Isolation and molecular cloning of a secreted hookworm platelet inhibitor from adult Ancylostoma caninum.
Hookworms, bloodfeeding intestinal nematodes, are a leading cause of iron deficiency anemia in the developing world. These parasites have evolved potent mechanisms of interfering with mammalian hemostasis, presumably for the purpose of facilitating bloodfeeding. Adult Ancylostoma caninum worm extracts contain an activity that inhibits platelet aggregation and adhesion by blocking the function of two cell surface integrin receptors, Glycoprotein IIb/IIIa and GPIa/IIa. Using rpHPLC, the hookworm platelet inhibitor activities have been purified from protein extracts of A. caninum. Because the two inhibitory activities co-purified through multiple chromatographic steps, have similar molecular masses and share identical N-terminal as well as internal amino acid sequence homology, it is likely that they represent a single gene product. A cDNA corresponding to the purified hookworm platelet inhibitor (HPI) protein has been cloned from adult A. caninum RNA, and the translated amino acid sequence shows significant homology to Neutrophil Inhibitory Factor and Ancylostoma Secreted Proteins, suggesting that these related hookworm proteins represent a novel class of integrin receptor antagonists. Polyclonal antibodies raised against the recombinant HPI protein recognize corresponding native proteins in A. caninum extracts and excretory/secretory products, and immunohistochemistry data have identified the cephalic glands as the major source of the inhibitor within the adult hookworm. These data suggest that HPI is secreted by the adult stage of the parasite at the site of intestinal attachment. As such, it may represent a viable target for a vaccine-based strategy aimed at interfering with hookworm-induced gastrointestinal hemorrhage and iron deficiency anemia.
Del Valle
Antonio
A
Department of Pediatrics, Yale University School of Medicine, 464 Congress Avenue, New Haven, CT 06520-8081, USA.
Jones
Brian F
BF
Harrison
Lisa M
LM
Chadderdon
Robert C
RC
Cappello
Michael
M
D000595:Amino Acid Sequence; D000722:Ancylostoma / Q000737:chemistry* / Q000235:genetics; D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D003094:Collagen / Q000378:metabolism; D018076:DNA, Complementary; D005340:Fibrinogen / Q000378:metabolism; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000494:pharmacology; D007074:Immunoglobulin G / Q000276:immunology; D007150:Immunohistochemistry; D038982:Integrin alpha2beta1 / Q000378:metabolism; D008969:Molecular Sequence Data; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D021241:Spectrometry, Mass, Electrospray Ionization
D000909:Antibodies, Helminth; D018076:DNA, Complementary; C476807:HPI-1 protein, Ancylostoma caninum; D015801:Helminth Proteins; D007074:Immunoglobulin G; D038982:Integrin alpha2beta1; D010975:Platelet Aggregation Inhibitors; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D011994:Recombinant Proteins; D005340:Fibrinogen; D003094:Collagen