In Vivo Imaging of Lymph Node Migration of MNP- and 111In-Labeled Dendritic Cells in a Transgenic Mouse Model of Breast Cancer (MMTV-Ras)

PurposeThe authors present a protocol for the in vivo evaluation, using different imaging techniques, of lymph node (LN) homing of tumor-specific dendritic cells (DCs) in a murine breast cancer model.ProceduresBone marrow DCs were labeled with paramagnetic nanoparticles (MNPs) or 111In-oxine. Antigen loading was performed using tumor lysate. Mature DCs were injected into the footpads of transgenic tumor-bearing mice (MMTV-Ras) and DC migration was tracked by magnetic resonance imaging (MRI) and single-photon emission computed tomography (SPECT). Ex vivo analyses were performed to validate the imaging data.ResultsDC labeling, both with MNPs and with 111In-oxine, did not affect DC phenotype or functionality. MRI and SPECT allowed the detection of iron and 111In in both axillary and popliteal LNs. Immunohistochemistry and γ-counting revealed the presence of DCs in LNs.ConclusionsMRI and SPECT imaging, by allowing in vivo dynamic monitoring of DC migration, could further the development and optimization of efficient anti-cancer vaccines.

[1]  E T Ahrens,et al.  Receptor‐mediated endocytosis of iron‐oxide particles provides efficient labeling of dendritic cells for in vivo MR imaging , 2003, Magnetic resonance in medicine.

[2]  T. Scheenen,et al.  Sensitivity of magnetic resonance imaging of dendritic cells for in vivo tracking of cellular cancer vaccines , 2006, International journal of cancer.

[3]  A. Devillers,et al.  Biodistribution of radiolabelled human dendritic cells injected by various routes , 2005, European Journal of Nuclear Medicine and Molecular Imaging.

[4]  G. Lucignani,et al.  Molecular imaging of cell-mediated cancer immunotherapy. , 2006, Trends in biotechnology.

[5]  Hisataka Kobayashi,et al.  In vivo real-time, multicolor, quantum dot lymphatic imaging. , 2009, The Journal of investigative dermatology.

[6]  H. Soltau,et al.  The DRAGO gamma camera. , 2010, The Review of scientific instruments.

[7]  G. Schuler,et al.  An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. , 1999, Journal of immunological methods.

[8]  P. Foster,et al.  In Vivo Cellular MRI of Dendritic Cell Migration Using Micrometer-Sized Iron Oxide (MPIO) Particles , 2011, Molecular Imaging and Biology.

[9]  F. Sallusto,et al.  The instructive role of dendritic cells on T cell responses: lineages, plasticity and kinetics. , 2001, Current opinion in immunology.

[10]  A. Moretti,et al.  Evaluation of in vivo labelled dendritic cell migration in cancer patients , 2004, Journal of Translational Medicine.

[11]  Masafumi Nakamura,et al.  Combination therapy with tumor cell-pulsed dendritic cells and activated lymphocytes for patients with disseminated carcinomas. , 2005, Anticancer research.

[12]  S. Aamdal,et al.  Phase I/II trial of melanoma therapy with dendritic cells transfected with autologous tumor-mRNA , 2006, Cancer Gene Therapy.

[13]  Yi Lu,et al.  Smart “Turn‐on” Magnetic Resonance Contrast Agents Based on Aptamer‐Functionalized Superparamagnetic Iron Oxide Nanoparticles , 2007, Chembiochem : a European journal of chemical biology.

[14]  P. Leder,et al.  Animal models of human disease. Pathology and molecular biology of spontaneous neoplasms occurring in transgenic mice carrying and expressing activated cellular oncogenes. , 1989, The American journal of pathology.

[15]  E. Ahrens,et al.  Functional assessment of human dendritic cells labeled for in vivo (19)F magnetic resonance imaging cell tracking. , 2010, Cytotherapy.

[16]  J. Timmerman,et al.  Dendritic cell vaccines for cancer immunotherapy. , 1999, Annual review of medicine.

[17]  Ralph Weissleder,et al.  Near-infrared fluorescent nanoparticles as combined MR/optical imaging probes. , 2002, Bioconjugate chemistry.

[18]  H. Johnsen,et al.  Vaccination with p53 peptide-pulsed dendritic cells is associated with disease stabilization in patients with p53 expressing advanced breast cancer; monitoring of serum YKL-40 and IL-6 as response biomarkers , 2007, Cancer Immunology, Immunotherapy.

[19]  R. Levy,et al.  Production of myeloid dendritic cells (DC) pulsed with tumor-specific idiotype protein for vaccination of patients with multiple myeloma. , 2006, Cytotherapy.

[20]  Xiaoyuan Chen,et al.  Nanoparticles for cell labeling. , 2011, Nanoscale.

[21]  Kazushi Inoue,et al.  MMTV mouse models and the diagnostic values of MMTV-like sequences in human breast cancer , 2009, Expert review of molecular diagnostics.

[22]  Keith L. Black,et al.  Vaccination with Tumor Lysate-Pulsed Dendritic Cells Elicits Antigen-Specific, Cytotoxic T-Cells in Patients with Malignant Glioma , 2004, Cancer Research.

[23]  J. Banchereau,et al.  Longitudinal tracking of human dendritic cells in murine models using magnetic resonance imaging , 2010, Magnetic resonance in medicine.

[24]  S. Laurent,et al.  Magnetic labeling of non-phagocytic adherent cells with iron oxide nanoparticles: a comprehensive study. , 2008, Contrast media & molecular imaging.

[25]  H. Lyerly,et al.  Dendritic cell vaccines. , 2007, Frontiers in bioscience : a journal and virtual library.

[26]  R. Weissleder,et al.  Cell Internalization of Magnetic Nanoparticles Using Transfection Agents , 2007, Molecular imaging.

[27]  M. Erturk,et al.  Active immunotherapy for cancer patients using tumor lysate pulsed dendritic cell vaccine: a safety study. , 2007, Journal of experimental & clinical cancer research : CR.

[28]  H. Ueno,et al.  Dendritic Cells Loaded With Killed Allogeneic Melanoma Cells can Induce Objective Clinical Responses and MART-1 Specific CD8+ T-cell Immunity , 2006, Journal of immunotherapy.

[29]  W. Van den Broeck,et al.  Anatomy and nomenclature of murine lymph nodes: Descriptive study and nomenclatory standardization in BALB/cAnNCrl mice. , 2006, Journal of immunological methods.

[30]  Ralph Weissleder,et al.  Tat peptide-derivatized magnetic nanoparticles allow in vivo tracking and recovery of progenitor cells , 2000, Nature Biotechnology.

[31]  Z. Gu,et al.  Self-assembly of magnetite nanocrystals with amphiphilic polyethylenimine: structures and applications in magnetic resonance imaging. , 2009, Journal of nanoscience and nanotechnology.

[32]  Antonio Lanzavecchia,et al.  Regulation of T Cell Immunity by Dendritic Cells , 2001, Cell.

[33]  J. Gore,et al.  Magnetic nanoparticles for imaging dendritic cells , 2010, Magnetic resonance in medicine.

[34]  C. Corot,et al.  Superparamagnetic agents: physicochemical characteristics and preclinical imaging evaluation. , 1996, Academic radiology.

[35]  Jinming Gao,et al.  Nonclustered magnetite nanoparticle encapsulated biodegradable polymeric micelles with enhanced properties for in vivo tumor imaging , 2011 .

[36]  E. Mittendorf,et al.  Evaluation of the HER2/neu‐derived peptide GP2 for use in a peptide‐based breast cancer vaccine trial , 2006, Cancer.

[37]  B. Neyns,et al.  Current approaches in dendritic cell generation and future implications for cancer immunotherapy , 2007, Cancer Immunology, Immunotherapy.

[38]  Andreas Hess,et al.  In vivo magnetic resonance imaging of dendritic cell migration into the draining lymph nodes of mice , 2006, European journal of immunology.

[39]  Parag Aggarwal,et al.  Preclinical studies to understand nanoparticle interaction with the immune system and its potential effects on nanoparticle biodistribution. , 2008, Molecular pharmaceutics.

[40]  M. Lutz,et al.  Non-invasive imaging of dendritic cell migration in vivo. , 2006, Immunobiology.

[41]  J. Fay,et al.  Dendritic cell based tumor vaccines. , 2000, Immunology letters.

[42]  R. Maldonado-López,et al.  Dendritic cell subsets and the regulation of Th1/Th2 responses. , 2001, Seminars in immunology.

[43]  Arend Heerschap,et al.  Magnetic resonance tracking of dendritic cells in melanoma patients for monitoring of cellular therapy , 2005, Nature Biotechnology.

[44]  W. Ge,et al.  Heat-shocked tumor cell lysate-pulsed dendritic cells induce effective anti-tumor immune response in vivo. , 2006, World journal of gastroenterology.

[45]  E. Zappi,et al.  Combined Fontana-Masson/Perls' staining. , 1984, The American Journal of dermatopathology.

[46]  G. Hager,et al.  Glucocorticoid regulation of the Ha-MuSV p21 gene conferred by sequences from mouse mammary tumor virus , 1981, Cell.

[47]  Donald S. Williams,et al.  Detection of single mammalian cells by high-resolution magnetic resonance imaging. , 1999, Biophysical journal.

[48]  J. Gore,et al.  Dendritic cells: therapy and imaging , 2009, Expert opinion on biological therapy.

[49]  E. Gilboa DC-based cancer vaccines. , 2007, The Journal of clinical investigation.

[50]  V. Cerundolo,et al.  Dendritic cells: a journey from laboratory to clinic , 2004, Nature Immunology.

[51]  J. Hay,et al.  Measurement of lymphocyte traffic with indium-111. , 1980, Clinical and experimental immunology.

[52]  Stephanie E. A. Gratton,et al.  The effect of particle design on cellular internalization pathways , 2008, Proceedings of the National Academy of Sciences.

[53]  T. C. Jones,et al.  ANIMAL MODELS OF HUMAN DISEASE , 2009 .

[54]  S. Hudak,et al.  The CC Chemokine Receptor-7 Ligands 6Ckine and Macrophage Inflammatory Protein-3β Are Potent Chemoattractants for In Vitro- and In Vivo-Derived Dendritic Cells , 1999, The Journal of Immunology.

[55]  J. Villadangos,et al.  Migratory dendritic cells transfer antigen to a lymph node-resident dendritic cell population for efficient CTL priming. , 2006, Immunity.