Introduction
While there is no industry-accepted protocol for measuring the functionality of peripheral blood mononuclear cells (PBMC), it’s an important test that should be conducted for quality control.
We needed a reliable, reproducible way to measure the functionality of our cryopreserved PBMC, so we developed a custom assay using recall antigens to understand the in vitro activity of our cells. By testing PBMC for immune response to several different antigens over time, we intended to better understand an acceptable range of variation in recall response from PBMC samples from the same donor over time.
The recall antigen testing assay needed to effectively evaluate the quality of important immune cell types, including T cells, B cells, and monocytes. We strategically designed the assay to use several antigens:
- Phytohaemagglutinin (PHA): As a positive control
- Lipopolysaccharide (LPS): To stimulate B cells and monocytes
- Cytomegalovirus (CMV): As a natural, chronic infection for a subset of the CMV-infected population
- Tetanus Toxoid: Due to its widespread use in vaccinations