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Adenoviral Delivery of Recombinant Soluble Human Tumor Necrosis Factor Receptor 1 Partially Normalized Mouse Model of Asthma
  1. Guo-Hua Huang, MD*,
  2. Xiao-Li Zeng, MD*,
  3. Yuan-Xiong Cheng, PhD*,
  4. Shun-Fang Zhu, MD*,
  5. Wei Luo, PhD,
  6. Qian Wen, PhD,
  7. Shao-Xi Cai, PhD*,
  8. Jin Su, PhD*
  1. From the *Department of Respiratory Diseases, Nanfang Hospital, and †Institute of Molecular Immunology, School of Biotechnology, Southern Medical University, Guangzhou, China.
  1. Received June 13, 2014, and in revised form February 25, 2015.
  2. Accepted for publication March 23, 2015.
  3. Reprints: Jin Su, Department of Respiratory Diseases, Nanfang Hospital, Southern Medical University, #1838, Northern Guangzhou Ave., Guangzhou, Guangdong, People’s Republic of China. E-mail: drsujin{at}126.com; or Shao-Xi Cai, Department of Respiratory Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. E-mail: hxkcai{at}126.com.
  4. This work was supported by the grants from the Guangdong province Medical research Foundation (A2011364), Nation Science Foundation (China) (81071847), and Guangdong Province Science Foundation (S2011010003881) and Guangdong Province Science Program (2012B031800394).
  5. The authors declare no conflict of interest.
  6. G.-H.H. and X.-L.Z. contributed equally to this work.

Abstract

Background Tumor necrosis factor α (TNF-α) is a proinflammatory cytokine that has been implicated in the airway pathology of asthma and result in resistance to hormone therapy. Tumor necrosis factor α inhibitors have become a major research focus in the treatment of asthma.

Methods Recombinant adenovirus (Ad-sTNFR1-IgGFc) expressing a fusion protein (sTNFR1-IgGFc), which was consisted of the soluble extracellular region of TNF receptor 1 and Fc fragment of IgG (sTNFR1-IgGFc), was used to transduce primary human airway smooth muscle cells (HASMCs). Enzyme-linked immunosorbent assay, flow cytometry, and immunocytochemistry confirmed the expression of sTNFR1-IgGFc. MTT was used to test the effect of sTNFR1-IgGFc to antagonism TNF-α–induced proliferates of HASMCs. To investigate the in vivo effectiveness of sTNFR1-IgGFc, mouse model of asthma was established. Ad-sTNFR1-IgGFc was delivered to the lung via nasal spray. Expression of sTNFR1-IgGFc in the tissue was confirmed by in situ hybridization and immunohistochemistry. The 2 major cell types that are involved in the inflamed asthmatic airway, neutrophils and eosinophils, in bronchoalveolar lavage fluid were observed.

Results The sTNFR1-IgGFc isolated from transduced HASMC culture supernatant was able to antagonize HASMC proliferation stimulated by TNF-α. Asthma-induced pathologies and alterations in the cell composition in bronchoalveolar lavage fluid were reduced in mice subjected to Ad-sTNFR1-IgGFc therapy.

Conclusions The soluble extracellular region of TNF receptor 1 and Fc fragment of IgG was able to functionally antagonize TNF-α in vitro and showed promise as a therapeutic agent for the localized treatment of severe refractory asthma.

Key Words
  • adenoviral vector
  • asthma
  • gene therapy
  • human airway smooth muscle cells (HASMCs)
  • soluble tumor necrosis factor α receptor (sTNFR)

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