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[Detection of the expression of alpha3-integrin on hantavirus permissive cells]
Jingfang Dong1, Yuqing Jiang, Jianwei Wang
1Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052, China.
Background:
To express and purify human alpha3-integrin to serve as the antigen to prepare its antibody and to separate the Vero cell clones without expression of alpha3-integrin.
Methods:
The human alpha3-integrin gene was amplified by using RT-PCR, then subcloned into a pQE30 expression vector and expressed in E. coli. The gene expression was confirmed by Western blot assay. Rabbit was inoculated with purified antigen to stimulate the antibody generation. The target Vero cells were separated by negative selection using antibody plus complement mediated cytolysis. The separated cell clones were confirmed by immunofluorescence and Western blot assay.
Results:
The alpha3- integrin gene was cloned and expressed effetively, Western blot assay revealed that the expressed protein held good immune reactivity. High titer antibody was generated. However the expression of alpha3-integrin was not detected on Vero, VeroE6, Hep-2, 2BS and 293 cells.
Conclusions:
The results of the study suggested that hantavirus has other receptors on Vero cells beside alpha 3-integrin.