Accuri C6 Flow Cytometer System
Originally published online as doi:10.1189/jlb.0606377 on October 19, 2006

Published online before print October 19, 2006
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(Journal of Leukocyte Biology. 2007;81:186-194.)
© 2007 by Society for Leukocyte Biology

Macrophages in experimental rat lung isografts and allografts: infiltration and proliferation in situ

Andree Schmidt*,1, Jochen Sucke*,1, Gabriele Fuchs-Moll*, Petra Freitag*, Markus Hirschburger*, Andreas Kaufmann*, Holger Garn{dagger}, Winfried Padberg* and Veronika Grau*,2

* Laboratory of Experimental Surgery, Department of General and Thoracic Surgery, Justus-Liebig-University Giessen, Giessen, Germany; and
{dagger} Department of Clinical Chemistry and Molecular Diagnostics, Hospital of the Philipps-University, Marburg, Germany

2Correspondence: Laboratory of Experimental Surgery, Department of General and Thoracic Surgery, Justus-Liebig-University Giessen, Rudolf-Buchheim-Str. 7, D-35385 Giessen, Germany. E-mail: veronika.grau{at}chiru.med.uni-giessen.de

ABSTRACT

Alveolar macrophages (AMs) and peribronchial/perivascular macrophages are probably involved in lung allograft damage. We investigate leukocyte infiltration into graft tissue and address the question whether proliferation in situ contributes to macrophage homeostasis and accumulation. Lung transplantation was performed in the Lewis (LEW)-to-LEW and in the Dark Agouti-to-LEW rat strain combination. Graft infiltration by ED1+ and ED2+ (CD163) macrophages was analyzed by immunohistochemistry (IHC) and compared with infiltration by lymphocytes. Cells in the S-phase of the cell cycle were pulse-labeled with BrdU and detected immunohistochemically. Finally, the donor or recipient origin of AMs was determined by IHC and in situ hybridization. ED1+ AMs in allogeneic transplants increased by more than 25-fold from Days 1 to 5. In addition, large, peribronchial/perivascular infiltrates developed containing numerous ED1+ cells. Although AMs in normal rat lungs are CD163–, AMs up-regulated CD163 between Days 4 and 5, reaching maximum values on Day 6. Lymphocytes were less numerous than macrophages. About 16% of the AMs and 10% of the peribronchial/perivascular macrophages were in the S-phase of the cell cycle on Day 2 post-transplantation. No differences in the frequency of BrdU+ macrophages were obvious between isografts and allografts. AMs of donor origin increased in number considerably during allograft rejection. In conclusion, the cellular infiltrate in lung allografts is dominated by macrophages, which exhibit an unusual phenotype and a strong capacity for mitotic self-renewal.

Key Words: lung transplantation • alveolar macrophage • acute rejection • CD163




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