Ag85B lentivirus inhibits expression of TSLP and TSLP receptor in human bronchial epithelial cells |
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Authors: | FANG Xiang-feng QIAN Jiang MENG Jin-xiu LI Dong-feng WU Jian |
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Affiliation: | 1.Postgraduate Institute of Southern Medical University, Guangzhou 510515, China;2.Department of Respiratory Medicine, 3Medical Research Center, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China. |
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Abstract: | AIM: To investigate the effect of Mycobacterium tubeculosisantigen (Ag85B) lentivirus on the expression of thymic stromal lymphopoietin (TSLP) and TSLP receptor (TSLPR) in cultured normal human bronchial epithelial cells (NHBEC) in vitro. METHODS: Ag85Bgene was subcloned into 293T cells with recombinant plasmid to yield the Ag85B lentivirus. The NHBEC were cultured and transfected with Ag85B lentivirus, NS control and empty lentivirus control using Lipofectamine 2000. The expression of Ag85B, TSLP and TSLPR in NHBEC at mRNA and protein levels was confirmed by real-time PCR and Western blotting.RESULTS: Ag85B lentivirus was constructed with a viral titer of 2×1011 TU/L and the Ag85B expression was detected. As compared with control group and empty lentivirus group, the mRNA expression of TSLP in Ag85B lentivirus group was found to slightly decreased, and the mRNA expression of TSLPR and the protein expression of TSLP and TSLPR were significantly decreased in Ag85B lentivirus group (P<0.05). CONCLUSION: The constructed Ag85B lentivirus antagonizes the expression of TSLP and TSLPR in cultured NHBEC, suggesting that Ag85B inhibits the initiation of asthmatic inflammation by blockage of TSLP-TSLPR pathway. |
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Keywords: | Thymic stromal lymphopoietin Thymic stromal lymphopoietin receptor Lentiviral vector Antigen 85B Human bronchial epithelia cells |
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