title: Influence of subculturing on gene expression in a Theileria lestoquardi-infected cell line.
abstract: In this study potential molecular markers for identification of attenuation in a Theileria lestoquardi-infected cell line to be used in vaccination trials were identified. Two markers associated with attenuation in Theileria annulata vaccine strains were analyzed (metalloproteinase activity and TNF? mRNA expression). The result showed a decreased activity of MMP 9 and decreased mRNA expression of TNF? with increasing passage number. Suppression subtractive hybridization was used to identify potential new markers of attenuation. Random screening revealed nine differentially expressed genes, one from the parasite and eight from the host. Quantitative real time-PCR confirmed mRNA expression of the parasite vacuolar H+ATPase to be downregulated at higher passages.
authors.affiliation: Division of Veterinary Infection Biology and Immunology, Research Center Borstel, Parkallee 22, 23845 Borstel, Germany.
authors.forename: Awadia M
authors.initials: AM
authors.lastname: Ali
authors.forename: Doreen
authors.initials: D
authors.lastname: Beyer
authors.forename: Mohammed A
authors.initials: MA
authors.lastname: Bakheit
authors.forename: Birgit
authors.initials: B
authors.lastname: Kullmann
authors.forename: Diaeldin A
authors.initials: DA
authors.lastname: Salih
authors.forename: Jabbar S
authors.initials: JS
authors.lastname: Ahmed
authors.forename: Ulrike
authors.initials: U
authors.lastname: Seitzer
mesh_terms: D000818:Animals; D002417:Cattle; D002460:Cell Line; D005786:Gene Expression Regulation; D054884:Host-Pathogen Interactions; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D012692:Serial Passage; D012756:Sheep; D016794:Theileria / Q000254:growth & development* / Q000472:pathogenicity*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; D014409:Tumor Necrosis Factor-alpha; D020780:Matrix Metalloproteinase 9; D025262:Vacuolar Proton-Translocating ATPases