g., PMA, ionomycin).

b. Inhibitors: Agents that block immune cell activation or modulate immune responses (e.g., checkpoint inhibitors like anti-PD-1, anti-CTLA-4 antibodies, or immunosuppressive cytokines like TGF-β).

3. Activation Markers

Monoclonal antibodies targeting cell surface activation markers (e.g., CD69, CD25, CD137) or intracellular markers to assess immune cell activation.

4. Detection Methods

a. Flow Cytometry: For measuring activation markers, cytokine production, and cellular proliferation.

b. Cytokine Assays: ELISA or multiplex assays to quantify the levels of secreted cytokines.