GM-CSF regulates a PU.1-dependent transcriptional program determining the pulmonary response to LPS

被引:65
作者
Berclaz, Pierre-Yves
Carey, Brenna
Fillipi, Marie-Dominique
Wernke-Dollries, Kara
Geraci, Nick
Cush, Stephanie
Richardson, Terry
Kitzmiller, Joe
O'Connor, Michael
Hermoyian, Christina
Korfhagen, Thomas
Whitsett, Jeffrey A.
Trapnell, Bruce C.
机构
[1] Childrens Hosp, Med Ctr, Div Pulm Biol, Cincinnati, OH 45229 USA
[2] Childrens Hosp, Med Ctr, Div Expt Hematol, Cincinnati, OH 45229 USA
[3] Univ Cincinnati, Coll Med, Cincinnati, OH USA
关键词
alveolar macrophages; inflammation; lung; neutrophils;
D O I
10.1165/rcmb.2006-0174OC
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Alveolar macrophages (AMs) normally respond to lipopolysaccharide (LIPS) by activating Toll-like receptor (TLR)-4 signaling, a mechanism critical to lung host defense against gram-negative bacteria such as Pseudomonas aeruginosa. Because granulocyte macrophage colony-stimulating factor (GM-CSF)-deficient (GM(-/-)) mice are hyporesponsive to LPS, we evaluated the role of GM-CSF in TLR-4 signaling in AMs. Pulmonary TNF-alpha levels and neutrophil recruitment 4 h after intratracheal administration of Pseudomonas LIPS were reduced in GM(-/-) compared with wild-type (G(+/+)) mice. Secretion of TNF-alpha by AMs exposed to LPS ex vivo was also reduced in GM(-/-) mice and restored in mice expressing GM-CSF specifically in the lungs (SPC-GM(+/+)/GM(-/-) mice). LPS-dependent NF-kappa B promoter activity, TNF-alpha secretion, and neutrophil chemokine release were reduced in AM cell lines derived from GM(-/-) mice (mAM) compared with GM(+/+) (MH-S). Retroviral expression of PU.1 in mAM cells, which normally lack PU.1, rescued all of these AM defects. To determine whether GM-CSF, via PU.1, regulated expression of TLR-4 pathway components, mRNA and protein levels for key components were evaluated in MH-S cells (GM(+/+), PU.1(Positive)), mAM cells (GM(-/-), PU.1(Negative)), and mAMPU.1+ cells (GM(-/-), PU.1(Positive)). Cluster of differentiation antigen-14, radioprotective 105, IL-1 receptor-associated kinase (IRAK)-M mRNA, and protein were dependent upon GM-CSF and restored by expression of PU.1. In contrast, expression of other TLR-4 pathway components (myeloid differentiation-2, TLR-4, IRAK-1, IRAK-2, Toll/IL-1 receptor domain containing adapter protein/MyD88 adaptor-like, myeloid differentiation primary-response protein 88, IRAK-4, TNF receptor-associated factor-6, NF-kappa B, inhibitor of NF-kappa B kinase) were not GM-CSF or PU.1-dependent. These results show that GM-CSF, via PU.1, enables AM responses to P. aeruginosa LIPS by regulating expression of a specific subset of components of the TLR-4 signaling pathway.
引用
收藏
页码:114 / 121
页数:8
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