Avian duodenum tissues: primary vs. secondary E. acervulina infected
ABSTRACT: Global transcriptional responses in duodenal intestinal epithelia of chickens following primary and secondary Eimeria acervulina infections. Simple loop hybridization (day 0 vs. days 1-2, days 1-2 vs. 3-4, days 3-4 vs. 5-6, and days 5-6 vs. 7-8) per primary or secondary infections.
Project description:Relative expression levels of mRNAs in chicken IEL experimentally infected with EA, EM, or ET were measured at 1 to 6 days post-infection (dpi) following primary and secondary infections. One week-old chickens were uninfected (negative control) or were orally inoculated with sporulated oocysts of EA, EM, or ET. One week later, the infected chickens were challenged with an identical inoculum of the homologous parasite. Intestinal samples were collected daily from 5 birds in a treatment group at from 1 to 6 dpi following primary and secondary infections. Cecum, duodenum, and jejunum were collected from the birds challenged with E. acervulina, E. maxima, and E. tenella, respectively. Uninfected control samples and one of the 3 infection group samples were labeled with different fluorescent dyes and hybridized simultaneously on the same slide using a reference design with a dye swap protocol. Thirty seven-condition experiment, Non-infected control vs. Primary or secondary EA, EM, or ET infected IEL at 1 to 6. Biological replicates: 2 replicates with dye-switching from each infection groups. Two replicates per array.
Project description:This study investigated the ability of two novel adjuvant formulations, QCDC (Quil A/cholesterol/DDA/Carbopol) and QCDCR (QCDC/Bay R1005), in combination with a recombinant profilin vaccine, to modulate host protective immunity and to alter new gene expression during experimental avian coccidiosis. Four-condition experiment, Profilin only vs. Non-vaccination, Profilin only vs. Co-vaccination of QCDC plus profilin, Profilin only vs. Co-vaccination of QCDCR plus profilin, Biological replicates: 2 profilin only replicates, 2 Non-vaccination replicates, 2 QCDC plus profilin replicates, 2 QCDCR plus profilin replicates with dye-switching.
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>
Project description:Relative expression levels of mRNAs in chicken cecal epithelia experimentally infected with Eimeria tenella were measured at 4.5 days post-infection. Two weeks old chickens were uninfected (negative control) or were orally inoculated with sporulated oocysts of Eimeria tenella. Cecal epithelia samples were collected from >12 birds in infected or uninfected group at 4.5 d following infections, in which samples from 4 birds were pooled together to form a total 3 biological replicates in each group. Parasite merozoites were also collected from four infected chickens at 5 d after infections. Uninfected control samples, merozoites and infection group samples were selected for RNA extraction and hybridization on Affymetrix microarrays. We used Affymetrix GeneChip chicken genome arrays to detail the chicken cecal epithelia gene expression in the control and E. tenella-infected birds. Infected, uninfected chicken cecal epithelia and merozoites were selected for RNA extraction and hybridization with Affymetrix microarrays. Our goal was to analyze global transcriptome changes in chicken cecal mucous membranes in response to E. tenella infection in vivo. We used infected (T1,T2,T3; three biological replicates) and uninfected (Neg1, Neg2, Neg3; three biological replicates) samples to identify genes that were differentially expressed. Meanwhile, RNA and probes were also prepared from parasite merozoites (Mzt) from infected samples (Mzt) and used as an additional control in microarray hybridization.
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: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:Many eukaryotic developmental and cell fate decisions are effected post-transcriptionally that mechanistically involve RNA binding proteins as regulators of translation of key mRNAs. In the unicellular eukaryote malaria parasite, Plasmodium, one of the most dramatic changes in cell morphology and function occurs during transmission between mosquito and human host. In the mosquito salivary glands, Plasmodium sporozoites are slender, motile and remain infectious for several weeks; only after transmission and liver cell invasion, does the parasite rapidly transform into a round, non-motile exo-erythrocytic form (EEF) that gives rise to thousands of infectious merozoites to be released into the blood stream. Here we demonstrate a Plasmodium homolog of the RNA binding protein, Pumilio, as a key regulator of the sporozoite to EEF transformation. In the absence of Pumilio-2 (Puf2) Plasmodium berghei sporozoites initiate early stage EEF development inside mosquito salivary glands with characteristic morphological changes; puf2- salivary gland sporozoites lose gliding motility, cell traversal ability and are less infective. Global expression profiling confirmed that transgenic parasites exhibit genome-wide transcriptional adaptations typical for Plasmodium intra-hepatic development. The data demonstrate that Puf2 is a key player in regulating developmental control, and imply that transformation of salivary gland-resident sporozoites into early liver stage parasites is regulated by a post-translational mechanism.
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:Highly pathogenic avian influenza virus (HPAIV) is a permanent threat due to its capacity to cross species barriers and generate severe infections and high mortality in humans. Recent findings have highlighted the potential role of PB1-F2, a small accessory influenza protein, in the pathogenesis process mediated by HPAIV in mammals. In this study, using a recombinant H5N1 HPAIV (wt) and its PB1-F2-deleted mutant (ΔF2), we studied the effects of PB1-F2 in a chicken model. Unexpectedly, when using low inoculation dose we observed that the wt-infected chickens had a higher survival rate than the ΔF2-infected chickens, a feature that contrasts with what is usually observed in mammals. High inoculation dose had similar mortality rate for both viruses, and comparison of the bio-distribution of the two viruses indicated that the expression of PB1-F2 allows a better spreading of the virus within chicken embryos. Transcriptomic profiles of lungs and blood cells were characterized at two days post-infection in chickens inoculated with the wild type (wt) or the ΔF2 mutant viruses. In lungs, the expression of PB1-F2 during the infection induced pathways related to calcium signaling and repressed a large panel of immunological functions. In blood cells, PB1-F2 was associated to a gene signature specific for mitochondrial dysfunction and down-modulated leucocytes activation. Finally we compared the effect of PB1-F2 in lungs of chickens and mice. We identified that gene signature associated to tissue damages is a PB1-F2 feature shared by the two species; by contrast, the early inhibition of immune response mediated by PB1-F2 observed in chickens is not seen in mice. In summary, our data suggest that PB1-F2 expression deeply affect the immune host response in chickens in a way that may attenuate pathogenicity, a feature differing from what was previously observed in mammal species. Three-condition experiment, virus-infected (wt or ΔF2) vs. Mock-infected chickens. Biological replicates: 2x5 control replicates, 5 wt replicates, 5 ΔF2 replicates.