Serial in vivo imaging of the targeted migration of human HSV-TK-transduced antigen-specific lymphocytes

Abstract

New technologies are needed to characterize the migration, survival, and function of antigen-specific T cells in vivo. Here, we demonstrate that Epstein-Barr virus (EBV)–specific T cells transduced with vectors encoding herpes simplex virus-1 thymidine kinase (HSV-TK) selectively accumulate radiolabeled 2′-fluoro-2′-deoxy-1-β-D-arabinofuranosyl-5-iodouracil (FIAU). After adoptive transfer, HSV-TK+ T cells labeled in vitro or in vivo with [131I]FIAU or [124I]FIAU can be noninvasively tracked in SCID mice bearing human tumor xenografts by serial images obtained by scintigraphy or positron emission tomography (PET), respectively. These T cells selectively accumulate in EBV+ tumors expressing the T cells' restricting HLA allele but not in EBV or HLA-mismatched tumors. The concentrations of transduced T cells detected in tumors and tissues are closely correlated with the concentrations of label retained at each site. Radiolabeled transduced T cells retain their capacity to eliminate targeted tumors selectively. This technique for imaging the migration of ex vivo–transduced antigen-specific T cells in vivo is informative, nontoxic, and potentially applicable to humans.

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Figure 1: In vitro analyses of HSV-TK-transduced T lymphocytes.
Figure 2: HLA-restricted cytotoxicity of EBV-specific HSV-TK-transduced T cells and anti-EBV cytotoxic activity of T cells transduced to express HSV-TK after co-incubation with [124I]FIAU or [131I]FIAU.
Figure 3: Biodistribution of [131I]FIAU-labeled NIT+ human EBV-specific T cells over time in SCID mice bearing human EBV lymphoma xenografts.
Figure 4: MicroPET imaging of T-cell migration and targeting.
Figure 5: Tissue accumulation of [124I]FIAU by CTL-TKGFP and normal organs over the course of the experiment.

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Acknowledgements

This work was supported by US National Institutes of Health grants CA59350, P50 CA86438, CA23766, HL53752, CA57599, CA76117, R24 CA83084, Department of Energy grants FG02-02ER63481, FG03-86ER60407, and 95ER62039, a Translational Research Award of the Leukemia and Lymphoma Society, The Aubrey Fund for Pediatric Cancer Research, The Larry H. Smead Fund, and The Vincent Astor Chair Research Fund. We thank Judith Guerrero, Brad Beatty, and Tatiana Beresten for their excellent technical support.

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Correspondence to Richard J. O'Reilly.

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Koehne, G., Doubrovin, M., Doubrovina, E. et al. Serial in vivo imaging of the targeted migration of human HSV-TK-transduced antigen-specific lymphocytes. Nat Biotechnol 21, 405–413 (2003). https://doi.org/10.1038/nbt805

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