Anti-receptor antibodies inhibit Pseudomonas aeruginosa binding to the cornea and prevent corneal perforation.
A polyclonal antibody (pAb) against gangliotetraosylceramide (asialo GM1), a glycolipid to which bacterial pili and LPS bind, and a mAb against a 66 kDa pilus-binding protein purified from adult mouse corneal epithelium were used to determine if antibodies against host receptors for bacterial adhesins could inhibit bacterial binding to wounded corneal epithelium and protect ocularly challenged mice from corneal perforation when topically applied. Bacteria were mixed with anti-66 kDa mAb, a mixture of anti-asialo GM1 pAb and anti-66 kDa mAb, an irrelevant control mAb (anti-human histocompatibility Ag HLA-DR5) or PBS prior to application to scarified corneas in organ culture. The combination of the two antibodies or the anti-66 kDa mAb alone was effective in reducing bacterial adherence compared with either PBS or the antibody control. To determine if these antibodies were protective in vivo, corneas of C57BL/6J mice were scarified and inoculated with Pseudomonas aeruginosa. Eyes were treated topically with anti-asialo GM1 pAb, anti-66 kDa mAb, a mixture of the two or control mouse serum. More serum-treated corneas perforated compared to corneas from any other group (P < or = 0.005) by 30 days postinfection. Treatment with a combination of the two antibodies resulted in significantly less corneal pathology 30 days p.i. when compared to any other treatment (P < or = 0.005). These data provide evidence that antibodies against host corneal receptors significantly inhibit bacterial binding in vitro and when applied topically in vivo, lessen the severity of ocular disease characteristic of P. aeruginosa keratitis.
Hobden
J A
JA
Department of Anatomy and Cell Biology, Wayne State University, School of Medicine, Detroit, Michigan 48201, USA.
Gupta
S K
SK
Masinick
S A
SA
Wu
X
X
Kernacki
K A
KA
Berk
R S
RS
Hazlett
L D
LD
D000818:Animals; D000907:Antibodies, Bacterial / Q000494:pharmacology*; D001667:Binding, Competitive / Q000276:immunology; D015180:Electrophoresis, Gel, Two-Dimensional; D004848:Epithelium / Q000276:immunology; D015807:Eye Injuries, Penetrating / Q000276:immunology* / Q000517:prevention & control*; D005260:Female; D010861:Fimbriae, Bacterial / Q000737:chemistry; D005677:G(M1) Ganglioside / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D011550:Pseudomonas aeruginosa / Q000276:immunology* / Q000502:physiology*; D011971:Receptors, Immunologic / Q000276:immunology*
D000907:Antibodies, Bacterial; D011971:Receptors, Immunologic; D005677:G(M1) Ganglioside; C018835:asialo GM1 ganglioside