Recombinant porcine lung surfactant protein A inhibits porcine reproductive and respiratory syndrome virus infection into host cells in vitro].

To investigate the antiviral activity of porcine lung surfactant protein A (SP-A) to porcine reproductive and respiratory syndrome virus (PRRSV) in vitro. The SP-A gene was amplified by PCR from the plasmid containing porcine SP-A gene, and subcloned into pcDNA3. 1A-CD5 vector containing the human CD5 signal peptide to generate SP-A eukaryotic expression vector pcDNA-CD5-SPA/MH. The recombinant expression vector was transfected into HEK293T cells mediated with calcium phosphate. The expressed recombinant SP-A was identified by Western blot and purified from culture medium by Ni-NTA-Agarose beads. The binding activity of SP-A with PRRSV was identified by ELISA. The antiviral activity of SP-A to PRRSV was analyzed by viral titer reduction assays on MARC-145 cells and porcine alveolar macrophages (PAM). The results showed that the eukaryotic expression vector of SP-A gene could mediate SP-A expression in HEK293T cells, the expressed SP-A could bind PRRSV in a dose dependent manner. The PRRSV incubated in advance with SP-A showed the lower infective activity compared with no-SP-A-incubated PRRSV on both MARC-145 cells and porcine alveolar macrophages. The SP-A-treated PRRSV titers in MARC-145 cells and PAM cells were significantly lower than that of SP-A-untreated PRRSV at 72 h post-infection. Recombinant porcine SP-A significantly inhibit the infection of PRRSV to the host cells in vitro, which indicates that recombinant SP-A possesses anti-PRRSV activity.

Citation

Feng Zhang, Fei Zhong, Xiujin Li, Xingxing Wang, Kao Zhang, Huihui Chen, Zhen Li, Wenyan Li, Hongli Pan, Dongmei Han. Recombinant porcine lung surfactant protein A inhibits porcine reproductive and respiratory syndrome virus infection into host cells in vitro]. Wei sheng wu xue bao = Acta microbiologica Sinica. 2012 May 4;52(5):654-60

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PMID: 22803352

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