Project description:Type I strains of Toxoplasma gondii exhibit phenotypic variation, but it is uncertain how differently type I strains modulate the host cell. We determined differential host modulation by type I strains through microarray.
Project description:Toxoplasma strains have been shown to modulate host cell transcription. We have found a type II Toxoplasma gene, GRA15, which activates the nuclear translocation of the NF-kappaB p65 transcription factor. We used microarrays to determine how GRA15II modulates host cell transcription.
Project description:Through targeted CRISPR screening of the Toxoplasma gondii secretome, we identified GRA57 as a novel secreted effector required for survival in interferon-gamma (IFNg)-activated human foreskin fibroblasts (HFFs). In this experiment we aimed to determine if GRA57 affects the host cell transcriptional response to Toxoplasma, and also whether deletion of GRA57 affects the parasite transcriptome.
Project description:Type I strains of Toxoplasma gondii exhibit phenotypic variation, but it is uncertain how differently type I strains modulate the host cell. We determined differential host modulation by type I strains through microarray. HFFs were infected with RH-ERP, RH-JSR and GT1 for 24 hours. Total RNA was isolated and hybridized to Affymetrix GeneChip Human Genome U133A 2.0 arrays.
Project description:Toxoplasma strains have been shown to modulate host cell transcription. We have found a type II Toxoplasma gene, GRA15, which activates the nuclear translocation of the NF-kappaB p65 transcription factor. We used microarrays to determine how GRA15II modulates host cell transcription. HFFs were infected with type I (RH), type I GRA15II (RH GRA15II), type II (Pru), type II GRA15KO (Pru GRA15KO), type III (cep), or type III GRA15II (cep GRA15II) parasites for 18-24 hours. Some samples were also stimulated with TNF-alpha. Total RNA was isolated and hybridized to Affymetrix GeneChip Human Genome U133A 2.0 arrays.
Project description:Pre-infection with Toxoplasma parasites inhibits induction of an interferon gamma responsive gene expression program. In this study we aimed to determine what role host negative regulatory proteins that are induced by Toxoplasma infection, such as SOCS-1, have in this inhibition. We used Affymetrix microarrays to analyze host cell transcription after pre-treatment with the protein synthesis inhibitor cyclohexamide (CHX), Toxoplasma infection, and subsequent interferon-gamma stimulation.
Project description:Pre-infection with Toxoplasma parasites inhibits induction of an interferon gamma responsive gene expression program. In this study we aimed to determine what role host negative regulatory proteins that are induced by Toxoplasma infection, such as SOCS-1, have in this inhibition. We used Affymetrix microarrays to analyze host cell transcription after pre-treatment with the protein synthesis inhibitor cyclohexamide (CHX), Toxoplasma infection, and subsequent interferon-gamma stimulation. Human foreskin fibroblasts (HFFs) were left untreated or treated with CHX for 40min. Cells were infected, or not, with type I (RH) Toxoplasma parasites, for one hour, and subsequently stimulated with human IFN-γ for one hour. Total RNA was isolated and hybridized to Affymetrix Human U133A 2.0 gene chips.