Lentiviral vector-mediated stable expression of sTNFR-Fc in human macrophage and neuronal cells as a potential therapy for neuroAIDS

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• 作者：Shengbo Cao (1) (3)
  Chengxiang Wu (1)
  Yongbo Yang (1)
  Lynn F Sniderhan (2)
  Sanjay B Maggirwar (2)
  Stephen Dewhurst (2)
  Yuanan Lu (1)
  
• 刊名：Journal of Neuroinflammation
• 出版年：2011
• 出版时间：December 2011
• 年：2011
• 卷：8
• 期：1
• 全文大小：2060KB
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• 作者单位：Shengbo Cao (1) (3)
  Chengxiang Wu (1)
  Yongbo Yang (1)
  Lynn F Sniderhan (2)
  Sanjay B Maggirwar (2)
  Stephen Dewhurst (2)
  Yuanan Lu (1)
  
  1. Department of Public Health Sciences, University of Hawai’i, Honolulu, Hawai’i, 96822, USA
  3. College of Veterinary Medicine, Huazhong Agricultural University, Hubei, Wuhan, 430070, China
  2. Department of Microbiology & Immunology, University of Rochester, Rochester, NY, 14642, USA
  
• ISSN：1742-2094

文摘

Background Human immunodeficiency virus type 1 (HIV-1) infection frequently causes neurologic disease, which is the result of viral replication and activation of macrophages and microglia in the CNS, and subsequent secretion of high levels of neurotoxic products, including tumor necrosis factor-α (TNF-α). We therefore hypothesized that a soluble TNF-α antagonist might have potential utility as a neuroprotective effecter molecule, and conducted proof-of-concept studies to test this hypothesis. Methods To develop novel therapeutics for the treatment of neuroAIDS, we constructed and characterized a soluble TNF receptor (sTNFR)-Fc fusion protein with the goal of neutralizing TNF-α, and tested the stability of expression of this gene following delivery by a lentiviral vector. Results High-titer lentiviral vectors were prepared, allowing efficient transduction of macrophage/glial and neuronal cell lines, as well as primary rat cerebellar neurons. Efficient, stable secretion of sTNFR-Fc was demonstrated in supernatants from transduced cell lines over 20 passages, using both western blot and ELISA. Biological activity of the secreted sTNFR-Fc was confirmed by TNF-specific in vitro protein binding and functional blocking assays. Finally, the secreted protein was shown to protect neuronal cells from TNF-α, HIV-1 Tat-, and gp120-mediated neurotoxicity. Conclusions These results demonstrate that lentiviral vector mediated expression of sTNFR-Fc may have potential as a novel therapy for neuroAIDS.