Construction of aquaporin-9 expressing vector and detection of AQP-9 antibody in patients with neuromyelitis optica
GAO Cong
ZHENG Yang-bo
SHAN Fu-lan
FAN Yong-xiang
WU Lin-zhan
ZHONG Rong
LONG You-ming
Abstract：Objective To construct a eukaryotic aquaporin -9 (AQP9) expressing vector for AQP9 antibody de-tection in patients with neuromyelitis optica (NMO).Methods The recombinant vector was constructed by subcloning human AQP9 cDNA into pcDNA3.1+vector, with which human embryonic kidney (HEK) 293T cells was transfected to establish an AQP9 protein expressing cell line .The cell line expression AQP 9 was confirmed by reverse transcription poly-merase chain reaction ( RT-PCR) and Western blot analysis .Serum antibody was tested with cell -based assay by indi-rect immunofluorescence assay .Results The relative expression level of AQP 9 mRNA was significantly higher than the empty vector, and Western blotting analysis showed the expression content at 32 kD bands was significantly higher than the empty vector and 293T cells.The cell line expressing AQP9 was successfully constructed and confirmed by reverse tran-scription polymerase chain reaction ( RT-PCR) and Western blot analysis .The serum antibody was positive in all 4 pa-tients with NMO and negative in one with cerebral infarction and one normal adult .Conclusion The transfected cell mod-el is simple and advantageous in AQP 9 serum antibody detection for research on NMO .
Keywords：Aquaporin-9neuromyelitis opticaHEK 293Tantibody
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 821-824 )