Research ArticleBasic Science Investigation

Trafficking Macrophage Migration Using Reporter Gene Imaging with Human Sodium Iodide Symporter in Animal Models of Inflammation

Ji Hyoung Seo, Yong Hyun Jeon, Yong Jin Lee, Gil Sook Yoon, Dong-Il Won, Jeoung-Hee Ha, Shin Young Jeong, Sang Woo Lee, Byeong Cheol Ahn and Jaetae Lee
Journal of Nuclear Medicine October 2010, 51 (10) 1637-1643; DOI: https://doi.org/10.2967/jnumed.110.077891

Article Figures & Data

Figures

  • FIGURE 1.
    FIGURE 1.

    Establishment of macrophage cell line coexpressing hNIS and GFP genes. Immunofluorescent staining of RNIS cells was performed using anti-hNIS antibody and Alexa Fluor 568–conjugated secondary antibody to determine hNIS protein expression. Detailed experimental methods are described in “Materials and Methods” section. Ab = antibody; DAPI = 4′-6-diamidino-2-phenylindole.

  • FIGURE 2.
    FIGURE 2.

    Validation of hNIS expression in vitro. (A) In vitro kinetics of 125I in macrophage cells expressing hNIS gene are presented. Differences in 125I uptake are according to cell numbers and 125I efflux study in RNIS cells. Half-time (T1/2) of 125I efflux was about 5 min. Detailed experimental methods are described in “Materials and Methods” section. Data are expressed as mean ± SD. (B) Scintigraphic imaging of nude mice implanted with RNIS cells. RNIS (8 × 106 and 2 × 107) cells were subcutaneously inoculated in right and left thighs of nude mice, respectively. Twenty-four hours later, scintigraphic imaging was acquired after intravenous injection of 18.5 MBq of 99mTc-pertechnetate in nude mice. In both thighs of mice inoculated with RNIS cells, there was focal tracer accumulation proportional to increases in cell numbers. Physiologic iodide uptake was shown in thyroid, stomach, and urinary bladder. B = bladder; CPM = counts per minute; S = stomach; T = thyroid.

  • FIGURE 3.
    FIGURE 3.

    Effect of reporter gene transduction on proliferation and cytokine production (by lipopolysaccharide treatment) of macrophage cells. Cells were stimulated with lipopolysaccharide (1 μg/mL), and then 24 h later, culture supernatant was analyzed to determine amount of IL-12p70 and TNF-α using enzyme-linked immunosorbent assay. Data are expressed as mean ± SD. LPS = lipopolysaccharide.

  • FIGURE 4.
    FIGURE 4.

    Small-animal PET in focal inflammation model. (A) Representative photograph of inflammation model induced by turpentine oil. Yellow arrows indicate stripped inflammatory tissue. (B) Small-animal PET images of each mouse group administered parental RAW264.7 or RNIS cells. (Left) 18F-FDG PET scan demonstrates donut-shaped hot spot at inflammation sites of mice administered RAW264.7 and RNIS cells. (Right) 124I PET scan also clearly demonstrates donut-shaped focal uptake at inflammation site of mice administered RNIS cells. Hot spot of PET images with 124I correlated strongly with that of 18F-FDG PET images at inflammation site of mice administered RNIS cells, but tracer uptake was not observed at inflammation site of mice injected with parental RAW264.7 cells. (C) Iodine accumulation was 2.17 times higher in region of interest drawn over inflammation site of mice injected with RNIS cells. %ID/g = percentage injected dose per gram.

  • FIGURE 5.
    FIGURE 5.

    Immunohistochemical analysis of hNIS and GFP genes. In inflammation site of mice administered RAW264.7 cells, skeletal muscle showed extensive coagulation necrosis with viable foamy macrophages. By immunohistochemistry, RAW264.7 cells revealed negative reactivity for GFP and NIS (×400). In inflammation site of mice administered RNIS cells, cells showed diffuse and strong positive staining for GFP with cytoplasmic patterns. Also, macrophages revealed diffuse and strong positivity for NIS with cytoplasmic membrane pattern (×400).

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