The Correlation of IL-10 expression with FoxP3 and TGFβ1 mRNA Expression.

<p>For the A) baseline (r = 0.557 <i>P</i> = 0.030), b) 7 days following vaccination (r = 0.695 P = 0.019) and C) 28 days following vaccination time points (r = 0.554 <i>P</i> = 0.048) IL-10mRNA expression correlates with FoxP3 expression. At the 28 days time point D) IL-10 mRNA also correlates with TGF-β1 mRNA expression (r = 0.642 <i>P</i> = 0.023). The volunteers with sterile protection are indicated by open circles and triangles indicate volunteers who did not enter the challenge study. Correlations were performed using Spearman's one-sided test, n = 9–12.</p

Foldchange of Gene Expression at Each Timepoint in CS-Stimulated Cells Compared to Unstimulated Cells.

<p>Day 0 =  baseline prior to vaccination.</p><p>Vac+7 =  seven days after the final vaccination.</p><p>Vac+28 = 28 days after the final vaccination.</p

Inverse Correlation between Anti-inflammatory Cytokines and Antibody Response on the Day of Malaria Challenge.

<p>The anti-CS IgG antibody response on the day of malaria challenge induced by vaccination with RTS,S and MVA-CS inversely correlates with the anti-inflammatory cytokines TGF-β1 and IL-10 measured in unstimulated PBMC. A) TGF-β1 at baseline inversely correlates with anti-CS IgG measured on day of challenge r = −0.644 <i>P</i> = 0.022. B) TGF-β1 at day 7 inversely correlates with anti-CS IgG measured on day of challenge, r = −0.670 <i>P</i> = 0.009. C) IL-10 mRNA at baseline inversely correlates with anti-CS IgG measured on day of challenge r  = −0.554 <i>P</i> = 0.031. D) IL-10 mRNA at day 28 inversely correlates with anti-CS IgG measured on day of challenge r = −0.762 <i>P</i> = 0.005. The volunteers with sterile protection are indicated by open circles and triangles indicate volunteers who did not enter the challenge study. Correlations were performed using Spearman's two-tailed test, n = 9–12.</p

The Expression of Cytokines in CS-stimulated Cells Before and After Vaccination.

<p>Expression of IFN-γ, MIG, TGF-β1, FoxP3 and IL-10 was measured by real-time RT-PCR in total PBMCs following 12 hour culture with the vaccine antigen CS in subjects who received vaccination with RTS,S/AS02A and MVA-CS. Results are expressed as copy number relative to the housekeeping gene HPRT. Median values (9 subjects) are shown. Vac+7 =  seven days after the final vaccination, Vac+28 = 28 days after the final vaccination, each subject received two doses of RTS,S/AS02A and one dose of MVA-CS. A) IFN-g mRNA, B) MIG mRNA, C) TGF-β1 mRNA, D) FoxP3 mRNA, E) IL-10 mRNA and F) Correlation of MIG mRNA expression at day 7 with days to parasitemia following sporozoite challenge, n = 6–12.</p

The Expression of Cytokines in Unstimulated Cells Before and After Vaccination.

<p>Expression of IFN-γ, MIG, TGF-β1, FoxP3 and IL-10 was measured by real-time RT-PCR in total PBMCs following 12 hour culture in media only in subjects who received vaccination with RTS,S/AS02A and MVA-CS. Results are expressed as copy number relative to the housekeeping gene HPRT. Median values (6–12 subjects) are shown. Vac+7 =  seven days after the final vaccination, Vac+28 = 28 days after the final vaccination, each subject received two doses of RTS,S/AS02A and one dose of MVA-CS. A) IFN-g mRNA, B) MIG mRNA, C) TGF-β1 mRNA, D) FoxP3 mRNA, E) IL-10 mRNA and F) Correlation of MIG mRNA expression at day 7 with days to parasitemia following sporozoite challenge.</p

TRAP peptide specific IFN-γ and FoxP3 mRNA expression increase in the FFM ME-TRAP vaccine group over time.

<p>A) IFN-γ mRNA expression B) FoxP3 mRNA expression C) IL10 mRNA expression D) TGF-β1 mRNA expression (n = 10–20).</p

Correlation of qPCR with the IFN-γ ELISPOT assay is dependent on the frequency of antigen specific T cells present and the quantity of viable cells tested.

<p>A) The qPCR detection of IFN-γ secreted by a known number's of mouse CD8 T cell clone's. The detection limit is >25 and <250 T cell clones/10⁶ splenocytes. B) Spearman's correlation of TRAP peptide stimulated IFN-γ measured by qPCR and ELISPOT improves as the cut-off for HPRT copy number increases. The dashed line indicates a p value of 0.05.</p

Description of subjects selected for qPCR study.

<p>Description of subjects selected for qPCR study.</p

Correlation between IFN-γ mRNA 1 week following vaccination and baseline IL10, FoxP3 and TGF-β1 mRNA.

<p>There was no correlation between antigen specific IFN-γ and A) IFN-γ at screening B) IL10 at screening C) FoxP3 at screening D) TGFB at screening (n = 20).</p

Mean expression of the different cytokines/markers at different time points (log copy number/HPRT).

<p>The mean mRNA expression levels were compared between the two vaccination groups.</p