Comparison of three different methods for radiolabelling human activated T lymphocytes

Carlo Botti, Donatella R M Negri, Ettore Seregni, Venkatesh Ramakrishna, Flavio Arienti, Lorenzo Maffioli, Claudia Lombardo, Anna Bogni, Claudio Pascali, Flavio Crippa, Simonetta Massaron, Federica Remonti, Silvia Nerini-Molteni, Silvana Canevari, Emilio Bombardieri

Research output: Contribution to journalArticlepeer-review


One approach in the treatment of ovarian cancer patients involves the infusion of autologous T lymphocytes coupled with a bispecific monoclonal antibody MOv18/anti-CD3 (biMAb OC/TR), which recognizes a 38-kDa glycoprotein expressed on ovarian carcinomas and the CD3 T cell receptor. However, little is known about the in vivo biodistribution of injected activated lymphocytes, information that could be obtained by scintigraphic imaging of radiolabelled T cells in order to visualize the migratory pattern. We compared the efficiency, stability and toxicity of technetium-99m hexamethylpropylene amine oxime (HMPAO), indium-111 oxine and fluorine-18 2-fluoro-2-deoxy-D-glucose (FDG) in radiolabelling activated lymphocytes targeted with biMAb OC/TR. The mean labelling efficiencies of 111In-oxine and 18F-FDG using 2.5 x 108 lymphocytes (68% and 64%, respectively) were more than twice that of 99mTc-HMPAO (31%). Retention of the radionuclide in the cell was highest in the case of 111In-oxine labelling (less than 25% of the initial cell-bound activity released after 240 min, as compared with 44% of the 99mTc label in the same period and 45% of 18F radionuclide released after 150 min). None of the three radiolabelling reagents induced any significant alteration in cell viability or immunophenotype. However, both 111In-oxine and 18F-FDG induced a loss of cytotoxic activity of lymphocytes against the ovarian carcinoma cell line IG-ROV1, and all three radiolabelling reagents caused a significant reduction in the proliferative ability of labelled lymphocytes compared to controls, with cell death occurring after 8-9 days. Radiolabelling with the more stable 111In-oxine reagent using a higher number of lymphocytes (1.4 x 109) but the same total activity (around 55.5 MBq) resulted in improved labelled T cell viability and proliferative ability, although the mean labelling efficiency decreased (35.8%). Together the data suggest that 111In-oxine at low activity per cell is the most appropriate reagent for radiolabelling activated retargeted T lymphocytes useful for in vivo biodistribution studies.

Original languageEnglish
Pages (from-to)497-504
Number of pages8
JournalEuropean Journal Of Nuclear Medicine
Issue number5
Publication statusPublished - 1997


  • Adoptive immunotherapy
  • Bispecific monoclonal antibody
  • Fluorine-18 2-fluoro-2-deoxy-D-glucose
  • Indium-111 oxine
  • Lymphocyte radiolabelling
  • Ovarian cancer
  • Technetium hexamethylpropylene amine oxime

ASJC Scopus subject areas

  • Radiology Nuclear Medicine and imaging


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