Project description:A first generation Affymetrix GeneChip® Porcine genome array was used to profile the gene expression in porcine mesenteric lymph nodes over a time course of infection with S. Typhimurium, including the acute (8 hours post inoculation (hpi), 24 hpi, 48 hpi) and chronic (21 days post-inoculation (dpi)) stages of infection. Our objectives were to 1) identify and examine the stereotypical gene expression response within host MLN to S. Typhimurium infection, 2) characterize global host responses by revealing the specific features of the hostâs innate immunity pathways, and 3) explore if and how S. Typhimurium may escape the host immune response and develop into a carrier state. Our study has attempted to investigate the features of host gene expression profiling during S. Typhimurium infection at the acute and chronic infection stages and to explore the mechanism by which S. Typhimurium can escape from the host immune response and develop a carrier state in the host. In conclusion, by using the Affymetrix porcine GeneChip, 848 differentially expressed genes were identified in porcine MLN during infection and several specific features of host response were revealed by gene cluster and pathway analysis. Our data are the first report to investigate global host responses to S. Typhimurium in porcine MLN, and this new study provides data applicable for studying enteric salmonellosis of pigs, as well as humans. Experiment Overall Design: Fifteen piglets from Salmonella spp.-free sows were weaned at 10 days (d) of age, shipped to the National Animal Disease Center, Ames, IA and raised in isolation facilities. To confirm that all piglets were free of Salmonella spp. prior to challenge, bacteriological cultures were performed on rectal swabs twice. Seven week old pigs were randomly divided into 2 groups, 3 non-infected pigs and 12 infected pigs. Three non-infected control pigs were necropsied 2 days prior to experimental infection. On day 0, pigs in the infected groups were intranasally challenged with 1 billion CFU of Salmonella enterica serovar Typhimurium. Three infected pigs were necropsied at 8 hours post-inoculation (hpi), 24 hpi, 48 hpi and 21 days post-inoculation (dpi). Tissue samples from the mesenteric lymph nodes (MLN) were collected and immediately frozen in liquid nitrogen for RNA isolation.
Project description:To better understand how Tritrichomonas arnold colonization impacts the reovirus-mediated proinflammatory response in mesenteric lymph nodes, we examined the transcriptional profile of mesenteric lymph nodes For RNA-sequencing single cell suspension of mesenteric lymph nodes were lysed in RLT buffer (Qiagen) and RNA was isolated
Project description:To understand the host transcriptional response to S. enterica serovar Choleraesuis (S. Choleraesuis), the first generation Affymetrix porcine GeneChip® was used to identify differentially expressed genes in the mesenteric lymph nodes responding to infection at acute (8 hours (h), 24h, 48h post-inoculation (pi)) and chronic stages (21 days (d) pi); The objectives of this study were to identify and examine the stereotypical gene expression response within the host mesenteric lymph nodes to S. Choleraesuis infection, and to characterize the global host responses by revealing the specific features of the hostâs innate immunity. Experiment Overall Design: Fifteen piglets from Salmonella spp.-free sows were weaned at 10 days (d) of age, shipped to the National Animal Disease Center, Ames, IA and raised in isolation facilities. To confirm that all piglets were free of Salmonella spp. prior to challenge, bacteriological cultures were performed on rectal swabs twice. Seven week old pigs were randomly divided into 2 groups, 3 non-infected pigs and 12 infected pigs. Three non-infected control pigs were necropsied 3 days prior to experimental infection. On day 0, pigs in the infected groups were intranasally challenged with 1 billion CFU of Salmonella enterica serotype Choleraesuis x3246. Three infected pigs were necropsied at 8 hours post-inoculation (hpi), 24 hpi, 48 hpi and 21 day post-inoculation (dpi). Tissue samples from the mesenteric lymph nodes (MLN) were collected and immediately frozen in liquid nitrogen for RNA isolation.
Project description:To fully quantify differences in the lymph composition and associated dendritic cells antigenic load, from different anatomical districts and in physiological and pathological conditions, we collected the afferent lymph draining to the cervical and mesenteric lymph nodes in healthy mice. Most of the proteome present in the mesenteric afferent lymph, showed a profile of proteins involved in different metabolic pathways associated with lipoproteintransport, and lipid metabolism, such as adipocyte-type fatty acid binding protein, Apolipoproteins A, B, C and E, and phospholipid transfer proteins, consistent with the known role of the mesenteric lymph in chylomicron transport. Network analysis on the mesenteric afferent lymph unique/enriched proteome highlighted pathways associated with lipase and hydrolase activity, lipoproteins remodeling, fat digestion and absorption, triglyceride catabolism and gut-associated immune cells and cytokines responses. Among the proteome shared across tissue a brain-specific or highly enriched proteome including glia maturation factor, nerve growth factor, mesencephalic astrocyte-derived neurotrophic factor, alpha-crystallin, brain-specific isoform of glycogen phosphorylase, and proteins associated with voltage-dependent channels, were uniquely observed in the lymph harvested from the afferent lymphatics entering the deep cervical nodes. Network analysis on the afferent cervical unique/enriched proteome highlighted pathways associated with neurotransmitter release cycle, synaptic transmission, neuronal development, mitochondrial activity, and an overall CNS proteome.