MicroRNA profiling of human cholangiocyte H69 cells after infection with Cryptosporidium parvum
ABSTRACT: H69 cells were cultured in H69 medium with Cryptosporidium parvum oocysts(10 X 5 per well, for smaples 04, 05 and 06) or without oocysts(for samples 01, 02 and 03)for 8 hours and then collected for array analysis. Sample 07 was cells exposed to heated inactived oocysts.
Cryptosporidium parvum is a protozoan parasite that infects the gastrointestinal epithelium and causes diarrheal disease worldwide. Innate epithelial immune responses are key mediators of the host's defense to C. parvum. MicroRNAs (miRNAs) regulate gene expression at the posttranscriptional level and are involved in regulation of both innate and adaptive immune responses. Using an in vitro model of human cryptosporidiosis, we analyzed C. parvum-induced miRNA expression in biliary epithelial cell ...[more]
Project description:H69 cells were cultured in H69 medium with 1 ng/ml lipopolysaccharide(LPS, for smaples 04, 05 and 06) or without LPS(for samples 01, 02 and 03) for 8 hours and then collected for array analysis. <br>
Project description:Cells of a human cholangiocyte cell line (H69) were exposed to 10 ng/ml of IFN-gamma for 8 h. Cells were collected and processed for miRNA array analysis carried out by EXIQON (Denmark).
Project description:Global gene expression in C. parvum environmental stage (oocysts) and the oocysts treated with UV comparing control untreated ones. Goal was to uncover the metabolic features in oocysts and the oocysts treated with UV. two-condition experiment, UV treatment vs. UV untreatment; two time points, 0.5h and 5h. Each time point, two Biological replicates(1, 2) with two technique replicates(1-1,1-2 ; 2-1, 2-2).
Project description:To date little is known about the transcriptome of Hammondia hammondi, the nearest extant relative of Toxoplasma gondii. In this study we used an existing microarray to query Toxoplasma gondii and Hammondia hammondi transcript abundance in sporulated oocysts. Oocysts of the VEG strain of Toxoplasma gondii, and the HhCatGer041 strain of Hammondia hammondi, were isolated from cat feces by sucrose flotation, and sporulated for ~3-6 months in 2% sulfuric acid. RNA was isolated from Bleach-treated oocyst preparations using the Trizol reagent. RNA was biotinylated for hybridization to Toxo 169 Affymetrix chips using the Illumina Total Prep RNA labeling kit (Ambion). For each species 3 separate RNA isolations were performed on the same batch of oocysts and hybridized to individual microarrays.
Project description:Understanding of protein expression difference in the same stage of T.gondii among different genotypes will facilitate the elucidation of genotypic divergence among the T.gondii strains. A 4-plex iTRAQ (isobaric tag for relative and absolute quantitation) based LC-MS/MS approach was employed to survey the differentially expressed proteins of sporulated oocysts between ToxoDB#1 (PRU) strain and ToxoDB#9 (PYS) strain for the first time.
Project description:Eimeria are obligate intracellular protozoan parasites which can affect chickens. After exposure to Eimeria chickens establish (partial) protective immunity to the homologues strain. In this paper we investigate the process responsible for Eimeria protection. In order to find host reactions specificly involved in protection to homologous re-infection we investigated the host reactions after primary infection and a homologous or heterologous secondary infection.<br><br>Broilers were mock infected or infected with E.maxima (Max) at one week of age. Two weeks later broilers were mock infected, infected with E.maxima or E.acervulina. Oocyst output, T-cell population and cytokine mRNA expression profiles and Eimeria DNA profiles were measured 2, 4 and 7 days pi. Specific regulation of gene expression profiles was monitored by a whole genome oligo-array containing 20.673 oligoﾴs at 8 and 24 hours pi.<br><br>