Peng-Peng Zhang1, Xiao-Yong Xu, Feng Zhang, Yi Shi. 1. Department of Respiratory Medicine, Nanjing University/Nanjing General Hospital of Nanjing Military Command, PLA, Nanjing 210002, China.
Abstract
OBJECTIVE: To observe the changes of TLR-2, Dectin-1 expression on endothelial cells, and to explore their role in the immune response after contact with Aspergillus fumigatus. METHODS: Aspergillus fumigatus and human umbilical vein endothelial cells were co-incubated. Cells were collected respectively after incubation for 0 h, 1 h, 2 h, 4 h and 6 h. TLR2 and Dectin-1 receptor expressions were detected by flow cytometry, and their protein was measured by Western blot. The distribution of the receptors in the cells were observed by immunofluorescence. RESULT: TLR2 and Dectin-1 were expressed on the endothelial cell surface in quiescent condition. The mean fluorescence intensity of TLR2 on endothelial cells decreased from 45 to 13 stimulation by Aspergillus fumigatus, but the mean fluorescence intensity of Dectin-1 increased from 13 to 35 in the first 2 hours and then decreased. By Western blot, the electrophoresis strip of Dectin-1 was most bright in 2 hours after contact with the fungus, and then decreased 4 and 6 hours. TLR2 did not change significantly. Dectin-1 with fluorescent labeling was seen in spores and hyphae as well as in the cell membrane under confocal microscope. TLR2 was detected only on cell surface. CONCLUSION: TLR2 and Dectin-1 were expressed by endothelial cells, and may be useful in the identification of Aspergillus fumigatus.
OBJECTIVE: To observe the changes of TLR-2, Dectin-1 expression on endothelial cells, and to explore their role in the immune response after contact with Aspergillus fumigatus. METHODS:Aspergillus fumigatus and human umbilical vein endothelial cells were co-incubated. Cells were collected respectively after incubation for 0 h, 1 h, 2 h, 4 h and 6 h. TLR2 and Dectin-1 receptor expressions were detected by flow cytometry, and their protein was measured by Western blot. The distribution of the receptors in the cells were observed by immunofluorescence. RESULT: TLR2 and Dectin-1 were expressed on the endothelial cell surface in quiescent condition. The mean fluorescence intensity of TLR2 on endothelial cells decreased from 45 to 13 stimulation by Aspergillus fumigatus, but the mean fluorescence intensity of Dectin-1 increased from 13 to 35 in the first 2 hours and then decreased. By Western blot, the electrophoresis strip of Dectin-1 was most bright in 2 hours after contact with the fungus, and then decreased 4 and 6 hours. TLR2 did not change significantly. Dectin-1 with fluorescent labeling was seen in spores and hyphae as well as in the cell membrane under confocal microscope. TLR2 was detected only on cell surface. CONCLUSION:TLR2 and Dectin-1 were expressed by endothelial cells, and may be useful in the identification of Aspergillus fumigatus.