EOTAXIN IS SPECIFICALLY CLEAVED BY HOOKWORM METALLOPROTEASES PREVENTTING ITS ACTION IN VITRO AND IN VIVO
Brooke Pottorff
Parasite survival in the host
Introduction: immunity of parasites
OBJECTIVE:
To show that hookworms produce a kind of protease (specifically a metalloprotease) that inactivates eotaxin
BACKGROUND
Eotaxin is a chemokine that attracts eosinophils to the site of infection
Inactivation of eotaxin enables the hookworm to avoid the innate immune system
objectives
Materials and Methods
Objective 1- Production of NES
Microscopic N. americanus hookworm was injected into the skin of hamsters
Retrieved worms and incubated them on RPMI media
Liquid media is similar to their living conditions in mammalian cell
Worms were able to grow and produce excretory/secretory products (NES)
BACKGROUND:
- Life of a hookworm
Adult hookworm attached to intestinal wall
Objective 2- response of eosinophils to NES in vivo
Eosinophils taken from guinea pigs and radiolabeled
Then injected into recipient guinea pigs after being mixed with various combinations of substances (x-axis)
measured eosinophil accumulation at sites
(since eotaxin is a chemoattractant there should be lots of eosinophils in the presence of eotaxin)
Using ELISA to measure amount of Eotaxin
Western blot to determine the specificity of NES for different chemokines (objective 3)
N. Americanus homogenate
Importance of this study
Hookworm thought to infect 800 million people world wide
Cause anemia and protein loss as they suck blood from the gut
Can cause intellectual, cognitive and growth defects in children
Treatment can target the proteases secreted by parasites