Cytokine analyses- intracellular cytokine staining

Intracellular cytokine staining (ICS) is a method of cytokine analyses that will provide information on the subtype of T helper cell that is induced. Rather than the amount of secreted cytokines produced by cells in culture, which is generally analyzed by ELISA or CBA, ICS will give you information on the proportion of cells producing certain cytokines.

This approach is especially suited to combine with the following assays:

This can help, for example, to unravel the polarization, strength and specificity of immune responses upon immune-modulating therapies.

Example: 

Anti-TNFα polarizes T cells towards IL-10 and away from IFNγ

DC- T cell MLR- cytokines by intracellular cytokine staining

 

 

Naïve CD4+ T cells were primed by tolDC for 2 weeks, subsequently restimulated with PMA and ionomycin and stained intracellularly for IL-10. A representative dotplot of 14 independent experiments is shown. Percentage of IL-10, IFNγ and IL-4 positive T cells determined by intracellular staining of restimulated primed T cells.
Mean + SEM of 14. *p≤0.05, **p≤0.01, ***p≤0.001.

Reference

Inhibition of TNF receptor signalling by anti-TNFα biologicals primes naïve CD4+ T cells towards IL-10+ T cells with a regulatory phenotype and function. 
Boks MA, Kager-Groenland JR, Mousset CM, van Ham SM, ten Brinke A. Clin Immunol 2014;151(2):136-45

Contact

Need help?

Get in touch with Anja ten Brinke, PhD or Annelies Turksma, PhD
Immunomonitoring@Sanquin.nl