Fig. 3

IFNγ is required for the induction of adaptive immune responses after immunization with an AS01-adjuvanted vaccine. a Transcriptional analysis was performed as in Fig. 2. DEGs at 4 h post immunization in the “Inflammation interferon_signalling” process network (MetaCore) are represented in a heatmaps showing their −log2FC over PBS and their interplay classification. b, c C57BL/6 mice were immunized once with HBs + indicated adjuvants (AS01 = MPL + QS-21 MPL = 100 µg/ml, QS-21 = 100 µg/ml, 2 × 10 µl injections, i.m.). dLNs were isolated at indicated time points and homogenized. b Cytokine concentration as assessed by Luminex (n = 5 mice/group, 1 out of 2 independent experiments is shown). c Levels of IFNγ mRNA transcript in the dLN as measured by RT-PCR (n = 5 mice/group). d Timeline of immunization protocol. Mice were immunized with AS01 and HBs/OVA (HBs = 80 µg/ml; OVA = 20 µg/ml; AS01 = 100 µg/ml MPL + 100 µg/ml QS-21, 2 × 50 µl/injection i.m.) twice at a 2 week interval. At day 21 levels of antigen-specific antibodies were titrated in the serum. Spleens were collected and splenocytes were restimulated with a pool of peptide spanning the length of HBs or OVA and analyzed by ICS. e–g C57BL/6 or Ifngr^−/− were immunized as in c. N = 6/condition, one out of two independent experiment is shown. e Frequency of total cytokine-producing antigen-specific CD4⁺ T cell. f Frequency of antigen-specific CD4⁺ cytokine-producing species. g Frequency of OVA-specific SIINFEKL-H2Kb⁺ CD8⁺ T cells. h Levels of HBs-specific IgG and ratio of HBs-specific IgG1/IgG2c in the serum. i–k WT mice were treated with an αIFNγ mAb (XMG1.2, 100 µg/mouse i.p.) and 48 h later immunized with AS01 and HBs/OVA as in d. i Frequency of total cytokine-producing antigen-specific CD4⁺ T cells. j Frequency of antigen-specific CD4⁺ T cells by cytokine expression. k Frequency of OVA-specific SIINFEKL-H2Kb⁺ CD8⁺ T cells. l HBs-specific IgG measured by ELISA in the serum. (N = 5/condition, one representative out of two independent experiments is shown. All graphs show mean ± SEM