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Targeting mucosal sites by polymeric immunoglobulin receptor-directed peptides.

White KD, Capra JD - J. Exp. Med. (2002)

Bottom Line: Here by binding human secretory component to overlapping decapeptides of Calpha3, we confirm these residues and also uncover an additional site.Some transcytosis-selected peptides map to the same 402-410 pIgR-binding Calpha3 site.Further in vivo studies document that at least one of these peptides is transported in a rat model measuring hepatic bile transport.

View Article: PubMed Central - PubMed

Affiliation: Molecular Immunogenetics Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK 73104, USA.

ABSTRACT
Polymeric immunoglobulins provide first line humoral defense at mucosal surfaces to which they are specifically transported by the polymeric immunoglobulin receptor (pIgR) on mucosal and glandular epithelial cells. Previous studies from our laboratory suggested that amino acids 402-410 of the Calpha3 domain of dimeric IgA (dIgA) represented a potential binding site for the pIgR. Here by binding human secretory component to overlapping decapeptides of Calpha3, we confirm these residues and also uncover an additional site. Furthermore, we show that the Calpha3 motif appears to be sufficient to direct transport of green fluorescent protein through the pIgR-specific cellular transcytosis system. An alternative approach identified phage peptides, selected from a library by the in vitro Madin Darby Canine Kidney transcytosis assay, for pIgR-mediated transport through epithelial cells. Some transcytosis-selected peptides map to the same 402-410 pIgR-binding Calpha3 site. Further in vivo studies document that at least one of these peptides is transported in a rat model measuring hepatic bile transport. In addition to identifying small peptides that are both bound and transported by the pIgR, this study provides evidence that the pIgR-mediated mucosal secretion system may represent a means of targeting small molecule therapeutics and genes to mucosal epithelial cells.

Show MeSH
Kinetics of IPS phage peptide secretion into rat bile. The bile ducts of male Wistar rats were cannulated. Bile collection began after I.V. injection of 106 specific phage (IPS) and 106 negative control phage particles and was continued for 30 min time points in separate tubes. Phage titers from each time point were determined by standard procedures.
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fig6: Kinetics of IPS phage peptide secretion into rat bile. The bile ducts of male Wistar rats were cannulated. Bile collection began after I.V. injection of 106 specific phage (IPS) and 106 negative control phage particles and was continued for 30 min time points in separate tubes. Phage titers from each time point were determined by standard procedures.

Mentions: To determine if the phage-displayed peptides are transported in vivo, a mixture of pIgR transcytosis-selected IPS phage and nonspecific phage were injected intravenously into rats. Bile was collected at 30 min time intervals to measure hepatic transport. The kinetics of IPS phage transport were similar to that reported previously for dIgA (16; Fig. 6) . 100% of the phage recovered from all time points displayed the IPS peptide (unpublished data). No nonspecific phage were recovered at any time point (unpublished data). Therefore, at least one of the phage peptides identified by in vitro transcytosis is transported specifically by the pIgR in vivo in a rat model for pIgR transport.


Targeting mucosal sites by polymeric immunoglobulin receptor-directed peptides.

White KD, Capra JD - J. Exp. Med. (2002)

Kinetics of IPS phage peptide secretion into rat bile. The bile ducts of male Wistar rats were cannulated. Bile collection began after I.V. injection of 106 specific phage (IPS) and 106 negative control phage particles and was continued for 30 min time points in separate tubes. Phage titers from each time point were determined by standard procedures.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196054&req=5

fig6: Kinetics of IPS phage peptide secretion into rat bile. The bile ducts of male Wistar rats were cannulated. Bile collection began after I.V. injection of 106 specific phage (IPS) and 106 negative control phage particles and was continued for 30 min time points in separate tubes. Phage titers from each time point were determined by standard procedures.
Mentions: To determine if the phage-displayed peptides are transported in vivo, a mixture of pIgR transcytosis-selected IPS phage and nonspecific phage were injected intravenously into rats. Bile was collected at 30 min time intervals to measure hepatic transport. The kinetics of IPS phage transport were similar to that reported previously for dIgA (16; Fig. 6) . 100% of the phage recovered from all time points displayed the IPS peptide (unpublished data). No nonspecific phage were recovered at any time point (unpublished data). Therefore, at least one of the phage peptides identified by in vitro transcytosis is transported specifically by the pIgR in vivo in a rat model for pIgR transport.

Bottom Line: Here by binding human secretory component to overlapping decapeptides of Calpha3, we confirm these residues and also uncover an additional site.Some transcytosis-selected peptides map to the same 402-410 pIgR-binding Calpha3 site.Further in vivo studies document that at least one of these peptides is transported in a rat model measuring hepatic bile transport.

View Article: PubMed Central - PubMed

Affiliation: Molecular Immunogenetics Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK 73104, USA.

ABSTRACT
Polymeric immunoglobulins provide first line humoral defense at mucosal surfaces to which they are specifically transported by the polymeric immunoglobulin receptor (pIgR) on mucosal and glandular epithelial cells. Previous studies from our laboratory suggested that amino acids 402-410 of the Calpha3 domain of dimeric IgA (dIgA) represented a potential binding site for the pIgR. Here by binding human secretory component to overlapping decapeptides of Calpha3, we confirm these residues and also uncover an additional site. Furthermore, we show that the Calpha3 motif appears to be sufficient to direct transport of green fluorescent protein through the pIgR-specific cellular transcytosis system. An alternative approach identified phage peptides, selected from a library by the in vitro Madin Darby Canine Kidney transcytosis assay, for pIgR-mediated transport through epithelial cells. Some transcytosis-selected peptides map to the same 402-410 pIgR-binding Calpha3 site. Further in vivo studies document that at least one of these peptides is transported in a rat model measuring hepatic bile transport. In addition to identifying small peptides that are both bound and transported by the pIgR, this study provides evidence that the pIgR-mediated mucosal secretion system may represent a means of targeting small molecule therapeutics and genes to mucosal epithelial cells.

Show MeSH