title: Nucleotide oligomerization domain 2 contributes to the innate immune response in THCE cells stimulated by Aspergillus fumigatus conidia.
abstract: OBJECTIVE
  To investigate the expression of nucleotide oligomerization domain 2 (NOD2) in the immortalized human corneal epithelial cell line (THCE), and its role in the innate immune response triggered by inactive Aspergillus fumigatus (Af) conidia.
  
  
  METHODS
  The normal THCE cells were investigated as controls. After incubation with inactive Af conidia for 0.5, 2, 4, 6, and 8 hours, THCE cells were harvested, mRNA expression of NOD2 and receptor interacting protein 2 (RIP2) was detected by RT-PCR. Intracellular proteins including NOD2, NF-κB and proinflammatory cytokines such as TNF-α, IL-8, IL-6 in the cell supernatant were analyzed by ELISA.
  
  
  RESULTS
  Our data indicate that NOD2 expressed in the normal THCE cells. After triggered by the inactive Af conidia, the expression of NOD2, RIP2 mRNA and the secretion of NOD2, NF-κB, TNF-α, IL-8, IL-6 both increased in a time-depended manner, and reached the peak point at 4, 6, 6, 4, 6, 6, 4 hours, respectively. And after pretreated with NOD2 neutralizing antibody, the expression of RIP2, NF-κB, TNF-α, IL-8 both decreased dramatically at the peak point, while the secretion of IL-6 changed little.
  
  
  CONCLUSIONS
  The results of this study suggest that NOD2 exists and expresses in the THCE cells, and contributes to the innate immune responses triggered by inactive Af conidia by induction of proinflammatory cytokines such as TNF-α and IL-8 through the NF-κB pathway.
authors.affiliation: Department of Ophthalmology, the Affiliated Hospital of Qingdao University Medical College, Qingdao 266003, Shandong Province, China.
authors.forename: Zheng-Jie
authors.initials: ZJ
authors.lastname: Xu
authors.forename: Gui-Qiu
authors.initials: GQ
authors.lastname: Zhao
authors.forename: Qing
authors.initials: Q
authors.lastname: Wang
authors.forename: Cheng-Ye
authors.initials: CY
authors.lastname: Che
authors.forename: Nan
authors.initials: N
authors.lastname: Jiang
authors.forename: Li-Ting
authors.initials: LT
authors.lastname: Hu
authors.forename: Qiang
authors.initials: Q
authors.lastname: Xu