title: In vivo Tracking of Dendritic Cell using MRI Reporter Gene, Ferritin.
abstract: The noninvasive imaging of dendritic cells (DCs) migrated into lymph nodes (LNs) can provide helpful information on designing DCs-based immunotherapeutic strategies. This study is to investigate the influence of transduction of human ferritin heavy chain (FTH) and green fluorescence protein (GFP) genes on inherent properties of DCs, and the feasibility of FTH as a magnetic resonance imaging (MRI) reporter gene to track DCs migration into LNs. FTH-DCs were established by the introduction of FTH and GFP genes into the DC cell line (DC2.4) using lentivirus. The changes in the rate of MRI signal decay (R2*) resulting from FTH transduction were analyzed in cell phantoms as well as popliteal LN of mice after subcutaneous injection of those cells into hind limb foot pad by using a multiple gradient echo sequence on a 9.4 T MR scanner. The transduction of FTH and GFP did not influence the proliferation and migration abilities of DCs. The expression of co-stimulatory molecules (CD40, CD80 and CD86) in FTH-DCs was similar to that of DCs. FTH-DCs exhibited increased iron storage capacity, and displayed a significantly higher transverse relaxation rate (R2*) as compared to DCs in phantom. LNs with FTH-DCs exhibited negative contrast, leading to a high R2* in both in vivo and ex vivo T2*-weighted images compared to DCs. On histological analysis FTH-DCs migrated to the subcapsular sinus and the T cell zone of LN, where they highly expressed CD25 to bind and stimulate T cells. Our study addresses the feasibility of FTH as an MRI reporter gene to track DCs migration into LNs without alteration of their inherent properties. This study suggests that FTH-based MRI could be a useful technique to longitudinally monitor DCs and evaluate the therapeutic efficacy of DC-based vaccines.
authors.affiliation: Department of Radiology, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul, Korea.
authors.forename: Hoe Suk
authors.initials: HS
authors.lastname: Kim
authors.affiliation: Department of Radiology, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul, Korea.
authors.forename: Jisu
authors.initials: J
authors.lastname: Woo
authors.affiliation: Department of Radiology, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul, Korea.
authors.forename: Jae Hoon
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Radiology, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul, Korea.
authors.forename: Hyun Jung
authors.initials: HJ
authors.lastname: Joo
authors.affiliation: Department of Radiology, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul, Korea; Department of Biomedical Science, College of Medicine, Seoul National University, 103 Daehak-ro, Jongno-gu, Seoul, Korea.
authors.forename: YoonSeok
authors.initials: Y
authors.lastname: Choi
authors.affiliation: Department of Radiology, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul, Korea; Department of Biomedical Science, College of Medicine, Seoul National University, 103 Daehak-ro, Jongno-gu, Seoul, Korea.
authors.forename: Hyeonjin
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Radiology, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul, Korea; Department of Biomedical Science, College of Medicine, Seoul National University, 103 Daehak-ro, Jongno-gu, Seoul, Korea.
authors.forename: Woo Kyung
authors.initials: WK
authors.lastname: Moon
authors.affiliation: Department of Radiology, Seoul Metropolitan Government Seoul National University, Boramae Medical Center, 20 Boramae-ro, Dongjag-gu, Seoul, Korea.
authors.forename: Seung Ja
authors.initials: SJ
authors.lastname: Kim
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D049109:Cell Proliferation / Q000502:physiology; D003713:Dendritic Cells; D005293:Ferritins / Q000235:genetics / Q000378:metabolism*; D005295:Ferrocyanides; D005434:Flow Cytometry; D017930:Genes, Reporter / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism; D016086:Lentivirus; D008198:Lymph Nodes / Q000276:immunology; D008279:Magnetic Resonance Imaging / Q000379:methods*; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D005295:Ferrocyanides; D053645:Interleukin-2 Receptor alpha Subunit; D013778:Tetrazolium Salts; D013844:Thiazoles; D049452:Green Fluorescent Proteins; D005293:Ferritins; C022616:thiazolyl blue; C000170:ferric ferrocyanide