Protection and prepatent period after heterologous NF135.C10 challenge.

<p><b>Left panels:</b> Kaplan-Meier curves showing percentage of thick smear negative volunteers after NF135.C10 challenge according to previous NF54 protection status (A) and NF54 CPS-immunization dose (C). <b>Right panels:</b> The corresponding distribution of prepatent period of thick smear positive volunteers is shown in dot plots according to NF54 protection status (B) and NF54 CPS-immunization dose (D). Lines represent medians. ⨂ = Volunteer presumptively treated after NF54 challenge and considered NF54 protected.</p

Study flow diagram.

<p>The previous NF54 CPS-immunization study is shown in grey. P = NF54 protected, NP = NF54 unprotected. ⨂ = Volunteer presumptively treated on day 10.5 after NF54 challenge and considered NF54 protected</p

Adverse events before and after initiation of treatment.

<p>Average number of possibly and probably related (both solicited and unsolicited) AE per previously NF54 protected or control volunteer in relation to the time of positive thick smear (day of treatment). Time points are plotted towards day of treatment, depicted as ‘T’, from 3 days before until 7 days after start of treatment.</p

Parasitemia before and after treatment.

<p>Parasitemia measured by qPCR up until initiation of treatment (<b>A</b> and <b>C</b>) and from treatment onwards (<b>B</b> and <b>D</b>) in previously NF54 protected volunteers <b>(A</b> and <b>B)</b> and controls <b>(C</b> and <b>D)</b>. Each line represents an individual subject with the same colour before and after treatment. Values shown as 25 Pf/ml were negative (i.e. half the detection limit of the qPCR: 50 parasites/ml).</p

Study flow diagram.

<p>Thirty-six subjects were screened for eligibility, of whom twenty were included in the trial and randomized over three groups. One control subject was excluded after initiation of chemoprophylaxis but before the first immunization because of an unexpected visit to a malaria-endemic area during the study period. In a double-blind fashion, fifteen subjects received either CPS-CQ or CPS-MQ immunization and four control subjects received bites from uninfected mosquitoes and mefloquine prophylaxis. Subjects received a challenge infection by bites of five infected mosquitoes sixteen weeks after discontinuation of prophylaxis.</p

Antibody responses induced by CPS-CQ and CPS-MQ immunization.

<p>Antibodies against CSP (A and B; in AU), LSA-1 (C and D), and MSP-1 (E and F) were analyzed at baseline (B), 28 days after the first (I1) and second (I2) immunization and one day before challenge (C-1; 20 weeks after the last immunization) for all CPS-CQ (A, C and E, n = 5) and CPS-MQ (B, D and F, n = 10) immunized volunteers. Data are shown as individual titers with medians. Open squares indicate protected subjects, filled circles indicate unprotected subjects. Differences between the time points were analyzed by Friedman test with Dunn’s multiple comparison post-hoc test. Significant differences are indicated by asterices with * (p<0.05), ** (p<0.01), *** (p<0.001).</p

Additional file 2: Figure S1. of Comparative assessment of An. gambiae and An. stephensi mosquitoes to determine transmission-reducing activity of antibodies against P. falciparum sexual stage antigens

The ranking of estimates of TBA of antibodies against Pfs48/45 (mAb 85RF45.1 and mAb 85RF45.5) and Pfs25 (mAb 32F81 and mAb 4B7) in An. gambiae and An. stephensi mosquitoes. TBA of transmission effective mAb 85RF45.1 (blue), mAb 85RF45.5 (red), mAb 32F81 (green) and mAb 4B7 (black) in An. gambiae depending on TBA in An. stephensi mosquitoes. Dots and triangles represent the predicted TBA, while lines represent 95% confidence intervals in An. gambiae and An. stephensi. (TIFF 1302 kb

Parasitemia during CPS immunization.

<p>Parasitemia was determined retrospectively, once daily from day 6 until day 10 after each immunization, by real-time quantitative PCR (qPCR). Each line represents an individual subject from the CPS-MQ (dashed blue lines) or CPS-CQ group (red lines). The number of subjects with a positive qPCR/total number of volunteers in the CPS-MQ (blue) and CPS-CQ (red) groups after each immunization are shown above the graph. Values shown as 17.5 on the log-scale were negative (i.e. half the detection limit of the qPCR: 35 parasites/ml).</p

Additional file 1: Table S1. of Comparative assessment of An. gambiae and An. stephensi mosquitoes to determine transmission-reducing activity of antibodies against P. falciparum sexual stage antigens

TRA and TBA estimates of human serum IgG samples. IgG concentration is shown in mg/ml. TRA estimates are shown as % reduction in oocyst intensity with 95% CI and TBA estimates are shown as % reduction in oocyst prevalence with 95% CI. (XLSX 31 kb

Adverse events during CPS immunization.

<p>Percentage of volunteers in each group experiencing possibly or probably related AE after the first (I), second (II) and third (III) immunization. AEs were evaluated at each visit and graded for severity as described in the methods paragraph: mild (light grey), moderate (dark grey) and severe (black). Only the highest intensity per subject is listed. No Serious Adverse Events occurred.</p