IL-12p70 blocking assay

Protocol: IL-12p70 blocking assay

In order to assess the importance of IL-12p70 in balancing Th1/Th17 response two different experiments are performed.
In one case, DCs are stimulated for 8 hours with live spores of S. cerevisiae and C. albicans hyphae at a stimuli:DC ratio of 4:1 in the presence of different concentration of human recombinant IL12p70 (0, 1, 10 and 100 ng/ml).
In the second experiment, DCs are pre-incubated with different concentrations (0, 0.1, 1, 10, 100 microg/ml) of a monoclonal anti-human IL-12 antibody for 2 hours, then stimulated for 8 hours with live S. cerevisiae cells in a stimuli:DC ratio of 4:1.
The ability of these stimulated DCs to drive CD4+ T cells polarization towards a Th1 response is assessed by measuring of IFNgamma and IL-17 in T cells 5-day culture supernatants.

created over 14 years ago (21 January 2010)    last modified over 13 years ago (28 September 2011)   [ RDF ]   [ RelFinder ]