Proliferation assay - flow cytometry

Measuring lymphocyte proliferation upon stimulation can help you determine the frequency of T cells responsive to an antigen of interest, and it can give insight into modulation of this response by immunomodulatory compounds. Whether you are investigating effects on allogeneic reactions, immunogenicity or antigen-specific reactions, analyzing proliferation by flow cytometry will answer your questions. 

If you want to know more about the functionality of T cells in patients that respond to your treatment, you can combine the proliferation assay with assays in which we assess their cytokine profile or transcription factor expression. 

Our lymphocyte proliferation assays are based on labelling of the cells with a fluorescent proliferation dye. This allows tracking of each cell division, because it will result in a decrease in fluorescence. This loss of fluorescence can be quantified for each cell type specifically, using flow cytometry. This way, the number of cell divisions in response to stimulation can be determined. 

Proliferation assay- flow cytometry

 

Also see proliferation assay by 3H thymidine incorporation.

Example

Proliferation assay

Violet stained PBMCs from a healthy donor have been stimulated with either CMV or TT. After 8 days, both antigens were able to elicit a T cell response, which could be measured by analyzing the violet intensity. When analyzing transcription factor expression of the proliferating cells, a clear difference is visible between the CMV and TT stimulated cells. The type of memory T cells that responds can be analyzed by transcription factor expression or cytokine production, together with the proliferation assay.

Contact

Need help?

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