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Published ahead of print on March 20, 2003, doi:10.1165/rcmb.2002-0069OC

Am. J. Respir. Cell Mol. Biol., Volume 29, Number 2, August 2003, 213-224

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Submitted on May 29, 2002
Revised on March 19, 2003

Bone Marrow Origin of Myofibroblasts in Irradiation Pulmonary Fibrosis

Michael W Epperly1, Hongliang Guo1, Joan E Gretton1, and Joel S Greenberger1*

1 Radiation Oncology, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA

* To whom correspondence should be addressed. E-mail: greenbergerjs{at}msx.upmc.edu.

There is a rapid onset of organizing alveolitis/fibrosis at 120-140 days after whole lung irradiation of C57BL/6J mice. To test the hypothesis that circulating cells of bone marrow origin contribute to irradiation fibrosis, irradiated chimeric green fluorescent protein (GFP)+ C57BL/6J mice were followed for GFP+ cells in areas of lung fibrosis. In a second experimental model, C57BL/6J female mice received 20 Gy total lung irradiation, and after 60 or 80 days were intravenously injected with cells from a clonal GFP+ male bone marrow stromal cell line or GFP+ whole bone marrow, respectively. The mice were then followed for the development of pulmonary fibrosis and the contribution of GFP+ cells to fibrotic areas was quantitated. Bromodeoxyuridine (BrdU) labeling of developing fibrotic areas showed that the cell division occurred predominantly in GFP+, Y-probe positive, and vimentin positive cells. Immunohistochemistry demonstrated that these cells were macrophages and fibroblasts, not endothelial cells. Mice that received manganese superoxide dismutase-plasmid/liposome (MnSOD-PL) intratracheal injection 24 hours prior to total lung irradiation demonstrated a decrease in GFP+ fibroblastic cells in the lung. Thus, pulmonary irradiation fibrosis contains proliferating cells of bone marrow origin, and gene therapy prevention of this condition acts in part by decreasing the migration and proliferation of marrow origin cells.




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