Faculty Collaboration Database

In vitro transport of active alpha(1)-antitrypsin to the apical surface of epithelia by targeting the polymeric immunoglobulin receptor. Am J Respir Cell Mol Biol 1999 Aug;21(2):246-52

Date

07/29/1999

Pubmed ID

10423408

DOI

10.1165/ajrcmb.21.2.3687

Scopus ID

2-s2.0-0033172972 (requires institutional sign-in at Scopus site)   15 Citations

Abstract

In cystic fibrosis (CF), the intense host inflammatory response to chronic infection largely accounts for the progressive pulmonary disease, and ultimately death. Neutrophils are the prominent inflammatory cells in the lungs of patients with CF, and large amounts of neutrophil elastase (NE) are released during phagocytosis. Besides having direct effects on structural elastin, NE stimulates the release of proinflammatory mediators from the respiratory epithelium and is a potent secretogogue. Therapeutic use of elastase inhibitors in CF has been complicated by difficulties in delivery to the critical site in the airway-the surface of the epithelium. We describe a unique strategy to protect the respiratory epithelial cell surface directly by capitalizing on the nondegradative transcytotic pathway of the polymeric immunoglobulin receptor (pIgR). A recombinant fusion protein was constructed consisting of an antihuman pIgR single-chain Fv (scFv) antibody linked to human alpha(1)-antitrypsin (A1AT), an inhibitor of NE. The recombinant scFv-A1AT fusion protein bound specifically to the pIgR on the basolateral surface of an epithelial cell monolayer, and was transported and released into the apical medium where the A1AT domain was capable of forming an inactivation complex with NE. Thus, A1AT linked to an antihuman pIgR scFv was delivered in receptor-specific fashion from the basolateral to apical surface and was released as an active antiprotease, indicating that it is feasible to deliver therapeutic proteins to the apical surface of epithelia by targeting the pIgR.

Author List

Eckman EA, Mallender WD, Szegletes T, Silski CL, Schreiber JR, Davis PB, Ferkol TW

MESH terms used to index this publication - Major topics in bold

Animals
Biological Transport
Cell Line
Dose-Response Relationship, Drug
Epithelial Cells
Humans
Immunoglobulin Fragments
Kinetics
Mice
Receptors, Polymeric Immunoglobulin
Recombinant Fusion Proteins
Transfection
alpha 1-Antitrypsin