Possible Role for Toll-Like Receptors in Interaction of Fasciola hepatica Excretory-Secretory Products with Human Monocyte Cell Line.

ABSTRACT

This chapter presents a proteomic approach to purify and identify native excretory-secretory products (ESPs) in the range of >10-30 kDa proteins capable of interacting with toll-like receptors (TLRs). Here we present a protocol to fractionate the total ESPs using an ultrafiltration system to recover ESP proteins >10-30 kDa. The fraction of the proteins >10-30 kDa is purified by ion exchange chromatography (IEC) using a mono Q-column in a fast protein liquid chromatography system (FPLC) to separate its components based on charge. Finally, a screening system is presented using THP1-Blue CD14 cells to investigate whether TLRs could also be targeted by Fasciola hepatica ESPs and the interaction with TLR4 using HEK293 Blue-TLR4 cells.