Nasal polyp fibroblasts produce MIP-3alpha in response to toll-like receptor ligands and cytokine stimulation.
OBJECTIVE
Dendritic cells (DCs) play important roles in the development and perpetuation of immune responses. DCs are present in upper airway diseases such as chronic rhinosinusitis with nasal polyps. However, the mechanisms of how DCs migrate into the upper airway mucosa during upper airway inflammatory diseases remains unclear. Macrophage inflammatory protein-3alpha (MIP-3alpha) is known to be a migratory factor for immature DCs. There have been very few reports regarding cells producing this chemokine in the airways. To investigate this, we stimulated fibroblasts cultured from the nasal polyps with various toll-like receptor (TLR) ligands, which are derived from microorganisms, and IL-beta1 and TNF-alpha, which are proinflammatory cytokines, and analyzed their ability to produce MIP-3alpha.


METHODS
Fibroblast lines were established from nasal polyps and stimulated with TLR2, 3, 4, 5, 7/8 and 9 ligands, IL-beta1 and TNF-alpha. MIP-3alpha mRNA expression in nasal polyp fibroblasts (NPF) was evaluated by real-time RT-PCR and the protein levels of MIP-3alpha in the supernatants of stimulated NPF was measured by ELISA.


RESULTS
Stimulation with TLR2, 3, 4 and 5 ligands, IL-beta1 and TNF-alpha, induced MIP-3alpha gene expression and protein production in the cultured NPF This response was dose- and time-dependent.


CONCLUSIONS
NPF possibly play an important role in the recruitment of DCs in upper airway diseases such as chronic rhinosinusitis with nasal polyps through the production of MIP-3alpha.
Nonaka
Manabu
M
Department of Otolaryngology, Nippon Medical School, Sendagi, Tokyo, Japan. nonaka@nms.ac.jp
Ogihara
Nozomu
N
Fukumoto
Akira
A
Sakanushi
Atsuko
A
Kusama
Kaoru
K
Pawankar
Ruby
R
Yagi
Toshiaki
T
D054418:Chemokine CCL20 / Q000378:metabolism*; D003713:Dendritic Cells / Q000502:physiology; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D053583:Interleukin-1beta / Q000502:physiology; D008024:Ligands; D009298:Nasal Polyps / Q000378:metabolism* / Q000473:pathology; D051193:Toll-Like Receptors / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology
C108355:CCL20 protein, human; D054418:Chemokine CCL20; D053583:Interleukin-1beta; D008024:Ligands; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha