ABSTRACT
Changes and local immune response were evaluated in the peritoneal cell populations, duodenal lamina propria and liver from goats immunized with recombinant glutathione transferase sigma class (rFhGST-S1) during early stages of infection with Fasciola hepatica. Group 1 (n=7) was unimmunized and uninfected; group 2 (n=10) was immunized with adjuvant Quil A and infected; group 3 (n=10) was immunised with rFhGST-S1 and infected. Three goats from each group were killed at 7-9 days post-infection (dpi) to evaluate early changes and immune response. The remaining goats were killed at 15 weeks post-infection (wpi). rFhGST-S1 vaccination induced variable response: three goats showed low fluke burden at 15 wpi and two goats showed low hepatic damage at early infection stages. This response was associated to a severe infiltrate of eosinophils in peritoneal fluid and hepatic necrotic foci, high iNOS expression in peritoneal cells and abundant infiltrate of eosinophils surrounding hepatic migrating flukes. T lymphocyte subsets were found in the vicinity of necrotic areas but they were absent in the vicinity of migrating larvae. No significant variation for T cell subsets, except for CD4 and γδ T lymphocytes, that were higher in the Quil A group compared to the rFhGST-S1 group. Expression of IL4 and IFN-γ in the hepatic inflammatory infiltrates was very occasional.
Subject(s)
Fasciola hepatica/immunology , Fascioliasis/veterinary , Goat Diseases/parasitology , Liver/pathology , Peritoneum/pathology , Vaccines/immunology , Adjuvants, Immunologic/therapeutic use , Animals , Fascioliasis/immunology , Fascioliasis/parasitology , Fascioliasis/pathology , Fascioliasis/prevention & control , Glutathione Transferase/genetics , Glutathione Transferase/immunology , Goat Diseases/immunology , Goat Diseases/pathology , Goat Diseases/prevention & control , Goats/immunology , Goats/parasitology , Immunity, Cellular/immunology , Immunity, Humoral/immunology , Liver/parasitology , Male , Peritoneum/parasitology , Quillaja Saponins , Saponins/therapeutic use , Vaccines/therapeutic use , Vaccines, Synthetic/immunology , Vaccines, Synthetic/therapeutic useABSTRACT
Loss-of-function phenotypic analysis via interference RNA (RNAi) technology is a revolutionary approach to assigning gene function. While transcript-based methodologies commonly validate RNAi gene suppression investigations, protein-based validation is less developed. This report illustrates the potential for two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2-DE) and gel analysis to quantify protein levels following RNAi. This case study involves three glutathione transferase (GST) genes targeted by RNAi from the model organism Caenorhabditis elegans.
