Project description:We have employed whole genome microarray expression profiling to identify differently expressed genes following infection of interleukin-16-knock-out bone-marrow derived macrophages with T. whipplei. Macrophages were infected with T. whipplei (MOI 50:1) for 6 hours and a signature was identified that distinguished between infected and control samples. Expression of several genes from this signature was quantified in the same RNA samples by real-time PCR, confirming the predicted macrophage response pattern.
Project description:We have employed whole genome microarray expression profiling to identify differently expressed genes following infection of interleukin-16-knock-out bone-marrow derived macrophages with T. whipplei. Macrophages were infected with T. whipplei (MOI 50:1) for 6 hours and a signature was identified that distinguished between infected and control samples. Expression of several genes from this signature was quantified in the same RNA samples by real-time PCR, confirming the predicted macrophage response pattern. Gene expression in macrophages was measured at 6 hours after stimulation Three independent experiments were performed.
Project description:Whipple's disease (WD) affects only a very small minority of individuals infected by Tropheryma whipplei (Tw). Asymptomatic and chronic carriage of the causative organism is less rare and therefore, the pathogenesis of WD is poorly understood. Here we studied transcriptome responses in peripheral blood mononuclear cells (PBMCs) that were obtained from members of a large multiplex French kindred including otherwise healthy WD patients, healthy chronic carriers of Tw and other unrealted control subjects.
Project description:We have employed whole genome microarray expression profiling to identify differently expressed genes following stimulation of interleukin-16-knock-out bone-marrow derived macrophages with Escherichia coli lipopolysaccharide (LPS). Macrophages were stimulated with LPS (100 ng/ml) for 6 hours and a signature was identified that distinguished between infected and control samples. Expression of several genes from this signature was quantified in the same RNA samples by real-time PCR, confirming the predicted macrophage response pattern.
Project description:We have employed whole genome microarray expression profiling to identify differently expressed genes following stimulation of interleukin-16-knock-out bone-marrow derived macrophages with Escherichia coli lipopolysaccharide (LPS). Macrophages were stimulated with LPS (100 ng/ml) for 6 hours and a signature was identified that distinguished between infected and control samples. Expression of several genes from this signature was quantified in the same RNA samples by real-time PCR, confirming the predicted macrophage response pattern. Gene expression in macrophages was measured at 6 hours after stimulation Three independent experiments were performed.
Project description:In this work we analyzed the genomic diversity of several Tropheryma whipplei strains by microarray based-comparative genomic hybridization. Fifteen clinical isolates originating from biopsy samples recovered from different countries were compared with the T. whipplei Twist strain. For each isolate, the genes were defined as either present or absent/divergent using the GACK analysis software. Genomic changes were then further characterized by PCR and sequencing. Obtained results revealed a limited genetic variation between these T. whipplei isolates with at most 2.24 % of the probes exhibiting differential hybridization against the Twist strain. The main variation was found in genes encoding for the WiSP family proteins supporting the view of these membrane proteins as key actors of immune evasion. This work also evidenced a 19.2 kb-pair deletion within T. whipplei DIG15 strain. This deletion takes place in the same region as the large genomic rearrangement previously described between Twist and TW08/27 which can thus be considered as a major hot-spot for intra-specific T. whipplei differentiation. Analysis of this deleted region confirmed the role of WND-domains in generating T. whipplei diversity Keywords: Comparative genomic hybridization
