Local and systemic adverse events (AEs) possibly, probably or definitely related to vaccination shown as percentage of volunteers affected.

<p>(a) Post ChAd63 CS 5×10⁹ vp; (b) Post ChAd63 CS 5×10¹⁰ vp; (c) Post MVA CS 2×10⁸ pfu.</p

Summary of PBMC IFN-y ELIspot responses of volunteers in each group.

<p>Summed SFC/million PBMCs. (A) and (B) individual responses for groups 1A and 1B respectively over time. (C) and (D) show individual responses for groups 2A and 2B respectively over time. (E) median ELIspot response by group by time-point, changes were significant over time; for group 1B, day 0 to 14 p = 0.0063; day 14 to 63 p = 0.0148; day 0 to 63 p = 0.0002, Mann-Whitney test; and group 2B, day 0 to 14 p = 0.01; day 14 to 63 p = 0.0074; day 0 to 63 p = 0.0002, Mann-Whitney test. (F), (G) & (H) show individual responses by group at days 14, 63 and 84 or 90 respectively.</p

CONSORT diagram of study progress.

<p>39 volunteers were screened. Reasons for not meeting the inclusion criteria in 7 excluded volunteers were: psychiatric morbidity (2), history of malignancy (2), one each of: history of headaches, Carbohydrate Deficient Transferrin (CDT) >3% and neutropenia.</p

Local, systemic and laboratory adverse events post immunization.

<p>Adverse events deemed possibly, probably or definitely related to vaccination are shown. ‘Other systemic’ following MVA CS included nasal congestion, laryngitis and pharyngitis. The highest intensity adverse event per subject is listed. Other local AEs included paraesthesia. All ‘other’ AEs were considered possibly related to vaccination due to a temporal association.</p><p>*The two laboratory AEs related to the same volunteer, who experienced neutropenia following both vaccination.</p><p>**Three severe systemic AEs followed MVA CS, all experienced by one volunteer simultaneously and resolved within 48 hours of vaccination.</p><p>Local, systemic and laboratory adverse events post immunization.</p

Multifunctional antigen specific cells.

<p>Percent of CD4⁺ and CD8⁺ PBMCs at day 63 producing 1–4 cytokines following stimulation with CS peptide pools.</p

Cytokine production by cell type and time-point assessed by 7 colour flow cytometry.

<p>Mean percent and standard error of the mean (SEM) of CD4+ and CD8+ PBMCs producing antigen-specific cytokines at given time-point post vaccination are shown for each cytokine. (A) percent CD4+ and (B) percent CD8+ PBMCs producing CD107a, IFNg, IL2 and TNFa at day 63. (C) percent CD4+ and (D) percent CD8+ PBMCs producing CD107a, IFNg, IL2 and TNFa at day 63. (E) percent CD4+ and (F) percent CD8+ PBMCs producing CD107a, IFNg, IL2 and TNFa at day 84.</p

Individual ELISpot responses in SFC/10⁶ PBMC at day 63 by peptide pool.

<p>Bar represents mean, whiskers; standard error of the mean.</p

Median & Interquartile range ELISpot responses in SFC/10⁶ PBMC at day 63 by peptide.

<p>Box plots of the medians and 25% and 75% percentiles of response to each peptide. The first and third quartiles are the top and base of each box, the upper and lower bars represent the maximum and minimum values respectively.</p

Assessment of antibody isotype profiles following vaccination and CHMI. Isotype profiles of serum antibody responses were assessed by ELISA.

<p>(A) Anti-MSP1₁₉ responses in the VAC037 Phase Ia clinical trial (open symbols) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107903#pone.0107903-Sheehy1" target="_blank">[19]</a> and the VAC039 Phase IIa clinical trial (closed symbols) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107903#pone.0107903-Sheehy3" target="_blank">[21]</a>. Responses are shown at baseline (d0); following ChAd63 MSP1 priming immunization (d28); following the MVA MSP1 boost (d84 in the Phase Ia trial or dC−1 in the Phase IIa trial); or following CHMI at day of diagnosis, dC+DoD, or at first follow-up post drug treatment, dC+35. (B) Anti-AMA1 responses in the VAC036 Phase Ia clinical trial <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107903#pone.0107903-Sheehy2" target="_blank">[20]</a> and VAC039 Phase IIa trial <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107903#pone.0107903-Sheehy3" target="_blank">[21]</a>, reported as in panel A. In all panels, individual and median responses are shown, n = 4<b>–</b>13 depending on sample availability for each tested time-point.</p

Associations between IgG antibody responses and CHMI outcome measures.

<p>Associations are reported between anti-MSP1₁₉ or anti-AMA1 total IgG ELISA titer readouts at various time-points and/or fold-change post-CHMI (as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0107903#pone-0107903-g001" target="_blank">Figure 1E</a>), as well as with time to malaria diagnosis by thick-film microscopy during CHMI, and parasitemia at time of diagnosis (measured by qPCR in terms of parasites/mL blood). In all cases, Spearman’s rank correlation coefficient and <i>P</i> value are shown. n/a = not applicable; n.d. = not done. *For these analyses, relevant vaccine groups were included: for the MSP1₁₉ analysis, data were combined from the MSP1-only vaccination group and from the MSP1+AMA1 and MSP1+ME-TRAP co-administration groups; and for the AMA1 analysis data were combined from the AMA1-only vaccination group and from the MSP1+AMA1 co-administration group.</p><p>Associations between IgG antibody responses and CHMI outcome measures.</p