title: Successful use of mRNA-nucleofection for overexpression of interleukin-10 in murine monocytes/macrophages for anti-inflammatory therapy in a murine model of autoimmune myocarditis.
abstract: BACKGROUND
  Overexpression of interleukin-10 (IL-10) in murine CD11b(+) monocytes/macrophages via GMP-adapted mRNA-nucleofection was expected to improve clinical outcome and reduce adverse side effects in autoimmune myocarditis. This study represents the proof of principle for a novel anti-inflammatory therapy using overexpression of IL-10 in murine monocytes/macrophages by mRNA-nucleofection for the treatment of autoimmune myocarditis.
  
  
  RESULTS
  Autoimmune myocarditis was induced in A/J mice by subcutaneous immunization with troponin I. CD11b(+) monocytes/macrophages were isolated from the peritoneum and IL-10 was overexpressed by mRNA-nucleofection. These cells were injected intravenously. Myocardial inflammation was assessed via histological and immunohistochemical examinations. Myocardial fibrosis was analyzed with Masson's trichrome staining. Antitroponin I antibodies were determined within the serum. Physical performance was evaluated using a running wheel and echocardiography. In vitro overexpression of IL-10 in CD11b(+) monocytes/macrophages resulted in a 7-fold increased production of IL-10 (n=3). In vivo higher levels of IL-10 and less inflammation were detected within the myocardium of treated compared with control mice (n=4). IL-10-treated mice showed lower antitroponin I antibodies (n=10) and a better physical performance (n=10).
  
  
  CONCLUSIONS
  Application of IL-10-overexpressing CD11b(+) monocytes/macrophages reduced inflammation and improved physical performance in a murine model of autoimmune myocarditis. Thus, the use of genetically modified monocytes/macrophages facilitated a targeted therapy of local inflammation and may reduce systemic side effects. Because the nucleofection technique is GMP adapted, an in vivo use in humans seems basically feasible and the transfer to other inflammatory diseases seems likely.
authors: Cardiovascular Center Oberallgäu, Kempten, Germany.
authors: Oliver
authors: O
authors: Zimmermann
authors: Jörg M
authors: JM
authors: Homann
authors: Anna
authors: A
authors: Bangert
authors: Anna-Maria
authors: AM
authors: Müller
authors: Georgi
authors: G
authors: Hristov
authors: Stefan
authors: S
authors: Goeser
authors: Juliane M
authors: JM
authors: Wiehe
authors: Stefan
authors: S
authors: Zittrich
authors: Wolfgang
authors: W
authors: Rottbauer
authors: Jan
authors: J
authors: Torzewski
authors: Gabriele
authors: G
authors: Pfitzer
authors: Hugo A
authors: HA
authors: Katus
authors: Ziya
authors: Z
authors: Kaya
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000097:blood / Q000378:metabolism / Q000627:therapeutic use*; D001327:Autoimmune Diseases / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy*; D002465:Cell Movement; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005240:Feasibility Studies; D005260:Female; D015870:Gene Expression; D006801:Humans; D007249:Inflammation / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy; D016753:Interleukin-10 / Q000097:blood / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D008264:Macrophages / Q000378:metabolism / Q000502:physiology; D051379:Mice; D009000:Monocytes / Q000378:metabolism / Q000502:physiology; D009205:Myocarditis / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy*; D012333:RNA, Messenger; D014162:Transfection
chemical_list: D000893:Anti-Inflammatory Agents; D012333:RNA, Messenger; D016753:Interleukin-10
keywords: cell therapy; heart failure; inflammation; interleukin-10; myocarditis