title: Production of a bovine IL-12p40 probe and application using in situ hybridization on ruminant fixed tissues.
abstract: Pro-inflammatory cytokines (particularly IL-12) are important for initiating protective T helper 1 (Th1)-type immune responses and hence vital for combating intracellular infections and tumours. In situ hybridization (ISH) provides a powerful diagnostic tool allowing the identification and localization of cells producing these mediators in fixed tissues. The objective of this work was to produce a bovine IL-12p40 probe that allows detection of IL-12p40 mRNA in fixed tissues from different ruminant species. The RNA probe sequence is 447bp in length and from a region with high cross-species-sequence homology (>97.3% homology) to the ovine, cervine, caprine and bubaline IL-12p40 genes. ISH was carried out on paraformaldehyde fixed tissues collected from cattle, sheep and goats. The probe was efficient in identifying IL-12p40 expressing cells in fixed tissues from all these species. In conclusion, the IL-12p40 probe was efficient in identifying and localizing cells that express IL-12p40, and provides a good immuno-diagnostic technique to characterize immune responses in fixed tissues.
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Bush Loan, EH26 0PZ Edinburgh, United Kingdom. german.canton@moredun.ac.uk
authors.forename: Germán
authors.initials: G
authors.lastname: Cantón
authors.forename: Paul
authors.initials: P
authors.lastname: Bartley
authors.forename: Kathryn
authors.initials: K
authors.lastname: Bartley
authors.forename: Helen
authors.initials: H
authors.lastname: Todd
authors.forename: Francesca
authors.initials: F
authors.lastname: Chianini
authors.forename: Frank
authors.initials: F
authors.lastname: Katzer
mesh_terms: D000818:Animals; D001483:Base Sequence; D002020:Buffaloes / Q000235:genetics / Q000276:immunology; D002417:Cattle; D003670:Deer / Q000235:genetics / Q000276:immunology; D005260:Female; D006041:Goats / Q000235:genetics / Q000276:immunology; D017403:In Situ Hybridization / Q000662:veterinary; D053762:Interleukin-12 Subunit p40 / Q000235:genetics*; D015336:Molecular Probe Techniques / Q000662:veterinary*; D011247:Pregnancy; D015347:RNA Probes / Q000096:biosynthesis* / Q000235:genetics*; D012418:Ruminants / Q000235:genetics* / Q000276:immunology*; D012756:Sheep / Q000235:genetics / Q000276:immunology; D013045:Species Specificity; D018417:Th1 Cells / Q000276:immunology
chemical_list: D053762:Interleukin-12 Subunit p40; D015347:RNA Probes