Mass spectrometry analysis of the excretory-secretory (E-S) products of the model cestode Hymenolepis diminuta reveals their immunogenic properties and the presence of new E-S proteins in cestodes

Hymenolepis diminuta is an important model species in studies of therapeutics, biochemical processes, immune responses and other aspects of cestodiasis. The parasite produces numerous excretory-secretory (E-S) proteins and a glycocalyx covering its body. Our study focused on the mass spectrometry analysis of the E-S material with an objective to determine if E-S contains any new proteins, in particular those that can be identified as: antigens, vaccine candidates and drug targets. These proteins might engage directly in host-parasite interactions. Adult parasites collected from experimentally infected rats were cultured in vitro for 5 and 18h. Immunoblotting was used to verify which E-S protein bands separated in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) react with specific antibodies from sera of infected rats. We identified thirty-nine proteins by LC-MS/MS (liquid chromatography mass spectrometry). Results indicated the presence of proteins that have never been identified in cestode E-S material. Immunoblotting showed the immunogenicity of E-S products of H. diminuta, most probably associated with the presence of proteins known as antigens in other flatworm species. Among identified proteins are those engaged in immunomodulatory processes (eg. HSP), in response to oxidative stress (peroxidasin) or metabolism (eg. GAPDH). The predominant functions are associated with metabolism and catalytic activity. This is the first study identifying E-S-proteins in adult tapeworms, thus providing information for better understanding host-parasite interrelationships, and may point out potential targets for vaccines or drug discovery studies, as among the proteins observed in our study are those known to be antigens.