T Cell Functional Assays

  • ELISpot: Measures the frequency of IFN-g-secreting T cells in peripheral blood mononuclear cells (PBMC). Is more sensitive than intracellular cytokine staining (ICS).
  • ICS: Measures the frequency of antigen-specific T cells secreting numerous cytokines/chemokines (e.g. IFN-g, TNF-a, IL-2). Also determines phenotype of responding T cells (CD4 or CD8).
  • Sorting of HIV-infected and/or TB-infected cells
  • MHC Restriction: Determines which HLA allele is used to restrict a specific T cell response.
  • Viral Suppression Assay (VSA): Measures the ability of HIV-specific T cells to inhibit viral replication in autologous CD4 cells. Can be performed with bulk T cells or with T cell lines/clones specific for certain HIV epitopes.
  • Cytotoxicity Assays: Measures the direct killing activity of HIV-specific T cells. This is a flow-based assay.
  • Proliferation Assays: Measures ability of T cells (both CD4 and CD8) to proliferate in response to HIV antigens/epitopes. This is a flow-based assay using CFSE.
  • Treg Suppression Assay: Measures the suppressive effects of Tregs on HIV-specific effector cells (both CD4 and CD8). Can measure suppressive effects of Tregs on either cytokine secretion or proliferation of effectors.
  • siRNA transfection of primary T cells (including Tregs).


For more information on how to access these services, contact Ana Gervassi at ana.gervassi at seattlebiomed.org.