5,000 NAU/kg of α-ANDV FFP is sufficient to protect hamsters from lethal HPS disease.

<p>A) Survival curve of hamsters challenged with 4,000 PFU of ANDV i.n. on day 0 and passively transferred with dilutions of α-ANDV FFP on day 8. P-values were determined by comparing FFP dilution to no antibody control. B) α-N ELISA endpoint titers (log₁₀) were conducted with sera from surviving hamsters challenged with ANDV in A). GMT for each group are shown.</p

α-ANDV FFP effectively neutralizes ANDV <i>in vitro</i> and is detectable in hamsters after passive transfer.

<p>A) Neutralizing antibody titers were determined by ANDV PRNT₈₀ performed on α-ANDV FFP and normal human serum. * indicates results are statistically significant. B) Neutralizing antibody bioavailability was determined by ANDV PRNT₈₀ performed on hamster serum samples collected after passive transfer of α-ANDV FFP (64,000 NAU/kg) by either s.c. or i.m. route (3 hamsters per group) on day 0, through 21 days. PRNT₈₀ titers represent the lowest serum dilution neutralizing 80% of the plaques relative to the control (no serum). PRNT₅₀ values of a single group are denoted by a dashed line.</p

12,000 NAU/kg of α-ANDV FFP and α-ANDV duck IgY/IgYΔFc protects hamsters from lethal HPS disease.

<p>A) and B) Survival curve of hamsters that were challenged with 4,000 PFU i.n. of ANDV on day 0 and passively transferred with α-ANDV FFP or α-ANDV duck IgY/IgYΔFc on day 5 postinfection (A) or day 8 postinfection (B). * indicates statistical significance when compared to normal IgY/IgYΔFc treatment. C) α-N ELISA endpoint titers (log₁₀) were conducted with sera from surviving hamsters challenged with ANDV in A) and B). GMT for each group are shown. * indicates results are statistically significant when compared to no antibody controls.</p

Neutralizing activity of α-ANDV duck IgY/IgYΔFc <i>in vitro</i> and <i>in vivo</i>.

<p>A) Western blot analysis of IgY components recognized by α-duck IgY antibodies. A) represents IgY/IgYΔFc by Western blot with SDS-PAGE run under non-reducing conditions and probed with an α-duck IgY antibody recognizing the heavy chain of both IgY and IgYΔFc. HC is the heavy chain of IgY. B) Percent neutralization of α-ANDV FFP and α-ANDV duck IgYΔFc measured by ANDV PRNT. Dotted line represents 80% neutralization (PRNT). C) Neutralizing antibody bioavailability was determined by ANDV PRNT performed on hamster serum samples collected after passive transfer of α-ANDV duck IgYΔFc (12,000 NAU/kg and 64,000 NAU/kg) by the s.c. route on day 0, through 21 days. PRNT₈₀ titers (solid lines) and PRNT₅₀ titers (dashed lines) are plotted. D) Survival curve of hamsters challenged with 4,000 PFU of ANDV i.n. on day 0 and passively transferred with 5,000 NAU/kg of α-ANDV duck IgYΔFc on day 8 postinfection.</p

12,000 NAU/kg of anti-ANDV duck IgY/IgYΔFc protects hamsters from lethal HPS disease.

<p>A) Survival curve of hamsters that were challenged with 200 PFU i.m. of ANDV on day 0 and passively transferred with α-ANDV FFP (8 hamsters per group) or α-ANDV duck IgY/IgYΔFc (16 hamsters per group) on day 5 postinfection. * indicates statistical significance when compared to normal IgY/IgYΔFc treatment. B) α-N ELISA endpoint titers (log₁₀) were conducted with sera from surviving hamsters challenged with ANDV in A). GMT for each group are shown.</p

α-ANDV FFP passively transferred on days 5 and 8 protects hamsters from lethal disease and infection.

<p>A) Survival curve of hamsters challenged with 4,000 PFU i.n. of ANDV on day 0, then passively transferred with α-ANDV FFP (30,720 NAU/kg) on days 5, 8, 12 or 15 postinfection. Rabbit sera (administered at 1,920 NAU/kg) were collected 102 days post DNA vaccination <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035996#pone.0035996-Hooper2" target="_blank">[16]</a>. P-values were determined based on comparison to normal serum on matching day. B) α-N ELISA endpoint titers (log₁₀) were conducted with sera from surviving hamsters challenged with ANDV in A). GMT for each group are shown. * indicates results are statistically significant when compared to rabbit sera positive control.</p

Levels of c8A and c7D11monoclonal antibodies in infected and uninfected marmosets.

<p>Marmosets were subcutaneously injected with a 1:1 mixture of a cocktail containing c8A and c7D1 (20mg/Kg of each antibody per animal). A single animal was not exposed to monkeypox virus (#11). Of the exposed animals, Days 1, 12 and 21 were analyzed for the presence of c7D11 (A) and c8A (B). For infected animals #8, #9 and #10, days are relative to the day of exposure. For animal #11, days are relative to subcutaneous injection of the antibody.</p

Comparison of the clinical course of smallpox and monkeypox in humans and infected nonhuman primates.

<p>To elucidate both the resemblance to human smallpox disease and how our model relates to the “gold standard” of NHP models for poxvirus countermeasures, the figure depicts the human disease (grey outlined box), the IV monkeypox-macaque model (red outlined box) and the IN monkeypox-marmoset model (outlined in green). Arrows are color coded by host and indicate the point of exposure. Days (in the form of a number line) relative to exposure are given for each Human disease manifestations are temporally identical to IN exposed marmosets to include viremia and oral shedding (not shown-see <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0006581#sec015" target="_blank">discussion</a>), but fever has not yet been detected in marmosets (IV = intravenous; IN = intranasal). This Figure is a modification of Figure 34 from Mucker, 2014 [<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0006581#pntd.0006581.ref036" target="_blank">36</a>].</p

Skin of a mock treated marmoset exhibiting a classical smallpox rash.

<p>Previous to exposure with 1000 PFU of monkeypox virus, animal 5 was treated with non-pox-virus specific antibody. This animal manifested a typical smallpox rash as can be seen on the exposed (hairless) portions of the abdomen and groin (A) a zoomed in portion of the abdomen is shown in B.</p

Viremia and viral shedding in marmosets exposed to intranasal monkeypox virus.

<p>Whole blood and oral swabs were processed and titrated onto BSC-1 cells. Two of four animals had detectable levels of virus in blood samples on the Day 9. Virus from oral swabs was detectable as early as day 6 (high dosed animal #4). Limits of detection for whole blood (dashed orange line) and for oral swabs (dashed blue line). All animals were positive to varying extents. Notice the onset of oral shedding relative to viremia.</p