Cytokine analysis - ELISA / CBA

Cytokine production can be measured with different methods. With a cytokine Enzyme-Linked Immuno Sorbent Assay (ELISA), the concentration of ex vivo cytokine levels in serum and plasma, induced cytokine production in our whole blood assays or cytokines produced in culture medium is measured.

This method can be used to screen for the immunogenicity and immunomodulatory properties of compounds and biologicals with regards to innate inflammatory responses, antigen-specific responses, CD4⁺ T cell skewing and many more immune reactions.

Advantages of cytokine production analysis by ELISA

• Cost-effective
• High-throughput
• Monitoring of cumulative cytokine production throughout the culture period

Performing ELISAs is an efficient method to test many samples in different dilutions in one experiment. If, however, you are more interested in measuring various cytokines simultaneously in a small sample volume, the Cytometric Bead Array (CBA) may be more suitable. The CBA is flow cytometry-based, which conveniently allows multiplexing. Alternatively, we also offer secreted protein detection via Luminex for detection of up to 500 different proteins.

Example of cytokine secretion detection by ELISA

Effect of differently induced tolerogenic dendritic cells (DCs) on CD4⁺ T cell activation

Different types of tolerogenic DCs (matured with indicated compounds) and monocyte-derived DCs (moDCs) were cultured at a 1:10 ratio with allogeneic naive CD4⁺ T cells. IL-10, IL-13 and IFN-γ production was determined by ELISA of the culture supernatant at day 6. The cultures were performed in duplicate and mean ± SEM of 5 independent experiments is shown.

Assessing the effect of a compound on a broad panel of cytokines produced by immune cells via a whole blood assay

PBMCs isolated from healthy donor whole blood (n=3) was cultured together with potentially immunostimulatory compounds for 6 hours. Serum was collected and cytokine production was assessed by CBA. Graphs show triplicate results from 1 representative donor.

Reference

IL-10-generated tolerogenic dendritic cells are optimal for functional regulatory T cell induction-a comparative study of human clinical-applicable DC
Boks MA, Kager-Groenland JR, Haasjes MS, Zwaginga JJ, van Ham SM, ten Brinke A 
Clin Immunol. 2012 Mar;142(3):332-42. doi: 10.1016/j.clim.2011.11.011.