SipD protein identity sequences for the 4 major <i>Salmonella</i> pathogens of humans.

<p>SipD protein identity sequences for the 4 major <i>Salmonella</i> pathogens of humans.</p

Summary of the antibody responses (IgG and IgA) after the last immunization with PrgI or SipD by the SC, IN and OG routes.

<p>Summary of the antibody responses (IgG and IgA) after the last immunization with PrgI or SipD by the SC, IN and OG routes.</p

Summary of the antibody responses (IgG and IgA) after the last immunization with both proteins (PrgI and SipD) by the SC, IN and OG routes.

<p>Summary of the antibody responses (IgG and IgA) after the last immunization with both proteins (PrgI and SipD) by the SC, IN and OG routes.</p

Serum Ig(G+M) concentrations of mice immunized with PrgI or SipD (A) and PrgI/SipD (B).

<p>Serum Ig(G+M) antibodies specific for PrgI (left) and SipD (right) were quantified by sandwich ELISA 2 weeks after the last immunization as described in Materials and Methods. Data represent mean concentrations (ng/mL) and the standard errors (SEM) from 14–16 individual mice per group. Asterisks *** indicate <i>P</i> value< 0.001, comparing the antibody responses using different routes versus control mice. No cross-reactions were observed between PrgI and SipD (data not shown). [°: indicates injected immunogen; *: indicates biotinylated recombinant protein].</p

IgG (2a +2b) / IgG 1 ratio after PrgI (left) and SipD (right) immunizations.

<p>Mice immunized with PrgI or SipD separately are represented on panel A and those receiving both PrgI and SipD on panel B. Data represent mean and the standard errors (SEM) from 14–16 mice per group. [°: indicates immunogen injected; *: indicates biotinylated recombinant protein].</p

IgA titers of mice immunized with PrgI or SipD (A) or PrgI/SipD (B).

<p>Serum IgA antibodies specific for PrgI (left) and SipD (right) were measured by sandwich ELISA 2 weeks after the last immunization as described in Materials and Methods. Data represent mean titers and the standard errors (SEM) from 14–16 individual mice per group. Asterisks indicate <i>P</i> values: *** <i>p</i> < 0.001, ** 0.001<<i>p <</i>0.01 and * <i>p</i>< 0.05 when comparing mice immunized by the IN or OG route versus mice immunized by the SC route and control mice. No cross-reactions were observed between PrgI and SipD (data not shown). [°: indicates injected immunogen; *: indicates biotinylated recombinant protein].</p

Protection efficacy of PrgI and SipD T3SS proteins in mice from lethal challenge with <i>S</i>. Typhimurium.

<p>Protection efficacy of PrgI and SipD T3SS proteins in mice from lethal challenge with <i>S</i>. Typhimurium.</p

Production of cTA12 in <i>Sf9</i> cells by infection of a recombinant baculovirus at different multiplicities of infection (MOI).

<p>1.35×10⁷ cells seeded in T75 flasks were infected at different MOI (•: 0.04; ▪: 0.12; <b>▴</b>0.4; ⧫1.2) with the recombinant baculovirus stock. cTA12 containing supernatants were harvested at different times and antibody was quantified by enzyme immunoassay.</p

Sensitivity of the optimized Pla45/Pla35* immunoassay.

<p><i>Y. pestis</i> CO92 was grown at 37°C (dark blue), 28°C (medium blue), or 20°C (light blue), and subjected to the optimized sandwich ELISA. The graphs represent the lowest AU values. The limit of detection (LoD) is represented by a horizontal black line. Vertical lines represent the standard deviation of duplicates.</p

Specificity, sensitivity and comprehensiveness of the PLA-dipstick.

<p>Each strain was cultured at its optimal growth temperature (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0054947#s2" target="_blank">Materials and methods</a>). The bacterial suspensions were adjusted to ca. 10⁹ cfu/ml and serially diluted. 100 µl of these suspensions were used for the dipstick assays.</p><p>NA: not applicable.</p><p><b>+</b>: positive test line and/or <i>Y. pestis</i> aggregates line.</p><p><b>−</b>: negative test line and <i>Y. pestis</i> aggregates line.</p