Journal: Infection and Immunity

Article Title: Mucosal Vaccination against Tuberculosis Using Inert Bioparticles

doi: 10.1128/IAI.00786-13

Figure Lengend Snippet: IFN-γ responses. IFN-γ was determined from splenocytes stimulated using an ELISA or ELISPOT assay. (a and b) ELISA detection of IFN-γ from splenocytes stimulated with rMPT64 (a) or with rAcr or rAg85B (b). (c) ELISPOT assay determination of the number of spot-forming units (SFU) in splenocytes stimulated with rMPT64 or rAcr-Ag85B. Spleens were taken from mice (n = 2) immunized using dosing regimen 1. The BCG group received one dose of BCG.

Article Snippet: Free binding sites were blocked with 10% FCS in PBS at RT for 1 h. Culture supernatants were tested at RT for 2 h, and IFN-γ was detected with a biotinylated anti-mouse IFN-γ–streptavidin–horseradish peroxidase conjugate (BD Biosciences) at RT for 1 h. Recombinant mouse IFN-γ (BD Biosciences) was used as a standard.

Techniques: Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Journal: Infection and Immunity

Article Title: Mucosal Vaccination against Tuberculosis Using Inert Bioparticles

doi: 10.1128/IAI.00786-13

Figure Lengend Snippet: Gating strategy for polyfunctional T cell analysis. Representative FlowJo scatter plots of a naive mouse splenocyte sample are presented to show the gating strategy. Samples were first gated by forward scatter (FSC-A)/side scatter (SSC-A) to select lymphocytes and were then gated by FSC-H/FSC-A to gate for single cells. This population was then gated for CD3+ CD4+ or CD3+ CD8+ staining. These two populations were then analyzed for IFN-γ, IL-2, and TNF-α by staining.

Article Snippet: Free binding sites were blocked with 10% FCS in PBS at RT for 1 h. Culture supernatants were tested at RT for 2 h, and IFN-γ was detected with a biotinylated anti-mouse IFN-γ–streptavidin–horseradish peroxidase conjugate (BD Biosciences) at RT for 1 h. Recombinant mouse IFN-γ (BD Biosciences) was used as a standard.

Techniques: Staining