Enzyme-Linked Immuno Spot (ELISpot) is a technique which quantifies immune cells of low abundance which release biomarkers such as cytokines in response to antigenic stimulation. ELISpot is a highly sensitive method to test immune modulators, novel vaccine candidates or de-risk immunogenicity testing in an antigen-specific CD4 and/or CD8 T cell assay.
Condition | Aim |
No Stimulation | Negative control |
PMA | Positive control |
CERI (CMV, EBV, RSV, Influenza) | MHC-I restricted peptide pool to evaluate modulation of CD8+ T-cell memory response |
CPI (CMV, Parainfluenza, Influenza) | Positive protein antigens to evaluate modulation of CD4+ T-cell memory response |
CEF (CMV, EBV, Influenza) | MHC-I restricted peptide pool to evaluate modulation of CD8+ T-cell memory response |
Cyclosporin A (CsA) | Inhibition of immune response |

Spot Forming Units (SPU) for IFN-γ per 100,000 PBMC from CERI, CPI and CEF antigens for three donors


Evaluation of therapeutic modulation of Th17 differentiation

Figure 1: Polarisation of Th17 cells. Naïve CD4 cells were cultured under Th17 polarising conditions for 12 days in the presence or absence of Ursolic Acid. CD4 T cells were assessed for proliferation by CTV dilution; intracellular cytokine staining (ICS) of IL-17 and IL-10 by flow cytometry. On Day 12, Supernatants were collected and evaluated for IL-17 and IL-10 levels by MSD. Ursolic Acid showed selective inhibition of IL-17 production by ICS and MSD.
Evaluation of therapeutic modulation of antigen-specific memory T cell responses to recall antigens
I. T cell response to Tetanus Toxoid, Influenza and PPD antigens

II. Dose response to Influenza antigen

Antigen specific T cells response to a recall antigens. (I) Healthy donors PBMC were stimulated with PHA-M or triple antigen cocktail (Tetanus Toxoid, Influenza and PPD). Cyclosporin was used as a reference treatment. (II) Dose response to Influenza antigen. CD4 and CD8 T cells proliferation was measured by flow cytometry using CTV dilutions.
Enzyme-Linked Immuno Spot (ELISpot) is a technique which quantifies immune cells of low abundance which release biomarkers such as cytokines in response to antigenic stimulation. ELISpot is a highly sensitive method to test immune modulators, novel vaccine candidates or de-risk immunogenicity testing in an antigen-specific CD4 and/or CD8 T cell assay.
Condition | Aim |
No Stimulation | Negative control |
PMA | Positive control |
CERI (CMV, EBV, RSV, Influenza) | MHC-I restricted peptide pool to evaluate modulation of CD8+ T-cell memory response |
CPI (CMV, Parainfluenza, Influenza) | Positive protein antigens to evaluate modulation of CD4+ T-cell memory response |
CEF (CMV, EBV, Influenza) | MHC-I restricted peptide pool to evaluate modulation of CD8+ T-cell memory response |
Cyclosporin A (CsA) | Inhibition of immune response |

Spot Forming Units (SPU) for IFN-γ per 100,000 PBMC from CERI, CPI and CEF antigens for three donors

Evaluation of therapeutic modulation of antigen-specific memory T cell responses to recall antigens
I. T cell response to Tetanus Toxoid, Influenza and PPD antigens

II. Dose response to Influenza antigen

Antigen specific T cells response to a recall antigens. (I) Healthy donors PBMC were stimulated with PHA-M or triple antigen cocktail (Tetanus Toxoid, Influenza and PPD). Cyclosporin was used as a reference treatment. (II) Dose response to Influenza antigen. CD4 and CD8 T cells proliferation was measured by flow cytometry using CTV dilutions.