Project description:The highly prevalent gut helminth, Ascaris suum, compromise pigs health and reduce farm productivity worldwide. The closely related human parasite, A. lumbricoides, infects more than 800 million people and causes approximately 1.31 million disability-adjusted life years. The humaninfections are often chronic by nature and the parasites have a profound ability to modulate their hosts immune responses. This study provides the first in-depth characterization of extracellular vesicles (EVs) from different developmental stages and body parts of A. suum and their potential role in the host-parasite interplay. The release of EVs during the third larval stage (L3), L4 and adults was demonstrated by Transmission Electron Microscopy, and the uptake of EVs from adult A. suum in intestinal epithelial cells followed by accumulation of RNA in the nucleus by confocal microscopy. Next Generation Sequencing of EV-derived mi/m?RNA identified a number of micro(mi)RNAs from the different A. suum life stages and body parts and potential transcripts of potential host immune targets, such as IL-13, IL-25 and IL-33, were identified. Proteomics of EVs identified several proteins with immunomodulatory properties and other proteins previously shown to be associated with parasite EVs. Furthermore, EVs from A. suum body fluid stimulated the production of the pro-inflammatory cytokines IL-6 and TNF-α in dendritic cells in vitro. Taken together, these results suggest that A. suum EVs and their cargo may play a role in host-parasite interactions. This knowledge may pave the way for novel strategies for helminth infection control and knowledge of their immune modulatory role.
Project description:Over a billion people are infected by Ascaris spp. intestinal parasites. We investigated gene expression by various tissues of adult A. suum, and used that information to help resolve basic functions of these tissues, basic biology of parasitism and functional aspects of nematode genomes. The A. suum genome was sequenced and assembled to allow generation of microarray elements. Expression of over 40,000 60-mer elements was investigated in a variety of tissues from both male and female adult worms. Nearly 50 percent of the elements for which signal was detected exhibited differential expression among different tissues. The unique profile of transcripts identified for each tissue clarified functional distinctions among tissues, such as chitin binding in the ovary and peptidase activity in the intestines. Interestingly, hundreds of gender specific elements characterized female or male intestinal tissues, respectively. A. suum genes from the same family were frequently expressed differently among tissues. Transcript abundance for genes specific to A. suum, by comparison to Caenorhabditis elegans, varied to a greater extent among tissues than for genes conserved between A. suum and C. elegans. Analysis using C. elegans protein interaction data identified functional modules conserved between these two nematodes, resulting in identification of functional predictions of essential subnetworks of protein interactions and how these networks may vary among nematode tissues. A notable finding was very high module similarity between adult reproductive tissues and intestine. Our data identified gene and predicted protein determinants that distinguish functions of individual adult A. suum tissues as well those that likely serve all the tissues investigated in our study. The size of A. suum adults (30-40 cm compared to less than 1 cm for many nematodes) enabled this resolution and will facilitate experimental dissection of individual and interactive functions of these tissues in this and other parasitic nematodes. 10 samples were prepared from the tissues of adult worms of both genders to be analyzed. Two to three replicates were done for each sample, swapping dyes and channel for the test and the control for each sample. The control consisted of pooled worm tissues from both genders with an equal amount being included from each sample. These samples and the control were hybridized against an Agilent 4x44k format custom chip spotted with 42,212 probes. This probeset was created from 38,768 predicted protein coding sequences from an Ascaris suum genome survey sequence (GSS) project, and also from 5,176 EST contigs that were not covered by this predicted GSS gene-set. These 43,944 putative gene sequences from Ascaris suum were filtered for cross-reaction to non-coding sequences using Agilent eArray, and our final set of 42,212 probes was determined.
Project description:We show here the transcriptional response in the intestine of pigs infected or not with Ascaris suum, fed either a control pr a PAC-enriched diet
Project description:We show here the transcriptional response in the lung of pigs infected or not with Ascaris suum, fed either a control pr a PAC-enriched diet
