Figure 6
QS-21 elicited innate cell recruitment and CD4/CD8 T-cell responses are dependent on Caspase-1.
(A) Bone-marrow derived dendritic cells (BMDCs) from mice lacking NALP3 or Caspase-1 expression were primed overnight with MPL and stimulated with either QS-21 or alum. IL-1β release in the supernatant was determined by ELISA. The data is represented as mean and SEM of 3 independent experiments. (B) WT or caspase-1 KO mice were injected i.m. with either PBS or QS-21 and the draining lymph nodes were recovered at the indicated time points. Whole lymph nodes were lysed and caspase-1 and IL-1β processing were detected by western blot. (C) Innate cell recruitment to the iliac DLN in WT and Caspase-1 KO mice 24 h post QS-21 injection. Absolute numbers of monocytes (Lin-CD11b+ Ly6C+ Ly6G−), neutrophils (SSChi Lin− CD11b+ Ly6Cint Ly6G+) and dendritic cells (Lin− CD11c+ MHCII+) per DLN were assessed by flow cytometry (PBS: n = 5, QS-21: n = 5–10). (D–F) Mice were immunised as in 1 A. At day 21, cytokine production (D) and polyfunctionality (E) of HBs-specific CD4 were evaluated by flow cytometry. (F) Cytokine production of HBs-specific and frequency of OVA-specifc CD8 T cells were evaluated by flow cytometry (Ag: n = 3, QS-21: n = 9–10). (G) Anti-HBs IgG1 and IgG2c titres in the serum at day 21 were measured by ELISA (Ag: n = 5–6, QS-21: n = 20). Each point represents one mouse and the horizontal bar represents the geometric mean. Statistical significance was determined by a non-parametric Mann-Whitney test. The data is representative of 2 independent experiments.
