A hookworm glycoprotein that inhibits neutrophil function is a ligand of the integrin CD11b/CD18.
The chronic survival of many endoparasites is dependent on the ability of these organisms to escape the host immune response. Identification of the molecular mechanisms by which these organisms evade this response may yield novel approaches in the development of anti-inflammatory agents. We describe here the discovery and characterization of a novel 41-kilodalton glycoprotein from the canine hookwork (Ancylostoma caninum) that potently inhibits CD11/CD18-dependent neutrophil function in vitro. Neutrophil inhibitory factor (NIF) blocks the adhesion of activated human neutrophils to vascular endothelial cells as well as the release of H2O2 from activated neutrophils, over a similar concentration range (IC50 10-20 nM). Studies aimed at determining the nature of the NIF binding site on neutrophils revealed selective, high affinity binding of this protein to the integrin CD11b/CD18. A cDNA encoding NIF was isolated from a canine hookworm cDNA library. NIF comprises a mature polypeptide of 257 amino acids, preceded by a 17-amino acid leader. The mature protein has 10 cysteines and has seven potential N-linked glycosylation sites. NIF has no significant sequence homologies to any previously reported protein. As such, NIF represents a prototype of a novel class of leukocyte function inhibitors.
Moyle
M
M
Corvas International Inc., San Diego, California 92121.
Foster
D L
DL
McGrath
D E
DE
Brown
S M
SM
Laroche
Y
Y
De Meutter
J
J
Stanssens
P
P
Bogowitz
C A
CA
Fried
V A
VA
Ely
J A
JA
D000595:Amino Acid Sequence; D000723:Ancylostomatoidea / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D001483:Base Sequence; D018821:CD18 Antigens; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D004285:Dogs; D006023:Glycoproteins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D015801:Helminth Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007962:Leukocytes / Q000187:drug effects / Q000502:physiology; D016177:Macrophage-1 Antigen / Q000378:metabolism*; D008565:Membrane Proteins / Q000097:blood; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology
D015703:Antigens, CD; D018821:CD18 Antigens; D017931:DNA Primers; D018076:DNA, Complementary; D006023:Glycoproteins; D015801:Helminth Proteins; D016177:Macrophage-1 Antigen; D008565:Membrane Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C086581:NIF protein, Ancylostoma caninum