Dendritic cell (DC)-based vaccines have shown promising therapeutic results in cancer and some immune disorders. It is critical to track in vivo migration behaviours of DCs and monitor the whole process dynamically and non-invasively. Superparamagnetic iron oxide (SPIO) nanoparticles are chosen for DC labelling under magnetic resonance imaging (MRI) because of their proven biosafety as contrast agents. However, when used for cell labelling, sensitive biological indicators such as cell autophagy may be helpful to better understand the process and improve the probe design. Here, lactosylated N-Alkyl polyethylenimine coated SPIO nanoparticles are used for DC labelling. This probe shows satisfactory cell labelling efficiency and low cytotoxicity. In this study, autophagy was used as a key factor to understand how DCs react to nanoparticles after labelling. Our results demonstrate that the nanoparticles can induce protective autophagy in DCs, as inhibition of the autophagy flux could lead to cell death. Meanwhile, the nanoparticles induced autophagy could promote DC maturation which is an essential process for its migration and antigen presentation. Autophagy induced DC maturation is known to enhance the vaccine functions of DCs, therefore, our results suggest that beyond the MRI tracking ability, this probe might enhance therapeutic immune activation as well.