Influence of subculturing on gene expression in a Theileria lestoquardi-infected cell line

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 alpha mRNA expression). The result showed a decreased activity of MMP 9 and decreased mRNA expression of TNF alpha with increasing passage number. Suppression subtractive hybridization was used to identify potential new markets 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. (c) 2008 Elsevier Ltd. All rights reserved.