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RGDSK Peptide Functionalized Helical Rosette Nanotubes (RGDSK‐HRNs) Block Integrin αvβ3 and Inhibit E. coli Adherence to Intestinal Porcine Epithelial 1 Cell Line (IPEC1) in vitro
Author(s) -
Le Nguyen Phuong Khanh,
Quach Chi Cuong,
Aulakh Gurpreet,
Gerdts Volker,
Fenniri Hicham,
Singh Baljit
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.658.10
Subject(s) - integrin , microbiology and biotechnology , adhesion , integrin alpha m , biology , cell adhesion , epithelium , intestinal epithelium , chemistry , cell , biochemistry , flow cytometry , genetics , organic chemistry
E. coli infection is a leading cause of intestinal disease and death, and inflicts billions of dollars in economic losses. There is ongoing effort to find ways to reduce intestinal colonization by E. coli . Integrin αvβ3, recognizing arginine‐glycine‐aspartic acid (RGD) sequences, has important functions in cell adhesion, signaling and survival. However, the expression and role of integrin αvβ3 in the adhesion of E. coli on intestinal epithelium remain unknown. We used porcine jejunum, IPEC1 cells and E. coli K88 to firstly explored the hitherto unknown expression of integrin αvβ3 with light, fluorescent and electron immunocytochemistry, immunoprecipitation and Western blots. We then used RGDSK peptide and RGDSK‐HRNs to test the role of integrin αvβ3 in adhesion of E. coli K88 to IPEC1 cells. Normal porcine jejunum strongly expresses integrin αvβ3 on the nucleus and apical surface of epithelium and gland cells. The expression of integrin αvβ3 decreased in the epithelium but increased on vascular endothelium of the jejunum infected with E. coli or E. coli associated with Salmonella . Integrin αvβ3 was found on plasma membrane, cytoplasm, and nucleus of IPEC1 cells. The expression of integrin αvβ3 on IPEC1 decreased at 15 minutes but returned to normal after 90 minutes of infection with E. coli K88 (P<0.05). Immuno‐gold quantification showed changes in sub‐cellular expression of the integrin αvβ3 in IPEC1. The light, fluorescent and electron microscopy, and western blots showed novel expression of integrin αvβ3 on E. coli K88. Finally, we found that RGDSK peptides and RGDSK‐HRNs bind integrin αvβ3 on both IPEC1 and E. coli K88. RGDSK peptides block the attachment of E. coli to IPEC1 (5 minutes IPEC1 exposed to 50μM RGDSK, P<0.05), and reduce survival of E. coli (10 minutes IPEC1 exposed to 50μM RGDSK, P<0.05). Dose dependent RGDSK‐HRNs mediates the attachment of E. coli to IPEC1 more efficiently and regularly (10 minutes IPEC1 exposed to HRNs containing equivalent 5μM RGDSK, P<0.001). These are the first data to show a role for the integrin αvβ3 in the attachment of E. coli to the intestinal epithelium, and that novel RGDSK helical rosette nanotubes can inhibit the attachment of E. coli to the epithelium. Support or Funding Information Saskatchewan Agriculture Development Fund and NSERC