Project description:Transcriptional profiling of Candida albicans after 3 h phagocytosis by vehicle DMSO-treated macrophages (intact, expanding phagosomes) or calcium chelator BAPTA-AM-treated macrophages (inhibits lysosomal repair of expanding phagosomes, leading to phagosome rupture) to determine the effect of preventing phagosome expansion on C. albicans gene expression after phagocytosis by macrophages. Cultivation of Candida only for 3 h in DMEM-FBS cell culture medium or YPD complex medium as non-phagocytosis control conditions.
Project description:To investigate the effect of progranulin on macrophages phagocytosis and killing of Candida albicans, we designed RNA-Seq analysis of wild-type and PGRN-/- macrophages challenged with Candida albicans in vitro at one time point.
Project description:The effects of the SCFA crotonate on fungal and host transcriptomes were addressed, following infection of mouse bone marrow-derived macrophages(BMDMs) with Candida albicans.
Project description:Purpose: The purpose of this study was to simulataneously examine the host and fungal pathogen transcriptional profiles of four distinct infection fates during macrophage and Candida albicans interactions Methods: Membrane stained (Deep Red),primary, bone marrow derived, murine macrophages and Candida albicans expressing GFP and mCherry were exposed to each other over a four hour time course. Samples were collected at 0, 1, 2 and 4 hours and sorted for four infection subpopulations: 1. Macrophages which phagocytosed live C. albicans (GFP+ /mCherry+ /Deep Red +), 2. Macrophages which phagocytosed dead C. albicans (GFP- /mCherry+ /Deep Red +), 3. Uninfected macrophages(GFP- /mCherry- /Deep Red +) and 4. Unengulfed C. albicans (GFP+ /mCherry + /Deep Red -). Unexposed controls were also collected for some time points (i.e. macrophages never exposed to C. albicans and C. albicans never exposed to macrophages). Single macrophages infected with live or dead C. albicans were also sorted. Smart-seq2 was used to create libraries for both infection subpopulation and single, infected cell samples that were sequenced on Illumina’s Miseqand Nextseq. Basic quality assessment of Illumina reads and sample demultiplexing was done with Picard version 1.107 and Trimmomatic. Samples profiling exclusively the mouse transcriptional response were aligned to the mouse transcriptome generated from the v. Dec. 2011 GRCm38/mm10 and a collection of mouse rRNA sequences from the UCSC genome website. Samples profiling exclusively the yeast transcriptional response were aligned to the C. albicans transcriptome strain SC5314 version A21-s02-m09-r10 downloaded from Candida Genome Database. Samples containing both macrophages and C. albicans were aligned to a “composite transcriptome” made by combining the mouse transcriptome and C. albicans transcriptomes described above and alignment was done via BWA (version 0.7.10-r789.) Multi-reads (reads that aligned to both host and pathogen transcripts) were discarded. Then, each host or pathogen sample file were aligned to its corresponding reference using Bowtie2 and RSEM (v.1.2.21). Transcript abundance was estimated using transcripts per million (TPM). For subpopulation samples, TPM was calculated using edgeR, all as implemented in the Trinity package version 2.1.. Genes were considered differentially expressed only if they had a 4-fold change difference (> 4 FC) in TPM values and a false discovery rate below or equal to 0.001 (FDR < 0.001), unless specified otherwise. For single macrophages infected with C. albicans, samples were aligned to the combined transcriptome as described above and RSEM was used to calculate TPM. Results: We were able to simultaneously measure the host and fungal pathogen transcriptional profiles of four distinct infection fates during macrophage and Candida albicans interactions Conclusions: Our study represents an analysis of both distinct infection populations of macrophages and fungus.
Project description:To investigate the effect of Progranulin on neutrophils phagocytosis and killing of Candida albicans, we designed RNA-Seq analysis of wild-type and PGRN-/- neutrophils challenged with Candida albicans in vitro at one time point.
Project description:This SuperSeries is composed of the following subset Series: GSE24073: Transcriptional profile of Candida albicans during Hypoxic conditions. GSE24074: Transcriptional profile of Candida albicans DAY286 culture without ketoconazole versus DAY286 culture with 0.04 μg/ml ketoconazole, both at 20% oxygen (normoxia). GSE24075: Transcriptional profile of Candida albicans DAY286 versus UPC2 delete, both at 1% oxygen (hypoxia). Refer to individual Series
Project description:Transcriptional profiling of Candida albicans SC5314 comparing C. albicans grown in RPMI1640 or in RPMI1640 with 100ug/ml AAT. Goal was to determine the effects of AAT on global C. albicans gene expression.