RT Journal Article
SR Electronic
T1 Parenchymal cell populations involved in the development of fibrosis
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP P3759
VO 44
IS Suppl 58
A1 Birgitta Christine Heckl
A1 Isis Enlil Fernandez Buelvas
A1 Katharina Heinzelmann
A1 Oliver Eickelberg
YR 2014
UL http://erj.ersjournals.com/content/44/Suppl_58/P3759.abstract
AB Lung fibrosis is a severe, chronic lung disease, with a median survival of 3 – 5 years after diagnosis. While there has been significant progress in defining cell-specific functions in disease development and progression, a detailed analysis of definitive cell compartments during fibrosis remains to be elucidated.Intratracheal installation of bleomycin (3U/Kg) or PBS was performed in C57/bl6 mice, and single cell suspensions of the lungs were obtained at day 14 after treatment. To analyze dynamic changes in the parenchyma, epithelial, endothelial, and mesenchymal markers were used for flow cytometry analysis. Parenchymal CD45- cells were sorted and further analyzed by immunofluorescence staining for epithelial, endothelial cells and mesenchymal cells.FACS analysis of single cell suspensions from lung homogenates confirmed an increase in cellularity during the onset of fibrosis. Cell-specific analysis for mesenchymal, endothelial, and epithelial markers revealed an increase in CD45- cells at day 14 to 3.3x106 (144.75%), compared with 2.3x106 (100%) of controls. Epithelial cells were gated as CD45-, EpCAM+ and showed a slight increase during fibrosis (6.1x105 -132.46%- vs. 4.6x105 -100%-). Endothelial CD45-, CD31+ cells showed up to two-fold increase (2.8x106 -187.30%- vs. 1.5x106 -100%-), similar to the mesenchymal CD45-, CD90.2+ population (2.7x105 -274.59%- vs. 1.0x105 -100%-). Sorted CD45- CD31+ CD90.2 -cells showed a positive staining for T1α, E-Cadherin, proSPC and α-SMA.Our results show that an overall increase in cellularity is a critical determinant of parenchymal expansion in lung fibrosis. To characterize these populations more in detail, further analysis needs to be done.