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Originally published online as doi:10.1189/jlb.0104037 on June 14, 2004

Published online before print June 14, 2004
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(Journal of Leukocyte Biology. 2004;76:472-476.)
© 2004 by Society for Leukocyte Biology

Analysis of the CCR7 expression on murine bone marrow-derived and spleen dendritic cells

Uwe Ritter*,1, Florian Wiede*, Dirk Mielenz{dagger}, Ziba Kiafard{ddagger}, Jörg Zwirner{ddagger} and Heinrich Körner*,§,2

* IZKF Nachwuchsgruppe 1, Nikolaus-Fiebiger Zentrum für Molekulare Medizin, and
{dagger} Abteilung für Immunologie, Nikolaus-Fiebiger Zentrum für Molekulare Medizin, Universität Erlangen, Germany;
{ddagger} Abteilung für Immunologie, Georg-Augustus-Universität, Göttingen, Germany; and
§ Comparative Genomics Centre, James Cook University, Townsville, Australia

1Correspondence: Nikolaus-Fiebiger-Zentrum für Molekulare Medizin, IZKF, Nachwuchsgruppe 1, Glückstrasse 6, D-91054 Erlangen, Germany. E-mail: uritter{at}molmed.uni-erlangen.de

About 40% of bone marrow-derived dendritic cells (BM-DCs) generated from stem cells of C57BL/6 (B6.WT) mice differentiate in the presence of granulocyte macrophage-colony stimulating factor (GM-CSF) without further stimuli to mature DCs. These cells are characterized by high levels of major histocompatibility complex class II, CD40, and CD86 on their surface. Recent studies have revealed that tumor necrosis factor (TNF) is crucial for maturation of BM-DCs. However, once matured, the phenotype of mature TNF-negative C57BL/6 (B6.TNF–/–) and B6.WT BM-DCs is comparable. Both expressed high levels of CD40 and CD86 and were positive for mRNA of the chemokine receptor (CCR)7. To extend our studies, we generated a monoclonal antibody (mAb) specific for mouse CCR7. This mAb allowed us to analyze the surface expression of CCR7 during maturation of B6.WT and B6.TNF–/– BM-DCs in the presence of GM-CSF and stimulated with TNF or lipopolysaccharide (LPS) and to compare it with the CCR7 expression on ex vivo-isolated splenic DCs with or without additional stimulation. Our results showed that CCR7 expression on murine BM-DCs is an indication of cell maturity. Incubation with LPS induced the maturation of all BM-DCs in culture but increased the number of mature CCR7+ splenic DCs only marginally.

Key Words: tumor necrosis factor • mouse • knockout • bone marrow precursor cells • chemokine receptor




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