Article Figures & Data
Figures
- FIGURE 1.
Assessment of reporter gene expression in vitro. (A) Subcellular localization of native-dCK/GFP, dCKDM/GFP, dCKTM/GFP, and ∆dCK/GFP reporter proteins. (B) Western blot analysis of dCK and mutant protein expression in nontransduced and transduced cell populations using human dCK and GFP- and β-actin–specific monoclonal antibodies. (C) 3H-FAC uptake (Ki) in nontransduced and transduced U87 cells is listed on abscissa. Values are mean ± SD, n = 3. N/T = nontransduced.
- FIGURE 2.
Radiotracer accumulation in transduced cells in vitro. Radiotracer uptake (Ki) of 3H-FEAU, 14C-FIAU, and 3H-penciclovir in nontransduced and transduced U87 cells is listed on abscissa. Values are mean ± SD, n = 3, P < 0.05. N/D = not done; N/T = nontransduced; PCV = penciclovir.
- FIGURE 3.
Small-animal PET of reporter gene expression. (A) Coronal small-animal PET images through xenografts placed subcutaneously over shoulders are shown: nontransduced and transduced U87 xenografts, including dCK, dCKDM, HSV1-tk, and HSV1-R176Qsr39tk. 18F-FEAU and 18F-FHBG images at 2 h after radiotracer administration obtained on consecutive days are shown for same animal. All images were adjusted to same color scale. (B) Image-based measurements of 18F-FEAU and 18F-FHBG at 2 h after radiotracer administration in same animals, expressed as percentage injected dose per cubic centimeter of tissue (%ID/cm3). Values are mean ± SD, n = 5. N/T = nontransduced.
- FIGURE 4.
Small-animal PET/micro-CT of dCKDM/GFP-expressing T cells: colocalization of 18F-FEAU accumulation in small-animal PET images with tumor regions in micro-CT images. Representative tumor-bearing animals from Pz1+dCKDM/GFP+ treated (A) and control nontreated (B) groups are shown. High 18F-FEAU accumulation was detected on day of T-cell administration in region corresponding to tumor foci on micro-CT image in Pz1+dCKDM/GFP+ treated animal. No 18F-FEAU accumulation was observed in region corresponding to tumor foci in nontreated control animal. Heart and blood vessels were enhanced by iodinated lipids. Immunofluorescent analysis revealed presence of GFP-positive (green, stained with anti-GFP antibody) T cells infiltrating hPSMA-positive (red, stained with anti-hPSMA antibody) tumor in lung from representative Pz1+dCKDM/GFP+ T-cell–treated (C) and –nontreated tumor-bearing (D) animals. Nuclei were stained with 4,6-diamidino-2-phenylindole (blue). Magnification, ×10. Quantitative analysis of 18F-FEAU-accumulation (E) and region-of-interest–to–background ratios (F) in treated and nontreated groups is presented. Data are expressed as percentage injected dose per cubic centimeter of tissue (%ID/cm3). Values are mean ± SD, n = 5. BGR = background; ROI = region of interest, marked with arrow.
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