Article Figures & Data
Figures
- FIGURE 1.
Autoradiography and biodistribution studies on Granta-519 and SUDHL-4 tumors. (A) Autoradiography of tumor sections from immunocompromised mice was performed 24 h after injection of 125I-Fab-ABD or 125I-Fab-PAS200 or 1 h after injection of 18F-FDG. (B and C) Biodistribution studies of Fab-ABD and Fab-PAS200 labeled with 125I were conducted 24 h after injection. Tumor-to-organ ratios are plotted for both αCD20 Fabs (mean ± SD; n = 5). (D) No strong correlation between tumor uptake and tumor weight is found for SUDHL-4 tumors 24 h after injection, irrespective of the Fab radiotracer used (Pearson coefficient of −0.32 for Fab-PAS200 and −0.36 for Fab-ABD).
- FIGURE 2.
Histologic and immunohistochemical characterization of Granta-519 and SUDHL-4 tumors. (A) Explanted tumors were stained with hematoxylin and eosin (H&E) or immunostained for the target antigen CD20, apoptosis marker caspase 3 (Casp3), proliferation marker Mib1, or endothelial vessel marker CD31 (scale bar, 50 μm). (B) Example is shown of H&E-stained SUDHL-4 tumor comprising an area of 6 × 10 mm. (C) Quantification of Mib1 staining reveals significantly stronger proliferation for Granta-519 tumors than for SUDHL-4 tumors (P = 0.01). (D) Quantification of vessel area (n = 5) reveals significantly higher values for Granta-519 tumors than for SUDHL-4 tumors (P < 0.01).
- FIGURE 3.
Visualization of vessels and αCD20 Cy5-Fab-PAS200 distribution in Granta-519 and SUDHL-4 tumors. Both xenograft models were stained in vivo with fluorescence-labeled αCD20 Fab-PAS200 (24 h before sacrifice) and fluorescence-labeled B. (G.) simplicifolia-isolectin 1 for vessels (5 min before sacrifice) and imaged by light-sheet fluorescence microscopy. (A) Both xenograft models show extensive vascularization (red) throughout. Heterogeneous penetration and distribution of labeled Fab-PAS200 (green) is observed across entire Granta-519 tumor, whereas for the SUDHL-4 tumor nearly no Fab-PAS200 outside vessels is detected. Heat map visualization confirms this different distribution pattern of Granta-519 and SUDHL-4 tumors (scale bar, 1,500 μm). (B) Different localization of labeled Fab-PAS200 (green) in relation to lectin-labeled vessels (red) is clearly visible in magnified images of the maximum-intensity projection of 25 virtual single optical slices. Granta-519 tumor shows perivascular distribution of Fab-PAS200, whereas SUDHL-4 tumor shows confinement of Fab-PAS200 to intravascular space (arrows).
- FIGURE 4.
DCE MRI perfusion study of NHL xenograft models. (A) T2-weighted anatomic images are shown of an example Granta-519 tumor and an example SUDHL-4 tumor and their corresponding gadolinium-DTPA T1-weighted maps with color scale. (B) Gadolinium-concentration curves show higher wash-in pattern in individual Granta-519 tumors than in SUDHL-4 tumors despite considerable interindividual variation.
- FIGURE 5.
H33342 perfusion and penetration in Granta-519 and SUDHL-4 tumors. (A) Perfusion dye H33342 was injected 24 h after radioiodinated Fab-ABD or Fab-PAS200 and 1 min before mice were sacrificed. Afterward, CD20 (green) and CD31 (red) were detected in tumor cryosections using immunofluorescence. H33342 perfusion of 3 different SUDHL-4 and Granta-519 tumors is represented in blue. (B) Autoradiography and corresponding H33342 intensity is shown for an example SUDHL-4 tumor. (C) Mean H33342 penetration into tissue was quantified using distance maps based on CD31-positive vessels, with distance of zero equivalent to vessel lumen. H33342 intensity was normalized with regard to first distance value (25 μm) after vessel. 2F2 = CD20-specific Fab, derived from ofatumumab.
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