Intracellular Cytokine Staining

Our flow cytometry assays provide intricate and detailed information about the immune responses by combing intracellular cytokine staining with cell surface markers for immunophenotyping and MHC multimer staining for antigen-specific responses. 

cytokine profiling

Intracellular cytokine staining for flow cytometric readout is the basis of multiple assays we develop to characterize the immune response by multicolor flow cytometry. It can be combined with cell surface markers for immunophenotyping such as maturation markers and proliferation markers.  

At ImmuMap, we routinely use intracellular cytokine staining to provide a detailed analysis of the cytokine-secreting T cells in samples from clinical human patient samples, preclinical animal samples or cell cultures (cryopreserved from clients or grown in our own culturing unit), often combined with staining of activation markers such as CD69.

Activated cell populations for intracellular cytokine staining can be prepared from in vivo samples from clinical human patients, preclinical animal models or in vitro cell cultures using a wide range of cell stimulants such as non-specific agents, compounds from our clients, short or long peptides or whole proteins. We either test cell samples provided by client from human or mouse in vivo studies or utilize our in-house cell bank for ex vivo testing of client compounds.

Importantly, although we are focusing on T cells, the T cell stimulators and the cytokines secreted from different T cell subsets, we are also capable of developing intracellular cytokine staining assays dealing with other cell populations, e.g. Natural Killer (NK) or dendritic cells.

Applications of intracellular cytokine staining by flow cytometry:

Detection, enumeration and isolation of phenotypic and functional antigen-specific T cells or other cells responding to the stimulant in question in a single sample.

Determination of the exact phenotype of responding cell populations with up to 18 different customizable parameters.

Confirmation and further characterization of cytokine secretion by antigen-specific cells as a follow up on previous ELISPOT data.

The main advantage of intracellular cytokine staining compared to these methods is the possibility of simultaneously analysing numerous parameters, including the cytokine production alongside multiple phenotypic, differentiation and functional markers that can be found both intracellularly and extracellularly. Most notably, the detection of multiple cytokines produced by responding T cells is commonly assessed using intracellular cytokine staining and flow cytometry. Thus, several cytokines, e.g. IFNg, TNFa and IL-2, can be detected alongside differentiation markers, e.g. CD3, CD4 and CD8, in a single experiment.

For an even more accurate determination of antigen-specific cells, intracellular cytokine staining can be used in conjunction with other functional assays such as MHC multimer staining or ELISPOT

We offer intracellular cytokine staining analyses for human, murine and porcine samples.

Learn more about
Common MHC Multimer Staining

Learn how ImmuMap utilizes our specialty within MHC multimer staining to detect antigen-specific T cell responses. MHC multimer staining is used heavily for investigating epitope spreading and tracking immunotherapy induced immune responses.