Rapid, no-wash measurement of immune checkpoint molecules and cytokines in co-cultures of immune cells and cancer cell lines

Breast cancer tumors can adapt to immune cell infiltration by responding to the increased concentration of interferon gamma (IFN-ɣ) and other cytokines secreted by subsets of T lymphocytes with the upregulation of the immune checkpoint proteins such as Programmed cell death ligand 1 (PD-L1).

These checkpoint proteins allow the tumors to evade immune targeting and reduce the immune response, thus promoting tumor progression.

In this application note, you will learn:

• How exogenous addition of IFN-ɣ effects the expression of PD-L1 and secretion of several cytokines in cultures of HCC38 cells (a triple-negative breast cancer cell line)
• How co-culturing activated immune cells and breast cancer cells stimulates differential expression of some immune checkpoint and inflammatory biomarkers compared to culturing cells alone with PBMC-conditioned media.
• How to rapidly measure multiple biomarkers in cell culture supernatant and lysates from the same wells of a culture dish to examine protein expression profiles from cancer and immune cell culture models using AlphaLISA assays together with ATPlite 1step and the EnVision multimode plate reader.

For research use only. Not for use in diagnostic procedures.