SmRho loading efficiency on chitosan particles before and after coating with alginate.

<p>SmRho loading efficiency on chitosan particles before and after coating with alginate.</p

Protective effect and liver granuloma size induced by C57BL/6 mice vaccination and challenged with 25 <i>S. mansoni</i> cercariae.

*<p> <b>Statistically significant compared with the control group (p<0.05).</b></p

Release profile of protein SmRho from alginate-coated chitosan nanoparticles in SGF (A) and SIF (B) at 37°C (mean ± SD, n = 3).

<p>Release profile of protein SmRho from alginate-coated chitosan nanoparticles in SGF (A) and SIF (B) at 37°C (mean ± SD, n = 3).</p

Reactivity of sera from control and <i>S. mansoni</i> infected patient, drug treated or untreated, against SmRho and SWAP observed by ELISA^# (A) and against SmRho, SWAP, SEA and schistosomula extract by Western Blotting (B).

<p>M: Standard molecular weight Page Ruler Unstained Protein Ladder (Fermentas), 1: SWAP; 2: SEA, 3: schistosomula antigens; 4: recombinant protein Rho1-GTPase de <i>S. mansoni</i>. ^#The results are presented as the mean absorbance measured at 450 nm. Statistically significant differences of sera from infected patient with the control group, non infected patient, in each sera dilution evaluated, are indicated by (*) for p<0.05. The molecular weight markers, from top to bottom, are: 116, 66, 45, 35, 25, and 18 kDa.</p

Size and zeta potential of chitosan particles before and after coating.

<p>Size and zeta potential of chitosan particles before and after coating.</p

Cytokine profiles of mice immunized with coated SmRho-chitosan nanoparticles.

<p>Splenocytes isolated from mice immunized with CH-Rho-Alg, CH-Rho-Cpg-Alg, and CH-Rho-Alg (i.m.) were assayed for IL-10 (A) and INF-γ (B) production in response to <i>in vitro</i> stimulation with SWAP (25 µg/ml), rSmRho (25 µg/ml), SEA (25 µg/ml) or medium alone as control. <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0001894#s3" target="_blank">Results</a> represent the mean ± SD of each group. *Statistically significant differences between cytokines produced after SWAP, rSmRho or SEA stimulation compared with unstimulated splenocytes (control) (p<0.05).</p

Treatment groups and the formulation administered to each one.

*<p>NPs: Nanoparticles.</p

Expression and purification of rSmRho.

<p>(A) Western Blotting profile of the <i>E. coli</i> (BL21 pRARE) transformed with the pDEST42-SmRho. Lane: MW – Prestained protein molecular weight marker (Fermentas). Lanes 1 and 2 represent a clone before and after induction with 0.5 mM IPTG, respectively. Lanes 3 and 4: soluble and insoluble fraction after <i>E. coli</i> (BL21 pRARE) lysis, respectively. (B) SDS-PAGE 12% profile of Ni²⁺ chromatography. Lanes: MW – Unstained protein molecular weight marker (Fermentas); 1 - flow through; 2–5 fractions of rSmRho–6×HIS-tag fusion protein eluted after Ni2+ chromatography. Positions of molecular mass standards (kDa) are indicated. Arrows indicate the purified rSmRho.</p

Oligonucleotides design to amplify the cDNA clone coding SmRho sequence.

<p>The <i>attB</i> sites, required for cloning in Gateway System, are underlined.</p