Isolation and Characterization of Angiostrongylus cantonensis L5 Excretory-Secretory Proteins

Live L5 of A. cantonensis were isolated from the brain tissues of rats by anesthetizing with 30 μl Zoletil 50 (Virbac) after 21 days post infection. After the worms were washed with saline, phosphate-buffered saline, distilled water and RPMI (Sigma-Aldrich, St. Louis, USA), they were incubated in RPMI without foetal bovine serum (FBS) for 24, 48 and 72 h. The ESP of L5 were collected and concentrated by Amicon Ultra-15 10 K centrifugal filter devices (Merck Millipore, Darmstadt, Germany) from the culture medium. The ESP concentration in the medium was determined using a Bio-Rad Protein Assay Kit (Bio-Rad, Hercules, CA, USA), according to the manufacturer’s instructions. The ESP were used to treat the astrocytes, and cellular changes were observed [34 (link)].