DENDRITIC CELLS IN A MURINE MODEL OF BREAST CANCER: FUNCTIONAL AND PHENOTYPIC EVALUATION USING NANOPARTICLES
type: CreativeWork
description: Dendritic cells play a pivotal role in antigen presentation and adaptive immune response activation. Studies on mice models reported that tumoral antigens-loaded DCs can induce therapeutic and protective anti-tumour immune responses, both in vitro and in vivo. These results provide an evidence of the possible use of DCs as vaccines. However, the efficacy of such therapeutic vaccination was recently questioned due to the limited degree of actual tumour regression in some clinical trials. Thus, it is necessary to better characterize immunological and clinical parameters of DCs immunization to increase the therapeutic efficay of the vaccines. In vivo imaging tecniques, as magnetic resonance imaging (MRI) and single photon emission tomography (SPET), are used to study cell trafficking mechanisms in vivo and state anti-tumoral treatments efficacy, allowing an accurate assessment. In this work we initially set the optimal culture conditions to obtain functional immature DCs. Evaluations based on DCs number, cell viability and immature DCs percentage led to establish day 6 as the one in which the highest number of functionally active immature DCs can be achieved. We then set a DCs labelling protocol with commercial SPIO (Endorem®) evaluating Iron content using Perl’s staining and relassometry. Time and dose-response kinetic evaluations allowed to define as optimal labelling conditions the use of 200 μg Iron of Endorem®/ ml of culture for 16 hours. Cell viability and phenotypic markers analysis proved that SPIO labelling had no influence on immature DCs viability or activation status. Images acquired by transmission electron microscopy showed that Endorem® were inside cells, in particular inside lysosomes, and were taken up by endocytosis mechanisms. We then focused on DCs antigens loading. Tumoral antigens were obtained lysing neoplastic masses extracted from a mammary tumour mouse model (MMTV-Ha-Ras). DCs pulsed with different antigen concentrations resulted mature by mea…
abstract: Dendritic cells play a pivotal role in antigen presentation and adaptive immune response activation. Studies on mice models reported that tumoral antigens-loaded DCs can induce therapeutic and protective anti-tumour immune responses, both in vitro and in vivo. These results provide an evidence of the possible use of DCs as vaccines. However, the efficacy of such therapeutic vaccination was recently questioned due to the limited degree of actual tumour regression in some clinical trials. Thus, it is necessary to better characterize immunological and clinical parameters of DCs immunization to increase the therapeutic efficay of the vaccines. In vivo imaging tecniques, as magnetic resonance imaging (MRI) and single photon emission tomography (SPET), are used to study cell trafficking mechanisms in vivo and state anti-tumoral treatments efficacy, allowing an accurate assessment. In this work we initially set the optimal culture conditions to obtain functional immature DCs. Evaluations based on DCs number, cell viability and immature DCs percentage led to establish day 6 as the one in which the highest number of functionally active immature DCs can be achieved. We then set a DCs labelling protocol with commercial SPIO (Endorem®) evaluating Iron content using Perl’s staining and relassometry. Time and dose-response kinetic evaluations allowed to define as optimal labelling conditions the use of 200 μg Iron of Endorem®/ ml of culture for 16 hours. Cell viability and phenotypic markers analysis proved that SPIO labelling had no influence on immature DCs viability or activation status. Images acquired by transmission electron microscopy showed that Endorem® were inside cells, in particular inside lysosomes, and were taken up by endocytosis mechanisms. We then focused on DCs antigens loading. Tumoral antigens were obtained lysing neoplastic masses extracted from a mammary tumour mouse model (MMTV-Ha-Ras). DCs pulsed with different antigen concentrations resulted mature by mea…
author: M. Borelli
language: eng
keywords: Dendritic Cell, Breast cancer, Nanoparticles, Settore MED/04 - Patologia Generale
date published: 2010-12-09
date modified: 2026-07-27T20:19:17Z
provider: Università degli Studi di Milano
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: Università degli Studi di Milano

PropertyValue
property ID: Handle
value: http://hdl.handle.net/2434/150209

PropertyValue
property ID: DOI
value: 10.13130/borelli-manuela_phd2010-12-09