title: Transcriptome profiling of gene expression during immunisation trial against Fasciola hepatica: identification of genes and pathways involved in conferring immunoprotection in a murine model.
abstract: Fasciolosis remains a significant food-borne trematode disease causing high morbidity around the world and affecting grazing animals and humans. A deeper understanding concerning the molecular mechanisms by which Fasciola hepatica infection occurs, as well as the molecular basis involved in acquiring protection is extremely important when designing and selecting new vaccine candidates. The present study provides a first report of microarray-based technology for describing changes in the splenic gene expression profile for mice immunised with a highly effective, protection-inducing, multi-epitope, subunit-based, chemically-synthesised vaccine candidate against F. hepatica.
  
  
  
  The mice were immunised with synthetic peptides containing B- and T-cell epitopes, which are derived from F. hepatica cathepsin B and amoebapore proteins, as novel vaccine candidates against F. hepatica formulated in an adjuvant adaptation vaccination system; they were experimentally challenged with F. hepatica metacercariae. Spleen RNA from mice immunised with the highest protection-inducing synthetic peptides was isolated, amplified and labelled using Affymetrix standardised protocols. Data was then background corrected, normalised and the expression signal was calculated. The Ingenuity Pathway Analysis tool was then used for analysing differentially expressed gene identifiers for annotating bio-functions and constructing and visualising molecular interaction networks.
  
  
  
  Mice immunised with a combination of three peptides containing T-cell epitopes induced high protection against experimental challenge according to survival rates and hepatic damage scores. It also induced differential expression of 820 genes, 168 genes being up-regulated and 652 genes being down-regulated, p value <0.05, fold change ranging from -2.944 to 7.632. A functional study of these genes revealed changes in the pathways related to nitric oxide and reactive oxygen species production, Interleukin-12 signalling and production in macrophages and Interleukin-8 signalling with up-regulation of S100 calcium-binding protein A8, Matrix metallopeptidase 9 and CXC chemokine receptor 2 genes.
  
  
  
  The data obtained in the present study provided us with a more comprehensive overview concerning the possible molecular pathways implied in inducing protection against F. hepatica in a murine model, which could be useful for evaluating future vaccine candidates.
authors.affiliation: Parasite and Molecular Immunology Laboratory, Tropical Disease Research Centre, (IBSAL-CIETUS), University of Salamanca, Salamanca, Spain.|Molecular Biology and Immunology Department, Fundación Instituto de Inmunología de Colombia (FIDIC), Bogotá, Colombia.|Present address: Centro de Investigación en Salud para el Trópico (CIST), Facultad de Medicina, Universidad Cooperativa de Colombia, Santa Marta, Magdalena, Colombia.
authors.forename: Jose
authors.initials: J
authors.lastname: Rojas-Caraballo
authors.affiliation: Parasite and Molecular Immunology Laboratory, Tropical Disease Research Centre, (IBSAL-CIETUS), University of Salamanca, Salamanca, Spain.
authors.forename: Julio
authors.initials: J
authors.lastname: López-Abán
authors.affiliation: Molecular Biology and Immunology Department, Fundación Instituto de Inmunología de Colombia (FIDIC), Bogotá, Colombia.|Basic Sciences Department, School of Medicine and Health Sciences, Universidad del Rosario, Bogotá, Colombia.
authors.forename: Darwin Andrés
authors.initials: DA
authors.lastname: Moreno-Pérez
authors.affiliation: Parasite and Molecular Immunology Laboratory, Tropical Disease Research Centre, (IBSAL-CIETUS), University of Salamanca, Salamanca, Spain.
authors.forename: Belén
authors.initials: B
authors.lastname: Vicente
authors.affiliation: Parasite and Molecular Immunology Laboratory, Tropical Disease Research Centre, (IBSAL-CIETUS), University of Salamanca, Salamanca, Spain.
authors.forename: Pedro
authors.initials: P
authors.lastname: Fernández-Soto
authors.affiliation: Pharmaceutical Chemistry Department, (IBSAL-CIETUS), University of Salamanca, Salamanca, Spain.
authors.forename: Esther
authors.initials: E
authors.lastname: Del Olmo
authors.affiliation: Molecular Biology and Immunology Department, Fundación Instituto de Inmunología de Colombia (FIDIC), Bogotá, Colombia.|Basic Sciences Department, School of Medicine and Health Sciences, Universidad del Rosario, Bogotá, Colombia.
authors.forename: Manuel Alfonso
authors.initials: MA
authors.lastname: Patarroyo
authors.affiliation: Parasite and Molecular Immunology Laboratory, Tropical Disease Research Centre, (IBSAL-CIETUS), University of Salamanca, Salamanca, Spain. ama@usal.es.
authors.forename: Antonio
authors.initials: A
authors.lastname: Muro
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology; D040501:Calgranulin A / Q000187:drug effects / Q000235:genetics; D000939:Epitopes / Q000276:immunology; D005210:Fasciola hepatica / Q000276:immunology*; D005211:Fascioliasis / Q000517:prevention & control*; D005260:Female; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D018664:Interleukin-12 / Q000235:genetics; D016209:Interleukin-8 / Q000187:drug effects / Q000235:genetics; D020780:Matrix Metalloproteinase 9 / Q000187:drug effects / Q000235:genetics; D051379:Mice; D010455:Peptides / Q000276:immunology; D016052:Protozoan Vaccines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D023063:Receptors, Interleukin-8B / Q000187:drug effects / Q000235:genetics; D013154:Spleen / Q000187:drug effects* / Q000378:metabolism; D015854:Up-Regulation; D014611:Vaccination
chemical_list: D000909:Antibodies, Helminth; C508606:CXCL8 protein, human; D040501:Calgranulin A; D000939:Epitopes; D016209:Interleukin-8; D010455:Peptides; D016052:Protozoan Vaccines; D012333:RNA, Messenger; D023063:Receptors, Interleukin-8B; C086145:S100a8 protein, mouse; D018664:Interleukin-12; D020780:Matrix Metalloproteinase 9; C501830:Mmp9 protein, mouse
keywords: Epitope; Fasciolosis; Gene expression; Immunology; Microarrays; Vaccine