Project description:The Toxoplasma type I ROP16 kinase directly activates the host STAT3 and STAT6 transcription factors and when transgenically expressed in the orally virulent type II strain, promotes host resistance to oral challenge. The transcriptional profile of type II and II+ROP16I infected Peyer's patches and intestines from orally infected mice on day 5 was determined to elucidate host signaling pathways and molecular gene targets that correlate with protective immunity in the gut of orally challenged animals C57BL/6 female mice were orally gavaged with 1000 tissue cysts of the type II or II+ROP16I strain. On Day 5 of infection the Peyer's patches and intestines were analyzed for parasite burden by bioluminescence imaging. Individual Peyer's patches and intestinal sections, corresponding to the middle third of the intestine (~jejunum), that had equivalent parasite burden were snap frozen in liquid nitrogen. The samples were crushed using a sterile 16 G needle in tubes on dry ice. In the case of the intestine, the sample was resuspended in TRizol, extruded through a needle using a syringe and RNA was isolated according to manufacturer's protocol; for the Peyer's patch, the sample was suspended in cell lysis buffer and RNA isolated using the RNeasey kit (Invitrogen) according to the manufacter's protocol.
Project description:The Toxoplasma type I ROP16 kinase directly activates the host STAT3 and STAT6 transcription factors and when transgenically expressed in the orally virulent type II strain, promotes host resistance to oral challenge. The transcriptional profile of type II and II+ROP16I infected Peyer's patches and intestines from orally infected mice on day 5 was determined to elucidate host signaling pathways and molecular gene targets that correlate with protective immunity in the gut of orally challenged animals
Project description:To establish better understanding of cells found in jejunal and ileal Peyer's patches of pigs, we utilized single-cell RNA sequencing scRNA-seq and spatial transcriptomics to recover and analyze cells and spatial regions from sections of jejunum and ileum containing Peyer's patches. Cells identified via single-cell RNA sequencing included B, T/innate lymphoid cell, myeloid, epithelial, and stromal lineage cells. Spatial dots recovered via spatial transcriptomics belonged to regions including villi, crypts, interfollicular/parafollicular zones, follicles, and muscularis. Overall, results provide new information on regional localization and transcriptional profiles of cells in the pig small intestine.
Project description:To gain a comprehensive view of the host response to pathogens within these tissues, we determined the transcriptional profiles of intestinal lymphatic tissue infected with Y. enterocolitica. Expression analysis using Affymetrix GeneChips revealed a complex host response in the Peyers patches (PP) and mesenteric lymph nodes (MLN) following oral infection with Y. enterocolitica. Peyer's patches and mesenteric lymph nodes were surgically removed from uninfected (control) and infected (experimental) mice at 12 hours, day 2, 4 and 7 post oral infection. Tissue samples from 10 mice per time-point were combined. Total RNA was extracted using the Trizol method prior to total cDNA syntesis, labeling and hybridization to Affymetrix MGU74Av2 GeneChips. Data was processed and analyzed using MAS 5.0 and custom analysis protocols. Samples from Peyer's patches and mesenteric lymph nodes are included in this series.
Project description:We have previously shown that aging affects the functional maturation of M cells in the follcile associated epithelium (FAE) of Peyer's patches in the small intestine. To further characterise how aging affects the FAE, mRNA-seq was performed on micro-dissected FAE and villous epithelial sheets as well as whole ileum from young (4 month old) and aged (26-28 month old) mice
Project description:RANKL (receptor acrivator of NFkB ligand) is a member of TNF superfamily cytokines. In the gastrointestinal tract, RANKL is expressed in the stromal cells of Peyer's patches, and involved in the development of the specialized intestinal epithelial cells, called M cells. To identify the genes involved in M-cell development, we treated BALB/c mice with recombinant GST-RANKL. After RANKL-treatment, epithelial cells were isolated from small intestine, and used for microarray analysis. Total of 15 samples were analyzed. We generated the Excel sheet for comparing gene expression.
Project description:RANKL (receptor acrivator of NFkB ligand) is a member of TNF superfamily cytokines. In the gastrointestinal tract, RANKL is expressed in the stromal cells of Peyer's patches, and involved in the development of the specialized intestinal epithelial cells, called M cells. To identify the genes involved in M-cell development, we treated BALB/c mice with recombinant GST-RANKL. After RANKL-treatment, epithelial cells were isolated from small intestine, and used for microarray analysis.
Project description:This study couples a swine experimental model of non-typhoid salmonellosis to laser capture microdissection and microarray analysis to dissect the mechanisms carried out in the follicles of ileum Peyer’s Patches in response to Salmonella Typhimurium infection. Affymetrix GeneChip® Porcine genome array was used to study the gene expression profiles of Peyer's Patches grom control and infected pigs at acute phase of infection.
Project description:Analysis of Ly6Chi monocytes from small intestine lamina propria (SILP) and blood of day 8 Toxoplasma gondii infected mice at gene expression level. The hypothesis tested in the present study was that Ly6Chi monocytes from SILP have altered expression of regulatory factors to blood monocytes. Results provide important information on the regulatory to effector balance of genes expressed by Ly6Chi monocytes during an acute inflammatory response. Ly6Chi inflammatory monocytes were sorted by FACS from the blood or small intestine lamina propria (SILP) of Toxoplasma gondii infected C57BL/6 mice. Cells were isolated at day 8 after infection and total RNA obtained from sorted populations. Three biological replicates were acquired for both blood and SILP from pooled animals.
