Protection in passively immunized mice following i.n. challenge with <i>B. pseudomallei</i> strain 1026b.

<p>BALB/c mice were administered 1 mg of either CPS IgG3 mAb 3C5 or LPS IgG3 mAb 4C7 alone or 1 mg of each mAb in combination by the i.p. route. Intranasal challenge was performed 18 h later with 15 LD₅₀ of <i>B. pseudomallei</i>. Mice were monitored for 21 days after which gross pathology and spleen cfu were determined on survivors (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035386#pone-0035386-t001" target="_blank">Table 1</a>). Control mice were treated with 1 mg of an irrelevant IgG3 mAb. <i>p</i> values of survival vs. controls are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035386#pone-0035386-t001" target="_blank">Table 1</a>.</p

Detection of CPS within a splenic abscess by IHC.

<p>Organs were harvested from control BALB/c mice (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035386#pone-0035386-g003" target="_blank">Fig. 3</a>) that were infected with <i>B. pseudomallei</i> strain 1026b. A tissue section from a spleen that contained multiple large abscesses is shown (left panel). Location of CPS was identified by HRP-labeled mAb 3C5 (brown). Box within the panel on the left indicates the boundary of an abscess and surrounding normal splenic tissue (tissue within box is magnified in right panel). White scale bars indicate 50 µm.</p

Protection in passively immunized mice following i.n. challenge with <i>B. pseudomallei</i> strain K96243.

<p>mAbs were administered by the i.p. route at the doses (µg) listed. Intranasal challenge was performed 18 h later with 2 LD₅₀ of <i>B. pseudomallei</i>. Mice were monitored for 21 days. Control mice were treated with 1 mg of an irrelevant IgG3 mAb. <i>p</i> values of survival vs. controls are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035386#pone-0035386-t001" target="_blank">Table 1</a>.</p

Survival and gross pathology of mice passively treated with mAbs.

a<p><i>p</i> value vs. controls determined from Kaplan-Meier survival plots by log-rank (Mantel-Cox) test, bold values are statistically significant (<i>p</i><0.05).</p>b<p>positive spleen cfu was determined on survivors and assumed to occur in mice that died before study endpoint.</p>c<p><i>p</i> values vs. controls determined by Fisher's exact test, bold values are statistically significant (<i>p</i><0.05).</p>d<p>spleen cfu was assessed on survivors only; values indicate cfu determined by plating 100 µl from a 1 ml spleen homogenate; T indicates too numerous to count.</p>e<p>determination of abscess formation on internal organs was performed on survivors only.</p

Effect of mAb dose and combination therapy in mice challenged with <i>B. pseudomallei</i> strain 1026b.

<p>Mice were administered mAb(s) by the i.p. route followed 18 h later by i.n. challenge with 15 LD₅₀ of <i>B. pseudomallei</i>. (A) Dose-response experiment in which mice were treated with the doses (µg) listed of each mAb alone. (B) Multiple doses of mAbs 3C5 and 4C7 were administered in combination at the doses (µg) listed. Mice were monitored for 42 days after which gross pathology and spleen cfu were determined on survivors (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035386#pone-0035386-t001" target="_blank">Table 1</a>). Control mice were not treated with mAb. <i>p</i> values of survival vs. controls are listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035386#pone-0035386-t001" target="_blank">Table 1</a>.</p

Acai PS enhances the clearance of type A <i>F. tularensis</i> from RAW264.7 cells, but not murine BMDMs via NO.

<p>Cells were treated with Acai PS 16 hr prior to infection <i>F. tularensis</i> SchuS4 (MOI∼30), some wells were also pre-treated with 400 µM L-NMA.</p>a<p>Log₁₀ CFU/well from three wells/treatment shown; standard deviation in parentheses; results are representative of two independent experiments.</p>*<p>p<0.05 as compared to the same cell type not treated with Acai PS at the same time point.</p>∧<p>p<0.05 as compared to the same cell type, with the same Acai treatment, treated with L-NMA at 20 hr post-infection. ND = Not done.</p

Nasal administration of Acai PS confers protection against pulmonary <i>B. pseudomallei</i> infection.

<p>Female C57BL/6 mice (n = 5/group) were treated i.n. with PBS or with 100–1000 µg of Acai PS one day prior to, or immediately after, intranasal infection with 3×10³ CFUs of <i>B. pseudomallei</i> 1026b. A) Body weights and B) clinical scores were recorded daily, and C) on day 3, CFU determinations were performed in the lungs, spleens, and livers. Error bars depict SEM. *P<0.05 as compared to PBS group. **** indicates that *P<0.05 for all Acai PS-treated groups in relation to PBS-treated group at this time point. Data depicted in A–B) are representative of two independent experiments. The dashed line in C) indicates the limit of bacterial CFU detection.</p

Acai PS enhances LVS clearance from in human primary macrophages and enhances NK cell IFN-γ mRNA.

<p>Human primary macrophages (1×10⁴cells/well, 3 wells/treatment) were derived from PBMCS and infected with LVS (MOI∼300). One day prior to macrophage infection, autologous NK cells were also isolated via magnetic sorting. Macrophages and NK cells were treated separately with varying amounts of Acai PS 16 h prior to macrophage infection. After infection of the macrophages, fresh media with or without Acai PS or fresh media containing NK cells (∼20 NK cells/macrophage) with or without Acai PS were then added to the macrophage containing wells. A) Twenty h after infection, NK cells (non-adherent) were removed, macrophages were lysed, and intracellular bacteria enumerated. Error bars represent standard error. *P<0.05, as compared to untreated macrophages. B) Total RNA was extracted from NK cells co-cultured with LVS-infected macrophages (with or without Acai PS), and RT-PCR was performed for β-actin (control) and IFN-γ, TNF-α, IL-17A, IL-21, granzyme B, perforin, and TRAIL. Results are representative of independent experiments from five different blood donors.</p

Acai PS enhances IFN-γ by innate leukocytes during pulmonary type A <i>F. tularensis</i> infection.

<p>C57BL/6 mice (n = 5/group) were treated i.n. with 1 mg of Acai PS one day prior to i.n. infection with 50 CFUs of <i>F. tularensis</i> SchuS4. Some mice were also depleted of IFN-γ two days prior to infection. A) Intracellular expression of IFN-γ was determined for lung NK cells by flow cytometry, and two days after infection, and B) lung and splenic bacterial burdens were determined. Data are representative of two independent experiments. Error bars depict SEM. *P<0.05 as compared to PBS-treated animals receiving the same antibody treatment.</p

Acai PS augments IFN-γ responses by γδ T cells and NK cells during pulmonary infection.

<p>C57BL/6 mice (n = 5/group) were treated by the i.n. route with 1 mg of Acai PS one day prior to i.n. infection with 5×10⁵ CFUs of <i>B. thailandensis</i> E264. One day after infection, cells were harvested from lungs, and intracellular IFN-γ expression by γδ T cells and NK cells was determined by flow cytometry. Error bars depict SEM. *P<0.05 as compared to the same cell type from PBS-treated animals.</p