title: Monitoring of in vivo function of superparamagnetic iron oxide labelled murine dendritic cells during anti-tumour vaccination.
abstract: Dendritic cells (DCs) generated in vitro to present tumour antigens have been injected in cancer patients to boost in vivo anti-tumour immune responses. This approach to cancer immunotherapy has had limited success. For anti-tumour therapy, delivery and subsequent migration of DCs to lymph nodes leading to effective stimulation of effector T cells is thought to be essential. The ability to non-invasively monitor the fate of adoptively transferred DCs in vivo using magnetic resonance imaging (MRI) is an important clinical tool to correlate their in vivo behavior with response to treatment. Previous reports of superparamagnetic iron oxides (SPIOs) labelling of different cell types, including DCs, have indicated varying detrimental effects on cell viability, migration, differentiation and immune function. Here we describe an optimised labelling procedure using a short incubation time and low concentration of clinically used SPIO Endorem to successfully track murine DC migration in vivo using MRI in a mouse tumour model. First, intracellular labelling of bone marrow derived DCs was monitored in vitro using electron microscopy and MRI relaxometry. Second, the in vitro characterisation of SPIO labelled DCs demonstrated that viability, phenotype and functions were comparable to unlabelled DCs. Third, ex vivo SPIO labelled DCs, when injected subcutaneously, allowed for the longitudinal monitoring by MR imaging of their migration in vivo. Fourth, the SPIO DCs induced the proliferation of adoptively transferred CD4(+) T cells but, most importantly, they primed cytotoxic CD8(+) T cell responses to protect against a B16-Ova tumour challenge. Finally, using anatomical information from the MR images, the immigration of DCs was confirmed by the increase in lymph node size post-DC injection. These results demonstrate that the SPIO labelling protocol developed in this study is not detrimental for DC function in vitro and in vivo has potential clinical application in monitoring therapeutic DCs in patients with cancer.
authors.affiliation: Division of Imaging Sciences and Biomedical Engineering, Department of Imaging Chemistry and Biology, King's College London, St. Thomas' Hospital, London, United Kingdom.
authors.forename: Richard
authors.initials: R
authors.lastname: Tavaré
authors.forename: Pervinder
authors.initials: P
authors.lastname: Sagoo
authors.forename: Gopal
authors.initials: G
authors.lastname: Varama
authors.forename: Yakup
authors.initials: Y
authors.lastname: Tanriver
authors.forename: Alice
authors.initials: A
authors.lastname: Warely
authors.forename: Sandra S
authors.initials: SS
authors.lastname: Diebold
authors.forename: Richard
authors.initials: R
authors.lastname: Southworth
authors.forename: Tobias
authors.initials: T
authors.lastname: Schaeffter
authors.forename: Robert I
authors.initials: RI
authors.lastname: Lechler
authors.forename: Reza
authors.initials: R
authors.lastname: Razavi
authors.forename: Giovanna
authors.initials: G
authors.lastname: Lombardi
authors.forename: Gregory E D
authors.initials: GE
authors.lastname: Mullen
mesh_terms: D000818:Animals; D001681:Biological Assay; D001854:Bone Marrow Cells / Q000166:cytology; D019496:Cancer Vaccines / Q000276:immunology*; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D003287:Contrast Media / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000648:ultrastructure; D003911:Dextrans / Q000378:metabolism*; D008198:Lymph Nodes / Q000378:metabolism; D008279:Magnetic Resonance Imaging; D058185:Magnetite Nanoparticles; D008546:Melanoma, Experimental / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D010641:Phenotype; D013194:Staining and Labeling; D013601:T-Lymphocytes / Q000166:cytology; D014611:Vaccination
chemical_list: D019496:Cancer Vaccines; D003287:Contrast Media; D003911:Dextrans; D058185:Magnetite Nanoparticles; C065507:ferumoxides