Mucosal delivery of a double-stapled RSV peptide prevents nasopulmonary infection
Gregory H. Bird, … , Shyam S. Mohapatra, Loren D. Walensky
Gregory H. Bird, … , Shyam S. Mohapatra, Loren D. Walensky
Published April 17, 2014
Citation Information: J Clin Invest. 2014;124(5):2113-2124. https://doi.org/10.1172/JCI71856.
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Research Article

Mucosal delivery of a double-stapled RSV peptide prevents nasopulmonary infection

Abstract

Respiratory syncytial virus (RSV) infection accounts for approximately 64 million cases of respiratory disease and 200,000 deaths worldwide each year, yet no broadly effective prophylactic or treatment regimen is available. RSV deploys paired, self-associating, heptad repeat domains of its fusion protein, RSV-F, to form a fusogenic 6-helix bundle that enables the virus to penetrate the host cell membrane. Here, we developed hydrocarbon double-stapled RSV fusion peptides that exhibit stabilized α-helical structure and striking proteolytic resistance. Pretreatment with double-stapled RSV peptides that specifically bound to the RSV fusion bundle inhibited infection by both laboratory and clinical RSV isolates in cells and murine infection models. Intranasal delivery of a lead double-stapled RSV peptide effectively prevented viral infection of the nares. A chitosan-based nanoparticle preparation markedly enhanced pulmonary delivery, further preventing progression of RSV infection to the lung. Thus, our results provide a strategy for inhibiting RSV infection by mucosal and endotracheal delivery of double-stapled RSV fusion peptides.

Authors

Gregory H. Bird, Sandhya Boyapalle, Terianne Wong, Kwadwo Opoku-Nsiah, Raminder Bedi, W. Christian Crannell, Alisa F. Perry, Huy Nguyen, Viviana Sampayo, Ankita Devareddy, Subhra Mohapatra, Shyam S. Mohapatra, Loren D. Walensky

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Figure 4

Inhibition of nasal RSV infection by intranasal pretreatment with SAH-RSVFBD.

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Inhibition of nasal RSV infection by intranasal pretreatment with SAH-RS...
(A) 10-week-old BALB/c mice were treated with vehicle (1.2% DMSO) or the indicated SAH-RSVF peptides (50 μl of 125 μM solution in 1.2% DMSO) by nasal drop, followed by inoculation with rgRSV (1 × 106 pfu/mouse). Tissues were harvested 24 hours after infection and processed for fluorescence imaging, which revealed marked suppression of GFP positivity by SAH-RSVFBD, but not the control SAH-RSVFBF. Original magnification, ×20. (B) Average GFP positivity, quantified by ImageJ analysis of 8 images (4 sections) per mouse. Data (mean ± SEM) represent percent GFP+ of total DAPI+ cells. (C) H&E staining of the nasal sections demonstrated airway constriction from mucous production (a disease hallmark in mice; yellow arrow) and inflammation (mononuclear cell infiltrates; white arrow) in rgRSV-infected mice treated with vehicle and SAH-RSVFBF, but notably less airway constriction and immune cell infiltration in SAH-RSVFBD–treated mice. Sections from mock-infected mice served as a negative control for nasal mucosal inflammation. Insets show magnified views of yellow boxed regions. Original magnification, ×20; ×40 (insets). *P < 0.001.