Developmental stage of the spleens of adult SCID and NMRI nude mice.

<p>125 to 135 days old SCID and NMRI nude mice from different providers were imaged with MRI to evaluate the developmental stage of the spleens (arrows). For SCID mice, animals purchased from Charles River (A) and Harlan (B) showed poorly developed, diffuse, and small spleens while animals from Taconic (C) had solid, compact, and relatively large spleens. NMRI nude mice from Charles River (D) and Scanbur (E) showed fully developed spleens with a similar appearance.</p

Visualization of primary tumor in the spleen and liver metastasis.

<p>Laparotomy observations of the entire organs and sections of the organs where the tumors are engrafted give an accurate idea of the location and shape of the spleen and liver tumors within the organ, which corresponds to its predicted location and shape by MRI.</p

Development of intrasplenic and intrahepatic tumors in SCID mice.

<p>Four human colorectal cancer cell lines (Co115, HCT116, SW620, HT29) were tested for their ability to induce growth of intrasplenic and intrahepatic tumors in SCID mice. 1x10e6 cells were injected into the spleen and primary tumor growth (A), number of metastasis in the liver (B), and total volume of the liver tumors (C) were analyzed weekly by MRI. Each line represents an individual animal. Number of animals used for testing each cell line is presented in parentheses.</p

Morphology of the spleens of 30- and 125-day-old SCID and NMRI nude mice.

<p>SCID, severe combined immunodeficiency; NA, not applicable.</p

Validation of HT29 cell line in the metastatic colorectal cancer model.

<p>1x10e6 cells were injected into the spleen of 125-day-old SCID mice and intrasplenic tumor growth (A), number of metastatic lesions in the liver (B), and total volume of liver metastases (C) were assessed weekly with MRI. Each dot represents an individual animal and mean of each time point is marked with dotted line. Number of animals analyzed at each time point is presented in parentheses.</p

MRI based evaluation of the effect of oncolytic adenovirus Ad5-D24-RGD in metastatic colorectal cancer mouse model.

<p>1x10e6 HT29 cells were injected into the spleen of 125-day-old SCID mice. Intratumoral injection of the virus (OV) at a dose of 2x10e7 viral particles was given 21 days later (arrow). Intrasplenic tumor growth (A), number of metastatic lesions in the liver (B) and a total volume of liver metastasis (C) were assessed weekly with MRI. Data is presented as mean ± SD. *, p<0.05, **, p<0.01.</p

Immunohistochemical analysis of Ad5 and Ad5T122 infection in ex vivo tissue cultures of normal human liver and colorectal carcinoma liver metastasis.

<p>Precision-cut tumour (top and third row of panels) and liver (second row and bottom panels) tissue cultures were left uninfected (left panels) or infected with 10⁷ PFU (in 2 ml of media) of Ad5 (middle panels) or Ad5T122 (right panels), and fixed five days later for staining with haematoxylin-eosin (panels G-L) or an antibody against the viral E1A protein (panels A–F). Necrosis developing in the central area of the uninfected control tissue (panel J) is indicated with arrows.</p

Regulation of Cox-2 and VEGF promoter activity with anti-inflammatory reagents.

<p>Monolayers were preincubated with reagents, and C33A (a, b), SiHa (c, d), Caski (e, f) and HeLa (g, h) cells were infected with 1 000 viral particles (vp)/cell of Ad5luc1 (with the CMV controlling luciferase), Adcox-2Mluc or AdVEGFluc, and luciferase expression was analyzed. Transgene expression level with Cox-2 and VEGF promoters are compared to the CMV promoter (%). Each point represents the mean of four experiments±standard error. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.0001 <i>versus</i> no substance.</p

Oncolytic adenoviruses display efficient killing of cervical cancer cells <i>in vitro</i> and <i>in vivo</i>.

<p>(a–d) Monolayers were infected with RGDCRADcox-2R, Ad5/3VEGF-E1, Ad5-Δ24RGD, wild-type adenovirus and Ad5luc1 (<i>E1</i>-deleted control virus). Cell viability was measured with MTS assay. The OD₄₉₀ values of uninfected cells were set as 100%. Data is expressed as mean±standard error of quadruplicate experiments. (e–f) C33A cells were injected subcutaneously into nude mice and advanced tumors were allowed to develop. The mice were treated either with (e) three intratumoral injections of 1×10⁹ viral particles (vp) of Ad5luc1, wild-type virus or oncolytic adenoviruses on three consecutive days, or with (f) a single intravenous injection of 1×10¹¹ vp. *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.0001 <i>versus</i> Ad5luc1. Bars indicate standard error.</p

Comparison of cell killing by Ad5 and Ad5T122 in a panel of cancer cell lines.

<p>Four cell lines of non-hepatic (HCT116, A549, and Hep-2) or hepatic (Huh7) origin were infected with Ad5Luc1 (non-replicative virus control), Ad5, or Ad5T122 at an MOI of 0.05. Cell survival in the infected cells was measured 7 days (Hep-2 and A549) or 9 days (HCT116 and Huh7) post-infection using an ATP-based cell viability assay, and plotted on the y-axis as the percentage of the control values measured from uninfected cultures. The data are presented as the mean of six repetitions ± standard error.</p