Detection and characterization of OX40 ligand expression in human airway smooth muscle cells: a possible role in asthma?
BACKGROUND
The airway smooth muscle (ASM) cell, originally thought of as a passive structural cell, is now well recognized as an active participant in the pathologic events that occur during persistent asthma. Cell-surface molecules play an important role in the development of an immune response. A number of cell-surface molecules are expressed on ASM cells, and these might contribute to the inflammatory reaction.


OBJECTIVE
The purpose of this study was to determine whether OX40 ligand (OX40L), a molecule known to be involved in T-cell activation, was present on the ASM cell surface.


METHODS
We used real-time RT-PCR to detect mRNA expression and flow cytometry, ELISA, and immunoprecipitation to detect the presence of cell-surface protein on ASM cells isolated from asthmatic and nonasthmatic individuals. ELISAs and Western blotting were used to determine the functional outcomes of engagement of OX40L.


RESULTS
OX40L was present on both asthmatic and nonasthmatic ASM cells. Engagement of OX40L with recombinant OX40:Fc resulted in a significantly greater increase in release of IL-6 from ASM cells of asthmatic patients than from ASM cells of nonasthmatic patients (P<.01). Ligation of OX40L resulted in a rapid translocation of protein kinase C beta2 to the cell membrane.


CONCLUSIONS
Because the receptor for OX40L, OX40, is expressed on CD4+ T cells within 48 hours of stimulation through the T-cell receptor, elucidation of the cross-talk between OX40 and OX40L could be very important in understanding the interaction of cells present in the inflamed airways of an asthmatic patient.
Burgess
Janette K
JK
Respiratory Research Group, Department of Pharmacology, Bosch Building D05, University of Sydney, Sydney, NSW 2006, Australia.
Carlin
Stephen
S
Pack
Robert A
RA
Arndt
Greg M
GM
Au
Wendy W
WW
Johnson
Peter R A
PR
Black
Judith L
JL
Hunt
Nicholas H
NH
D000293:Adolescent; D000328:Adult; D000368:Aged; D000943:Antigens, Differentiation / Q000494:pharmacology; D001249:Asthma / Q000503:physiopathology; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008875:Middle Aged; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D053278:OX40 Ligand; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000378:metabolism; D012137:Respiratory System / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects
D000943:Antigens, Differentiation; D016207:Cytokines; D008562:Membrane Glycoproteins; D053278:OX40 Ligand; C406562:OX40Ig; D012333:RNA, Messenger; C089251:TNFSF4 protein, human