Hookworm Infection. Prevalence. In 2002, WHO estimated 1.3 billion infected. 65,000 deaths from its associated anemia. Predominates in tropic and subtropic regions Disease of developing and under-developed world, disease of the poorest of the poor Most vulnerable: Children Pregnant women
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Majority of hookworm infections in humans caused by
1) Ancylostoma duodenale
2) Necator americanus
Soil-transmitted helminthic nematodes
9000 eggs/day, eggs have 3-5 yrs survival
Buccal capsule set with two crescent-shaped cutting plates on ventral side
Ingests 30 µl blood/day
12 mm in length, 0.6 mm in diameter. Female > Male
20,000 eggs/day, eggs have 1 yr survival
Buccal capsule set with symmetric pair of sharp teeth on ventral side.
Ingests 260 µl blood/dayN. americanus and A. duodenale
Find a hookworm antigen that is immunogenic but NOT pathogenic and that can confer protective immunity to infection
Molecular cloning and purification of Ac-TMP, a developmentally regulated putative tissue inhibitor of metalloprotease released in relative abundance by adult Ancylostoma hookworms.
Zhan B. Badamchian M. Meihua B. Ashcom J. Feng J. Hawdon J. Shuhua X. Hotez PJ. American Journal of Tropical Medicine & Hygiene. 66(3):238-44, 2002 Mar.
Used immuno-screening techniques to discover and characterize an adult A. caninum hookworm antigen that could possibly be used in a hookworm vaccine
Grow worms in dog
New phage released
Hookworm Ag released
Incubate with primary antibody
anti-IgG antibodies with marker
Incubate with secondary antibody
anti-secretory protein anti-serum
Identify Secreted Protein
Each peak corresponds to a separated secretory protein
Area under curve = abundance
Secretory proteins separated based on relative hydrophobicity
Fractions containing unique protein
Amino acid sequence
Compare to discovered secreted protein