Abortive vaccinia virus replication and maturation in SCCF1 cells.

<p><b>A)</b> At 48 hours after infection, the replication centre (viral factory) is inconspicuous, only a few immature viruses (IV) were found around it. <b>B)</b> Two IVs were visible, one IV showed nucleoid inside thus can be identified as IVN. Neither mature viruses nor enveloped viruses were detected throughout all infected cells. <b>C)</b> there are two small immature viruses (IV) located within cell and in the edge of an abnormal granular virosome (VS) region. No MVs were visible there. <b>D)</b> The immature viruses (IV) migrate to peripheral of the cell, underneath cell membrane. They still fail to get membranous envelope even at this late stage of nearly being released out.</p

In vitro cytotoxicity and and transduction of vvdd in SCCF1 cells.

<p><b>A)</b> SCCF1 cells were infected with vvdd with 0.01, 0.1 or 1 pfu/cell. Viability of cells was measured on day 3. <b>B)</b> Compared to A549 cells, SCCF1 cells maintained their viability significantly better on day 3 <b>C)</b> When SCCF1 cells formed a tight monolayer before infection, the cells were still alive 10 days after infection. <b>D)</b> SCCF1 cell were infected with 0.04, 0.2, 1 or 5 pfu per cell and luciferase expression was measured 4 h postinfection in relative light units.</p

Electron microscopy of negative stained, purified vaccinia viral particles released from cultured cells.

<p>Supernatant from infected SCCF1 or A549 cells was collected, purified and negative staining electron microscopy was performed. <b>A-C:</b> viral particles produced by A549 cells: mature particles of brick-shaped, with outer envelope (OE) derived from cell membrane. <b>D-E</b>: immature vaccinia viral particles produced by SCCF1 cells: the particles are irregular in shape and in electron density. Their internal contents are not tightly packed, no clear boundary or membranous outer envelope is visible.</p

Portions of final results in days and treatment groups.

<p>ALQ–samples with detection of adenoviral sequence above limit of quantification (positive samples), BLQ–below limit of quantification, BLD–below limit of detection, N.D.–not detected.</p

Production of virus in SCC-F1 cells.

<p><b>A)</b> Infected SCC-F1 cells constantly produce infectious particles to the supernatant. <b>B)</b> Virus released to the supernatant by infected SCCF1 cells was able to re-infect both A549 and SCC-F1 cells. <b>C)</b> Viral particles released to the supernatant were completely neutralized with anti-VV antibody.</p

Vaccinia virus replication and maturation in A549 cells.

<p><b>A-B</b>: After enter A549 cells, at 48 hours post infection, vaccinia viral particles begin their normal life cycle quickly: form replication centre (viral factory) in juxta-nuclear region manifested as a homogeneous, low-to-middle electron density, void of any organelles. The replication begins with formation of crescent (Cr), then immature virus (IV), immature virus with nucleoids (IVN), mature virus (MV). Many MV are wrapped by intracellular membrane from Trans-Golgi cistern, form intracellular enveloped virus (IEV). IEVs move towards cell periphery, fuss with cell membrane and obtained another envelope from cell membrane, finally they are released to extracellular space to form extracellular enveloped virus (EEV). Some EEV remain attached on cell membrane as cell associated extracellular enveloped virus (CEV). <b>C-D</b> Different intermediate particles from cells other than cells in a-b in high magnification.</p

Portions of final results in sample types from D2-Bio group in days 3, 29 and 183.

<p>ALQ–samples with detection of adenoviral sequence above limit of quantification (positive samples), BLQ–below limit of quantification, BLD–below limit of detection, N.D.–not detected.</p

Portions of final results in samples of serum from TOX groups in day 29, 190 and 260.

<p>ALQ–samples with detection of adenoviral sequence above limit of quantification (positive samples), BLQ–below limit of quantification, BLD–below limit of detection, N.D.–not detected.</p

Body weight (g)–TOX groups.

<p>(A)–males. (B)–females.</p

Food consumption–TOX groups.

<p>(A)–males. (B)–females.</p