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(American Journal of Pathology. 2005;167:1321-1331.)
© 2005 American Society for Investigative Pathology

Ex Vivo Expanded Dendritic Cells Home to T-Cell Zones of Lymphoid Organs and Survive in Vivo after Allogeneic Bone Marrow Transplantation

Christoph H. Schimmelpfennig*{dagger}, Stephan Schulz*{ddagger}, Caroline Arber*, Jeanette Baker*, Ingo Tarner§, Jacqueline McBride§, Christopher H. Contag and Robert S. Negrin*

From the Divisions of Blood and Marrow Transplantation* and Immunology and Rheumatology,§ Department of Medicine, and the Department of Pediatrics, Stanford University, Stanford, California; the Department of Medicine,{dagger} Knappschaftskrankenhaus, Ruhr-Universitaet Bochum, Bochum, Germany; and the Institute of Pathology,{ddagger} Klinikum Rechts der Isar, Technical University of Munich, Munich, Germany

Little is known about adoptive transfer of allogeneic ex vivo expanded dendritic cells (eDCs). We investigated the trafficking pattern of eDCs in mice after allogeneic bone marrow transplantation by using bioluminescence imaging. eDCs were expanded from bone marrow precursors in the presence of GM-CSF, interleukin-4, and Flt3L and retrovirally transduced to express luciferase (luc) and green fluorescence protein (gfp). Flow cytometry showed polyclonal DC populations after expansion that consisted of CD11c+CD11b+ and CD11c–CD11b+ cells that co-expressed CD40, CD80, CD86, and MHCII. eDCs were functional in mixed lymphocyte reactions and produced tumor necrosis factor-{alpha} on phytohemagglutinin stimulation. The eDCs were then injected intravenously into BALB/c recipient mice that had received allogeneic bone marrow transplantation 6 weeks previously. On day 1 after transfer, eDCs were detected by bioluminescence imaging throughout the lungs and spleen. In the later course, signals were observed throughout thymus, lower abdomen, and spleen throughout a period of more than 42 days. Immunofluorescence microscopy confirmed CD11c positivity on the gfp+ donor cells, which localized in T-cell zones of mesenteric lymph nodes, Peyer’s patches, spleen, and thymus. These findings are important for adoptive immunotherapies because they indicate that eDCs migrate efficiently in vivo and are capable of surviving long term.




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