Project description:To investigate the transcriptional effects of TgAP2XII-5 on type II Toxoplasma gondii (ME49) in the tachyzoite and bradyzoite stages. Also, to study the transcriptional role of TgAIP1 on ME49 tachyzoites.
Project description:We wanted to determine how type II versus type III Toxoplasma infection affect host gene expression We infected mouse macrophage (RAW264.7 and J774) and dendritic (DC2.4) cell lines with type II (Me49) and type III (CEP)
Project description:We wanted to determine how type II versus type III Toxoplasma infection affect host gene expression We infected mouse macrophage (RAW264.7 and J774) and dendritic (DC2.4) cell lines with type II (Me49) and type III (CEP) cells were grown in T75 until 80% confluency, then infected with parasites for 18 hours at MOI of 7. Then the RNA was harvested from the cells by Trizol.
Project description:Transcriptome analysis of peritoneal lavage of mice infected with T. gondii Toxoplasma gondii is the causative agent of toxoplasmosis in human and animals. In mouse model, T. gondii strains can be divided into three groups, including the virulent, intermediately virulent and non-virulent. The clonal Type I, II and III T. gondii strains belong to these three groups respectively. To better understand the basis of virulence phenotypes, we investigated mouse gene expression responses to the infection of different T. gondii strains at day 5 post intraperitoneal inoculation with 500 tachyzoites. The transcriptomes of mouse peritoneal cells showed that 1927, 1573, and 1009 transcripts were altered more than 2 fold by Type I, II and III infections, respectively, and majority of altered transcripts were shared. Overall transcription patterns were similar in Type I and Type II infections and both had greater changes than that of Type III. Quantification of parasite burden in mouse spleens showed that Type I was 1000 times higher than Type II, and Type II was 20 times higher than Type III. Fluorescence activated cell sorting revealed that Type I and II infections had comparable macrophage populations and both were higher than Type III infection. In addition, Type I infection had higher percentage of neutrophils than that of Type II and III. Taken together, these results suggested that there is a common gene expression response to T. gondii infection in mice. This response is further modified by parasite strain specific factors that determine their distinct virulence phenotypes. We analyzed mRNA from female CD1 outbred mice, 6-8 weeks old infected with Type I, II and III T. gondii strains. We used the Affymetrix Mouse Gene 1.0 ST platform. Raw array data was processed by Partek® Genomics SuiteTM software. Three replicates were performed for Type I-GT1 and Type III-CTG and two replicates for Type II- PTG.
Project description:Toxoplasma gondii is a ubiquitous protozoan pathogen able to infect both mammalian and avian hosts. Surprisingly, just three strains appear to account for the majority of isolates from Europe and N. America. To test the hypothesis that strain divergence might be driven by differences between mammalian and avian response to infection, we examine in vitro strain-dependent host responses in a representative avian host, the chicken. Chicken embryonic fibroblasts were cultivated in vitro and infected with different strains of Toxoplasma gondii (Type II = ME49, Type III = CEP); host transcriptional responses were then analyzed at 24 hours post-infection.
Project description:Previous studies suggested that specific bradyzoite genes were differentially expressed in the three major lineages common to North America and Europe (Radke et al., 2006). In order to determine if these differences extended to the global transcriptome, we characterized whole-cell mRNA levels in both tachyzoite and bradyzoite populations from three primary strain isolates differentiated in vitro. It is well understood in the field that long passage history is associated with the loss of developmental competence (Frenkel et al., 1976). Therefore, the Type I-GT1, Type II-Me49B7 and Type III-CTG strains studied here were maintained at < 20 cell culture passages from the oocyst stage and are capable of completing both intermediate and definitive host life cycles. These experiments used newly constructed Affymetrix GeneChips that include probe sets for ~8000 genes, and we assessed the induction of bradyzoite genes using a strong inducer of bradyzoite differentiation, Compound 1 (Radke et al., 2006). Both tachyzoite and induced (Compound 1/48hrs - bradyzoite) RNA for strains representing the three major lineages of Toxoplasma gondii were hybridized .