Article Figures & Data
Figures
- FIGURE 1.
Flow cytometric analysis of EGFR expression on HNSCC cells. Panitumumab was used as primary antibody, and FITC-conjugated donkey antihuman IgG was used as secondary antibody. Mean values (±SD) of FITC signal intensity (MFI) of 3 measurements are shown. 22B = UM-SCC-22B.
- FIGURE 2.
(A) Small-animal PET images of HNSCC tumor-bearing nude mice at different time points after intravenous injection of 64Cu-DOTA-panitumumab (n = 4/group). Decay-corrected transaxial images at different time points are shown, and tumors are indicated by arrowheads. For UM-SCC-22B and SAS tumors, scale ranged from 0 %ID/g to 30 %ID/g, and for SQB20 tumors, scale ranged from 0 %ID to 15 %ID/g for optimal visualization. (B) HNSCC tumor uptake levels of 64Cu-DOTA-panitumumab and 64Cu-DOTA-IgG at 20 h after injection quantified from small-animal PET scans (n = 4). 22B = UM-SCC-22B. *P < 0.05.
- FIGURE 3.
Immunofluorescence examination of EGFR expression and panitumumab localization in HNSCC tumors. Images were obtained under same conditions and displayed at same magnification and scale (except D). (A) Tumor sections were directly stained with panitumumab as primary antibody and with FITC-conjugated donkey antihuman IgG as secondary antibody. Murine CD31 was stained with Cy3-conjugated IgG to visualize tumor vasculature. SQB20 tumors showed highest fluorescence intensity, corresponding to highest EGFR expression. (B) Thirty hours after FITC-panitumumab injection, tumors were harvested and tumor sections were observed after being mounted with DAPI-containing medium. (C) Thirty hours after DOTA-panitumumab injection, tumors were harvested and tumor sections were stained with FITC-conjugated donkey antihuman IgG. (D) In high-magnification view of images shown in C, color was rescaled to emphasize relationship of panitumumab and vasculature (red from Cy3 for CD31; green from FITC for EGFR and panitumumab; blue from DAPI for nucleus visualization).
- FIGURE 4.
MVD measurement of HNSCC tumors. Frozen slices of HNSCC tumor were stained with rat antimouse CD31 antibody and visualized using Cy3-conjugated rat antimouse IgG. After CD31 staining, 10 random views in both center and periphery of tumor slices were selected for MVD analysis using observer-set threshold to distinguish vascular elements from surrounding tissue parenchyma. Number of vessels counted was divided by field of view to yield MVD, as number of vessels/mm2. 22B = UM-SCC-22B. *P < 0.05. *P < 0.05. **P < 0.01.
- FIGURE 5.
Measurement of vascular permeability using Evans blue. Tumors were excised 4 h after dye had been injected. After being mounted with medium containing DAPI, sections were observed under LSM 510 (Zeiss) confocal microscope with optical filters (543-nm excitation; long pass, 585-nm emission). For quantification, Evans blue was extracted in formamide (0.01 mL/mg of tumor tissue) for 72 h. Relative Evans blue concentrations were determined by measuring light absorbance at 620 nm. 22B = UM-SCC-22B. *P < 0.05. **P < 0.01.
Tables
64Cu-DOTA-panitumumab (%ID/g) 64Cu-DOTA-IgG (%ID/g) Site 4 h 20 h 30 h 48 h 4 h 20 h 30 h 48 h Blood 22.73 ± 3.99 14.42 ± 4.47 12.35 ± 4.25 10.29 ± 4.04 20.54 ± 2.58 13.84 ± 2.24 13.01 ± 1.28 11.49 ± 1.99 Liver 15.94 ± 3.35 12.52 ± 3.29 11.96 ± 3.87 11.55 ± 3.64 13.77 ± 2.04 10.42 ± 1.17 10.92 ± 1.77 10.83 ± 2.25 Muscle 3.32 ± 1.50 2.479 ± 1.08 2.79 ± 1.31 2.14 ± 1.03 2.63 ± 0.78 2.20 ± 0.40 2.72 ± 0.83 2.10 ± 0.11 22B 16.09 ± 6.52 26.41 ± 9.16 31.42 ± 10.77 34.80 ± 9.26 5.70 ± 3.92 10.27 ± 5.27 11.75 ± 6.35 12.14 ± 6.71 SAS 6.39 ± 1.60* 11.01 ± 1.84* 12.39 ± 4.15* 15.35 ± 3.33* 3.51 ± 0.86 6.26 ± 0.68 7.20 ± 2.25 8.05 ± 3.75 SQB20 4.02 ± 1.87† 7.92 ± 1.48† 8.76 ± 1.07† 9.39 ± 1.44† 3.54 ± 1.04 5.07 ± 2.30 7.26 ± 5.15 8.57 ± 5.04
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