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Synthesis of 99mTc-HYNIC-interleukin-12, a new specific radiopharmaceutical for imaging T lymphocytes

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Abstract

Purpose

Few radiopharmaceuticals have been described for the study of lymphocyte trafficking despite its high clinical relevance. The main difficulty resides in the identification of a suitable highly specific probe to target these cells. Interleukin-12 (IL12) is a heterodimeric cytokine which plays a key role in the development of Th1 lymphocytes. The aims of the present study were to label IL12 with 99mTc, to evaluate its ability to bind to activated T lymphocytes in vitro and to study its biodistribution in normal mice and mice affected by autoimmune colitis.

Methods

IL12 was derivatised with HYNIC-NHS and labelled with 99mTc. An in vitro binding assay was performed on KIT225 cells, an IL12 receptor-positive cell line. 99mTc-IL12 biodistribution in normal mice was studied. Targeting experiments were performed in Balb/c mice injected with KIT225 cells and in mice with chemically induced chronic colitis.

Results

99mTc-IL12 labelling efficiency ranged between 75% and 85%. Saturation binding analysis revealed a K d of 2.09 nM. Results of biodistribution studies showed a predominant hepatic route of excretion. A significant degree of uptake in the spleen and thymus was also observed. In mice injected with KIT225 cells, 99mTc-IL12-specific uptake in these cells increased over time. 99mTc-IL12 also accumulated significantly in bowel of mice affected by TNBS-induced colitis showing T lymphocyte infiltration at histology, while accumulation in colon from control animals was negligible.

Conclusion

We conclude that this radiolabelled cytokine is a suitable candidate for specific in vivo imaging of T lymphocytes: a step forward in molecular imaging of immune-mediated processes.

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Acknowledgements

Dr. Alessio Annovazzi has been the recipient of a fellowship from the Institute Pasteur/Cenci-Bolognetti Foundation. This work was supported by the International Society of Radiolabelled Blood Elements (ISORBE), by the Nuklearmedizine Society e.V. and by the Italian Ministry of University and Research (MURST, “Progetto Giovani Ricercatori”). The authors wish to thank Wyeth Research (Cambridge, MA, USA) for the generous gift of recombinant hIL12. This work was performed in part within the COST B12 framework.

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Authors and Affiliations

  1. Nuclear Medicine, Ospedale S. Andrea, 2nd Faculty of Medicine “La Sapienza”, Via di Grottarossa 1035, 00189, Roma, Italy

    Alessio Annovazzi, Calogero D’Alessandria, Francesco Scopinaro & Alberto Signore

  2. Department of Biopathology and Diagnostic Imaging, University “Tor Vergata”, Rome, Italy

    Elena Bonanno

  3. Cancer Research UK, St. Bartholomew’s Hospital, London, UK

    Stephen J. Mather

  4. Division of Nuclear Medicine, University of Gent, Gent, Belgium

    Bart Cornelissen & Christophe van de Wiele

  5. Department of Nuclear Medicine, University of Groningen, Groningen, The Netherlands

    Alessio Annovazzi, Rudi A. Dierckx & Alberto Signore

  6. Department of Biology, NSTA, University “Tor Vergata”, Rome, Italy

    Maurizio Mattei & Graziana Palmieri

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  1. Alessio Annovazzi
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  2. Calogero D’Alessandria
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Correspondence to Alberto Signore.

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Annovazzi, A., D’Alessandria, C., Bonanno, E. et al. Synthesis of 99mTc-HYNIC-interleukin-12, a new specific radiopharmaceutical for imaging T lymphocytes. Eur J Nucl Med Mol Imaging 33, 474–482 (2006). https://doi.org/10.1007/s00259-005-0001-6

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  • DOI: https://doi.org/10.1007/s00259-005-0001-6

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