Dataset Information

Paired-end RNA Sequencing on primary human umbilical endothelial cells (HUVEC) exposed to uninfected or bacterially-infected PMA-differentiated macrophage-like U937 for 8 or 24 h

ABSTRACT: We sought to identify changes in gene expression that occur when endothelial cells in monolayer interact with either uninfected or Listeria monocytogenes-infected macrophages. To that end we seeded HUVEC in monolayer and either exposed them to nothing, or to uninfected PMA-differentiated macrophage-like U937, or to Listeria monocytogenes-infected PMA-differenitated U937. At 8 or 24 hours post U937 exposure (hpe), we washed the samples thoroughly to remove U937 and then collected the mRNA to to perform RNA-Sequencing and gene expression profiling for the six conidtions described. By analyzing the differentially expressed genes between all conditions we find a significant number of up- or down-regulated genes related to innate immune signaling in both (un)infected U937 exposed HUVEC as compared to control unexposed HUVEC, irrespective of the time post-exposure. For example we found significant upregulation of genes related IL-17, NF-kB and TNF related signaling pathways, though to a lesser degree for HUVEC exposed to uninfected U937 as compared to bacterially infected. Interestingly, we found that only HUVEC exposed to uninfected but not infected U937 and only at 8hpe showed significant upregulation of genes related to focal adhesions and regulation of the actin cytoskeleton. We found that consistent with alterations in the dynamics of HUVEC under this condition, which show a significant rise in the traction on monolayer stresses they exert. At later times post-exposure (i.e., 24 hpe) were these changes in gene expression are not present anymore, HUVEC dynamics are also identical irrespective of whether they were exposed or not to (un)infected U937. Thus the RNA sequnecing results at 8 hpe that reveal upregulation of genes related to focal adhesions and the actin cytoskeleton for uninfected U937 exposed HUVEC match with the results of traction force and monolyaer stress microscopy that reveal strong enhancement of these forces over the first 10 hpe and which are absent in the other two conditions or at the later time point of 24 hpe.

ORGANISM(S): Homo sapiens

PROVIDER: GSE296345 | GEO | 2026/01/27

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other
	GSE296345_Differentially_Expressed_Genes.xlsx	Xlsx
	GSE296345_Differentially_Expressed_Transcripts.xlsx	Xlsx
	GSE296345_Expression_Gene.xlsx	Xlsx
	GSE296345_Expression_Transcript.xlsx	Xlsx

Items per page:

1 - 5 of 5

Similar Datasets

Project description:Purpose: When we infect MDCK epithelial cells we low dosage of Listeria monocytogenes we observe that at late times (24 h) post infection that infected cells pertaining in infection foci get collectively extruded out of the epithelial monolayer forming a 3D mound. The formation of the infection mound results from the competition between two populations: the uninfected cells ("surrounders") that eventually squeeze and extrude the infected cells ("mounders"). Using RNA-Sequencing, we performed gene expression profiling for uninfected MDCK cells, cells infected for 24 h with Listeria monocytogenes at an MOI of 200 (both "mounders" and "surrounders") and cells sorted to infected ("mounders") or uninfected ("surrounders") populations after being infected for 24 h with Listeria monocytogenes. Our goal was to understand how transcriptional regulation leads to the collective extrusion of infected epithelial cells triggered by surrounding uninfected cells. Methods and Results: By analyzing the differentially expressed genes between all conditions we find a significant number of up- or down-regulated genes between all groups. Through pathway enrichemnet analysis we find that compared to cells not exposed to infection "mounders" showed significant downregulation in 31 genes related to tight junctions and ZO-1 was one of the key genes downregulated and to a lesser degree to pathways related to adherens junctions, focal adhesions and regulation of the actin cytoskeleton. "Surrounders" showed significant downregulation to genes related to actin cytoskeleton, to endocytosis to ahderens junctions and to a lesser degree focal adhesions and tight junctions. Especially "surrounders" and to a lesser degree "mounders" showed upregulation on IL-17 and NF-kB and TNF related signaling pathways, which underlines the fact that although "surrounders" are uninfected their distinct mechanical behavior might arise from cytokines released from "mounders" which impact crucially their phenotype. Interestingly "mounders" only showed additionally upregulation on DNA replication, cell cycle related pathways and ferroptosis suggesting that these cells might be undergoing some kind of stress response. Conclusions: Infection of MDCK epithelial cells with Listeria monocytogenes significantly alters the transcriptome of the infected host cells when those are examined 24 h post infection. Both infected and uninfected cells in an infected well show significant transcriptomic changes compared to uninfected cells but also compared to each other. Most importnantly innate immunity related pathways are singificanlty upregulated for un infected cells in an infected well and to a lesser degree for infected cells as compared to cells not exposed to infection.

Project description:Description: Experimental exposure to malaria parasites can lead to development of protective immunity, providing a foothold for the development of a malaria vaccine. The goal of this study is to investigate immune transcriptional profiles associated with malaria protective immune responses induced by experimental Chemo-Prophylaxis and Sporozoites (CPS) immunization of P. falciparum-naive human volunteers. Samples for this study were obtained from a CPS-immunization study conducted in 2011 (ClinicalTrials.gov Identifier: NCT01218893), where healthy volunteers received CPS-immunization with bites from different numbers of P. falciparum-infected mosquitoes (three times 15 (n=5), 10 (n=9) or 5 (n=10)). Five control subjects received uninfected mosquito bites under chloroquine prophylaxis. Fifteen weeks after discontinuation of chloroquine prophylaxis, all volunteers were challenged by exposure to infected mosquito bites. Study Design: A maximum of 30 volunteers were divided into four groups. All volunteers received weekly chloroquine prophylaxis for a period of 13 weeks (91 days). During these 13 weeks, on days 8, 36 and 64 they were exposed to the bites of 15 mosquitoes. Group 1 (n=5, positive control) received three CPS immunizations by 15 mosquitoes infected with P. falciparum sporozoites. Group 2 (n=10) received three times 10 bites from infected mosquitoes and 5 bites from uninfected mosquitoes. Group 3 (n=10) received three times 5 bites from infected mosquitoes and 10 bites from uninfected mosquitoes. Group 4 (n=5), the negative control, received three placebo immunizations with 15 bites of uninfected mosquitoes. Fifteen weeks after discontinuation of chloroquine prophylaxis, all 30 volunteers were challenged at day 196 by the bites of 5 infectious mosquitoes and followed for 21 days. All subjects were treated with chloroquine 21 days after challenge or whenever they had a positive thick smear during that period.

Dataset Information

Paired-end RNA Sequencing on primary human umbilical endothelial cells (HUVEC) exposed to uninfected or bacterially-infected PMA-differentiated macrophage-like U937 for 8 or 24 h

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets