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Autor/a:
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Crisci, Elisa; Fraile Sauce, Lorenzo José; Novellas, Rosa; Espada, Yvonne; Cabezón, Raquel; Martínez, Jorge; Cordoba, Lorena; Bárcena, Juan; Benitez-Ribas, Daniel; Montoya, Maria
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Notas:
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Cellular therapies using immune cells and in particular dendritic cells (DCs) are being increasingly applied in clinical trials and vaccines. Their success partially depends on accurate delivery of cells to target organs or migration to lymph nodes. Delivery and subsequent migration of cells to regional lymph nodes is essential for effective stimulation of the immune system. Thus, the design of an optimal DC therapy would be improved by optimizing technologies for monitoring DC trafficking. Magnetic resonance imaging (MRI) represents a powerful tool for non-invasive imaging of DC migration in vivo. Domestic pigs share similarities with humans and represent an excellent animal model for immunological studies. The aim of this study was to investigate the possibility using pigs as models for DC tracking in vivo. Porcine monocyte derived DC (MoDC) culture with superparamagnetic iron oxide (SPIO) particles was standardized on the basis of SPIO concentration and culture viability. Phenotype, cytokine production and mixed lymphocyte reaction assay confirmed that porcine SPIO-MoDC culture were similar to mock MoDCs and fully functional in vivo. Alike, similar patterns were obtained in human MoDCs. After subcutaneous inoculation in pigs, porcine SPIO-MoDC migration to regional lymph nodes was detected by MRI and confirmed by Perls staining of draining lymph nodes. Moreover, after one dose of virus-like particles-pulsed MoDCs specific local and systemic responses were confirmed using ELISPOT IFN-γ in pigs. In summary, the results in this work showed that after one single subcutaneous dose of pulsed MoDCs, pigs were able to elicit specific local and systemic immune responses. Additionally, the dynamic imaging of MRI-based DC tracking was shown using SPIO particles. This proof-of-principle study shows the potential of using pigs as a suitable animal model to test DC trafficking with the aim of improving cellular therapies.
We want to thank: Ferrán López, Rosa López, Zoraida Cervera, Pamela Martinez-Orellana, Tufaria Mussá, Massimiliano Baratelli, Diego Pérez, Sergio López from CRESA and José Luis Ruiz de la Torre and Javier Aceña (UAB) for farm and technical support; Jaume Martorell (Fundació Hospital Clínic Veterinari, UAB) for MRI support; Javier Domínguez (INIA) for the porcine antibodies; Antonio Lestuzzi, Michele Crisci and Raif Yucel for MR imaging support; Joaquim Segalés for anatomic pathology analysis; Mónica Pérez for immunohistochemical stainings; Aida Neira and Blanca Pérez for Perls staining; Eva Huerta y Marina Sibila for PCV2 PCR; David Andreu and Beatriz García de la Torre (Pompeu Fabra University, Barcelona), and Esther Blanco (CISA-INIA, Madrid), for the FMDV 3A peptide; Alicia Solórzano for critically reviewing the manuscript. This work was funded by the project AGL2010-22200-C02 of Spanish Ministry of Science and Innovation. PhD studies of Raquel Cabezón are funded by a doctoral FI fellowship from the Generalitat de Catalunya. |
