Virus detection in tissues from pigs infected with <i>Zaire ebolavirus</i> (ZEBOV).

<p>Virus detection for pigs infected with <i>Zaire ebolavirus</i> in experiment 2 was done by quantitative real time RT-PCR targeting the L gene of ZEBOV and L gene copy numbers estimated based on a standard curve. Data represents log₁₀ copies/mL of tissue suspension (numbers in brackets represent standard deviations of 3 replicates for each sample). SLN, submandibular lymph node; BLN, bronchial lymph node; MLN, mesenteric lymph nodes, U, virus was either absent or below levels detectable by the current assay.</p

Induction of proapoptotic molecules in lungs from pigs infected with <i>Zaire ebolavirus.</i>

<p>Data was obtained by microarray analysis of RNA from lungs of pigs infected with <i>Zaire ebolavirus</i> in experiment 1. Numbers represent fold change in transcription relative to tissues from uninfected controls. DPI, days post infection. N = 2 pigs for controls, DPI 3, 5 and 7.</p

Up-regulation of pattern recognition receptors, transcription factors and interferon stimulated genes in lungs from pigs infected with <i>Zaire ebolavirus.</i>

<p>Data was obtained by microarray analysis of RNA from lungs of pigs infected with <i>Zaire ebolavirus</i> in experiment 1. Numbers represent fold change in transcription relative to tissues from uninfected controls. DPI, days post infection. N = 2 pigs for controls, DPI 3, 5 and 7.</p

IgM antibody response in <i>Zaire ebolavirus</i> (ZEBOV)-infected pigs.

<p>IgM was detected by a porcine IgM capture ELISA using serum from pigs in experiment 2. Data for individual animals is presented as adjusted OD₄₀₅ obtained by subtracting the OD₄₀₅ of a negative antigen from that of the ZEBOV antigen for 1/100 dilution of each serum sample. The horizontal broken line represents the cut-off OD₄₀₅ value defined as mean +3 standard deviation of negative control sera.</p

Systemic cytokine response in <i>Zaire ebolavirus</i> (ZEBOV)-infected pigs.

<p>(A) Serum IFN-α, and (B), serum IL-6 response were measured by ELISA. Data points represent means ± standard deviation for n = 6 pigs on 0, 1, 3 and 5 days post infection (DPI) and n = 3 pigs on DPI 6 from experiment 2. Histograms represent means ± standard deviation of fold change in mRNA transcripts for (C) IL-12; (D) TNF-α, and (E) IFN-γ in PBMCs (n = 6 pigs) from experiment 2.</p

Histopathology and immunohistochemistry findings in <i>Zaire ebolavirus</i> (ZEBOV) -infected pig lungs.

<p>(A). Hematoxylin and eosin stained slide showing infiltration of inflammatory cells into affected lobules and some fluid-filled alveoli. Bar = 200 µm. (B) The majority of the inflammatory cells within the lungs showed positive immunostaining with antibody Mac387 against L1 antigen. Bar = 200 µm. (C) Higher magnification of (B) showing several neutrophils (arrows) and a macrophage (arrowheads). Bar = 10 µm. (D) Double immunolabelling detected the simultaneous expression of Mac387 macrophage marker (pink) and ZEBOV antigen (brown, arrows) within the same cell indicating the presence of viral antigen within macrophages. Viral antigen was not detected within neutrophils (arrowhead). Bar = 10 µm. (E) Numerous multinucleated syncytial cells (arrow) were observed within alveolar spaces. H&E stain. Bar = 10 µm. <i>Inset:</i> Serial section of lung with specific immunolabelling for ZEBOV antigen in multinucleated syncytial cells (arrows) and mononuclear inflammatory cells (arrowhead). (F) CD3 positive T-lymphocytes were limited to infiltrates in the bronchiolar lamina propria and peribronchial areas (arrows). <i>Inset:</i> Higher magnification of perbronchiolar CD3 positive T-lymphocytes. Lung sections were obtained from pigs in experiment 2.</p

Chemokine and acute phase response in lungs from pigs infected with <i>Zaire ebolavirus.</i>

<p>Data was obtained by microarray analysis of RNA from lungs of pigs infected with <i>Zaire ebolavirus</i> in experiment 1. Numbers represent fold change in transcription relative to tissues from uninfected controls. DPI, days post infection. N = 2 pigs for controls, DPI 3, 5 and 7.</p

Changes in PBMC subsets in <i>Zaire ebolavirus</i> (ZEBOV)-infected pigs.

<p>(A) CD172a+ (monocytes/macrophages and dendritic) cells (B) CD14− CD172a+ (dendritic) cells, (C) CD21+ B lymphocytes and (D) CD3+ T lymphocytes in PBMCs. Data points for (A–D) represent means ± standard deviation for n = 6 pigs on 0, 1, 3 and 5 days post infection (DPI) and n = 3 pigs on DPI 6 from experiment 2. (E) a representative plot showing infection of porcine monocytes with ZEBOV determined by flow cytometry. Filled histogram represents the uninfected cells and the open histogram for ZEBOV-infected monocytes for n = 3 different PBMC donor pigs.</p

Model for pulmonary disease pathogenesis in <i>Zaire ebolavirus</i> (ZEBOV)-infected pigs based on microarray data.

<p>Arrows show the proposed sequence of events that take place from the time of ZEBOV infection to the development of lung inflammation and respiratory distress. The grey shaded blocks represent the anti-viral/anti-inflammatory machinery. The scale suggests that the series of events favour the proinflammatory pathway.</p

Plasma levels of VSVΔG/ZEBOVGP from rhesus macaques after vaccination.

<p>Plasma levels of VSVΔG/ZEBOVGP from rhesus macaques after vaccination.</p