Pneumocystis carinii activates the NF-kappaB signaling pathway in alveolar epithelial cells.
Pneumocystis carinii pneumonia (PcP) is a clinically important infection of immunocompromised patients. Although the interaction of Pneumocystis with the alveolar epithelium has been well documented, very little information regarding the epithelial response to Pneumocystis is currently available. In order to study Pneumocystis-epithelium interactions, a murine cell line derived specifically from an alveolar epithelial cell (AEC) was utilized. The coculture of murine AECs with mouse Pneumocystis induced a dose- and time-dependent release of the CXC chemokine MIP-2. Importantly, the specific removal of Pneumocystis from the preparation, or the pretreatment of AECs with sulfasalazine, a potent and specific inhibitor of NF-kappaB, nearly completely abrogated the chemokine response to Pneumocystis. Since the murine MIP-2 promoter contains consensus kappaB binding sequences, the ability of Pneumocystis to stimulate NF-kappaB signaling in AECs was examined. Pneumocystis stimulation of an AEC line stably transfected with a kappaB-dependent reporter construct triggered the NF-kappaB signaling pathway and reporter production. These data were confirmed in gel shift assays, providing direct evidence that Pneumocystis induced the nuclear translocation of the p50/p65 heterodimeric form of NF-kappaB. Maximal NF-kappaB activation was dependent upon direct contact with viable Pneumocystis organisms. These data demonstrate that Pneumocystis activates NF-kappaB signaling in AECs and establish a reporter cell line for studying NF-kappaB activation in AECs. Given the global regulatory functions of the NF-kappaB family, these findings suggest that Pneumocystis directly alters AEC gene expression in a manner that promotes pulmonary immune and inflammatory responses.
Wang
Jing
J
Department of Pediatrics, P.O. Box 850, University of Rochester School of Medicine and Dentistry, 601 Elmwood Ave., Rochester, NY 14642, USA.
Gigliotti
Francis
F
Maggirwar
Sanjay
S
Johnston
Carl
C
Finkelstein
Jacob N
JN
Wright
Terry W
TW
D000818:Animals; D045744:Cell Line, Tumor; D054426:Chemokine CXCL2; D018925:Chemokines / Q000235:genetics / Q000378:metabolism*; D018920:Coculture Techniques; D004847:Epithelial Cells / Q000276:immunology / Q000382:microbiology*; D005260:Female; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D045363:Pneumocystis carinii / Q000276:immunology / Q000472:pathogenicity*; D011650:Pulmonary Alveoli / Q000166:cytology / Q000276:immunology / Q000382:microbiology*; D015398:Signal Transduction
D054426:Chemokine CXCL2; D018925:Chemokines; C404915:Cxcl2 protein, mouse; D016328:NF-kappa B