Lymphocyte Proliferation assay
The characterization of T cells is crucial for understanding their role in disease and immunotherapeutic interventions such as vaccinations or tolerizing therapies. In a proliferation assay a complete protein or parts of proteins (peptides) can be used to stimulate CD4+ and CD8+ T cells. This assay provides useful information about the presence of antigen specific T cells and gives indications of T cell immunogenicity and can therefore be of aid to drug safety assessments.
Antigen specific proliferation of T and B cells can be measured by flow cytometry. CFSE labelled cells lose half of their staining after each division. Original % of antigen specific cells can be calculated be determining the % of cells in each division cycle (as shown on the right).
From the supernatant of the PBMC cultures cytokine production can be measured to provide information on the type of response after antigen stimulation.
As a result of the SEB stimulation 33% of the cells went into cell division resulting in 75% dividing cells after 9 days.
Unstained cells are shown in gray and CFSE labelled cells are shown in green.
- CD40L assay
- CD8: MHC multimer analyses (combinatorial coding)
- Immunogenicity analyses
- Therapy efficacy
- Therapy safety
- Mechanism of Action
Boks MA, Kager-Groenland JR, Mousset CM, van Ham SM, ten Brinke A.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. Clin Immunol. 2014; 151(2):136-45.
Boks MA, Zwaginga JJ, van Ham SM, ten Brinke A. An optimized CFSE-based T-cell suppression assay to evaluate the suppressive capacity of regulatory T-cells induced by human tolerogenic dendritic cells. Scand J Immunol 2010; 72(2):158-168.