Article Figures & Data
Figures
- Fig. 1.
A, plasma mouse VEGF concentrations (24 hours) after escalating single DC101 doses (0–2,000 μg/mouse; five mice per group; age of mice, 6–8 weeks) in non–tumor-bearing SCID mice in undepleted and immunodepleted pooled samples (from five plasma samples). B, Western blot analysis of undepleted and depleted plasma samples showing the presence and absence of the DC101 antibody, respectively; membranes were blotted with horseradish peroxidase-conjugated goat antirat IgG antibody. +, immunodepleted plasma samples; −, undepleted samples. C, dose-dependent growth inhibition of HT-29 colorectal tumor xenografts after administration of escalating doses of DC101 every 3 days (five mice per group; age of mice, 6–8 weeks). Symbols and bars, mean ± SD.
- Fig. 2.
A, plasma mouse VEGF concentrations (24 hours) after single RAFL-1 escalating doses in non–tumor-bearing SCID mice. Symbols and bars, mean ± SD; five mice per group; age of mice, 6–8 weeks; P < 0.05, treated versus untreated control group, one-way analysis of variance, followed by the Student-Newman-Keuls test; two replicates per sample. B, in vitro human VEGF secretion by human PC-3 prostate and MDA-MB-231 breast cancer cell lines into conditioned medium. The results were the mean of two different experiments and were normalized for 106 cells (two replicates per sample). C, plasma human VEGF to tumor volume ratios after escalating single DC101 doses (0–1,500 μg/mouse) in MDA-MB-231 tumor-bearing mice after 24 hours. Symbols and bars, mean ± SD; five mice per group; age of mice, 6–8 weeks; P < 0.05, treated versus untreated control group, one-way analysis of variance, followed by the Student-Newman-Keuls test; two replicates per sample.
- Fig. 3.
A, plasma mouse VEGF kinetics during and after a three-injection DC101 schedule in non–tumor-bearing SCID mice (five mice per group; age of mice, 6–8 weeks) in immunodepleted and undepleted pooled samples (from five plasma samples); controls samples were at the limit of detection of the ELISA kit. B, plasma mouse VEGF concentrations after three-injection MF-1 schedule. Symbols and bars, mean ± SD; arrows, drug injections; three mice per group; age of mice, 6–8 weeks; ∗, P < 0.05, treated versus untreated control group of the same day, Student’s t test analysis; two replicates per sample. C, plasma mouse VEGF levels after three-injection RAFL-1 treatment during 21 days of observation. Symbols and bars, mean ± SD; arrows, drug injections; five mice per group; age of mice, 6–8 weeks; ∗, P < 0.05, treated versus untreated control group of the same day, Student’s t test analysis; two replicates per sample. D, comparative study between SCID and NIH Swiss nude mice plasma mouse VEGF levels in immunodepleted and undepleted pooled (from five mice) plasma samples after 28 days of simultaneous injections of DC101 (800 μg/mouse) and rat IgG (800 μg/mouse; control) every 3 days (five mice per group; age of mice, 6–8 weeks). Columns, mean value of double measurements of pooled samples.
- Fig. 4.
A, plasma human VEGF concentrations in MDA-MB-231 orthotopically xenotranplanted SCID mice, detected after 14 to 48 days after initiation of DC101 treatment alone or in combination with various metronomic low-dose chemotherapy treatments. DC101 was administered every 3 days, at 800 μg/mouse. Columns and bars, mean ± SD; ∗, concentrations are not detectable; five mice per group; age of mice, 6–8 weeks; ○, P < 0.05, treated versus untreated control group at time 0, unpaired Student’s t test analysis; two replicates per sample. Plasma human VEGF concentrations in an analogous experiment involving PC-3 human tumor xenotransplanted SCID mice were not detectable 20 to 51 days after initiation of treatment with DC101 alone or in combination with various metronomic chemotherapy treatments. B, DC101 (800 μg/mouse given on an every-3-day schedule, alone or in combination with low-dose metronomic chemotherapy regimens) inhibits the growth of orthotopic MDA-MB-231 tumors. Symbols and bars, mean ± SD; arrows, beginning of treatments. C, body weight of MDA-MB-231 tumor-bearing control mice and mice treated with DC101 alone or in combination with low-dose chemotherapy. No changes or decline in body weight were noted. Symbols and bars, mean ± SD. D, linear regression analysis between the duration of the experiment (expressed in days) per single treatment group and the mean of the latest available VEGF measurement of the same group. The mice were sacrificed when the mean tumor volume reached approximately 1,700 mm3. At least three points (control, VBL, and PTX groups) are superimposed.
- Fig. 5.
A, Northern blotting analysis of mouse VEGF and VEGFR-2 mRNA in lung, heart, and kidney tissues from SCID mice that were treated with DC101 or saline alone every 3 days. Total RNA was extracted, and approximately 20 μg were loaded per lane. Probes were labeled using [32P]dCTP. β-Actin mRNA is shown as loading control. Grouped blots were generated by stripping and reprobing of the same filter. B, mouse VEGF mRNA ELISA analysis of skin, liver, lung, bone marrow, spleen, lung, muscle, kidney, and heart tissues of SCID mice. The results are expressed as a ratio of the amount VEGF mRNA in DC101-treated mice to that in untreated mice. Lung tissues showed a higher expression of VEGF mRNA compared with other organs but no difference between DC101-treated and untreated mice; in contrast, skin, bone marrow, spleen, and heart tissues showed an increase in VEGF mRNA after DC101 treatment of mice.
Volume 64, Issue 18
