Analysis of Curdlan Production in Agrobacterium sp. ATCC 31749
ABSTRACT: Gene Expression Analysis of Curdlan Production in Agrobacterium sp. ATCC 31749 Two conditions are compared with four biological replicates of each condition, corresponding to eight total samples. The control condition was sampled at 22 hours during the exponential growth phase when no curdlan is produced. The second condition was sampled at 70 hours after the initiation of curdlan production, corresponding to approximately 100 hours.
Project description:This study provides the dectin-1 and NFAT responsive genes for 2h and 4h of curdlan treatment. Affymetrix Mouse Gene 1.0ST Arrays were used to profile gene expression in curdlan-stimulated D1 cells (2h or 4h exposure) that had been treated or not with FK506 for the duration of culture. Dendritic cell line D1 was treated with curdlan for 2h and 4h, and curdlan + Fk506 for 2h and 4h in three replicate.
Project description:Human monocyte derived dendritic cells were exposed to the pure Dectin-1 agonist Curdlan to analyze the early transcriptional activation of DCs. This experiment is related to E-MTAB-750.
Project description:Bone marrow derived macrophages from wt and card9 KO mice were stimulated with CpG, Curdlan or TDB, an analogon to the mycobacterial cord factor TDM for 48h, respectively. Keywords: card9 knockout, macropphage, TDB wt or card9 KO macrophages stimulated for 48h
Project description:A simultaneous engagement of different pathogen recognition receptors provides a tailor made adaptive immunity for an efficient defence against distinct pathogens. For example, cross talk of TLR and c-type lectin signalling effectively shapes distinct gene expression patterns by integrating the signals at the level of NF-κB. Here, we extend this principle to a strong synergism between the Dectin-1 agonist, curdlan, and an inflammatory growth factor, GM-CSF. Both together act in synergy in inducing a strong inflammatory signature which converts immature DCs to potent effector DCs. A variety of cytokines (IL-1β, IL-6, TNF-α, IL-2 and IL-12p70), costimulatory molecules (CD80, CD86, CD40 and CD70), chemokines (CxCl1, CxCl2, CxCl3, CCl12, CCl17) as well as receptors and molecules involved in fugal recognition and immunity such as Mincle, Dectin-1, Dectin-2 and Pentraxin 3 are strongly up-regulated in DC treated simultaneously with curdlan and GM-CSF. The synergistic effect of both stimuli resulted in strong IKBα phosphorylation, in its rapid degradation and in enhanced nuclear translocation of all NF-κB subunits. We further identified MAPK ERK, as one possible integration site of both signals, since its phosphorylation was clearly augmented when curdlan was co-applied with GM-CSF. Our data demonstrate that the immunomodulatory activity of curdlan requires an additional signal provided by GM-CSF to successfully initiate a robust β-glucan specific cytokine and chemokine response. The integration of both signals clearly prime and tailor a more effective innate and adaptive response against invading microbes and fungi. CD11b+ fraction of FLT3L generated BM DCs (3-4 x106) were stimulated for 4 hours with 100 or 1 μg/ml curdlan in presence or absence of 5 ng/ml GM-CSF in triplicates.
Project description:We performed gene expression microarray experiments to compare the global transcriptional response induced by b-glucans and LPS with their secretomes. We identified 1683, 767 and 1447 genes with over two-fold increase or decrease in LPS-, curdlan- or GBY-stimulated macrophages, respectively. We show that both LPS and b-glucan induces significant gene expression changes in macrophages, but only b-glucans activate a robust protein secretion. The gene expression of human primary macrophage cells was measured at 6 hours after exposure the cells to 1 µg LPS or 10 µg Curdlan or 100 µg GBY (glucan from baker´s yeast), also 0-group was included. Three independent experiments were performed using three different donors for each experiment.
Project description:In order to dissect the response from different fungal cell wall components, monocyte-derived dendritic cells obtained from healthy donors were collected and either left untreated, or treated for four hours with either mannan, zymosan, curdlan, whole yeast, or yeast spores. This experiment is related to E-MTAB-1213.
Project description:Gene Expression Analysis of Curdlan Production in Agrobacterium sp. ATCC 31749 Overall design: Two conditions are compared with four biological replicates of each condition, corresponding to eight total samples. The control condition was sampled at 22 hours during the exponential growth phase when no curdlan is produced. The second condition was sampled at 70 hours after the initiation of curdlan production, corresponding to approximately 100 hours.
Project description:Dendritic cells (DCs) are crucial for sensing pathogens and triggering immune response. GM-CSF myeloid dendritic cells (GM-DCs) secrete several cytokines including IL-2 upon activation by pathogen associated molecular pattern (PAMP) ligands. DC IL-2 has been shown to be important for innate and adaptive immune responses, however its importance in DC physiology has never been demonstrated. This is due to ambiguity in expression of the CD122 subunit of the IL-2 trimeric receptor complex crucial for signaling. We show here that autocrine IL-2 signaling is functional in GM-DCs in early time window of stimulation with PAMPs. IL-2 signaling selectively activates the JAK/STAT5 pathway by assembling holo-receptor complexs at the cell surface. Autocrine IL-2 signaling inhibits survival of PAMP matured GM-DCs which is crucial for maintaining immune tolerance and preventing autoimmunity. Our findings suggest immune regulation by a novel autocrine signaling pathway that can potentially be exploited in DC immunotherapy. Microarray technology was used to understand the role of IL-2 signaling in DC. Microarray was performed to investigate the role of IL-2 signaling in DC physiology. Hence Wt or IL-2-/- BMDCs were either treated or not with curdlan for 6h and taken for microarray.
Project description:Alzheimer's disease (AD) is the most common form of dementia, characterized by progressive cognitive impairment and neurodegeneration as a result of abnormal neuronal loss. To elucidate the molecular systems associated with AD, we characterized the gene expression changes associated with multiple clinical and neuropathological traits in 1,053 postmortem brain samples across 19 brain regions from 125 persons dying with varying severities of dementia and variable AD-neuropathology severities. 125 human brains were accessed from the Mount Sinai/JJ Peters VA Medical Center Brain Bank (MSBB). This brain resource was assembled after applying stringent inclusion/exclusion criteria and represents the full spectrum of clinical and neuropathological disease severity in the absence of discernable non-AD neuropathology. RNA samples from 19 brain regions isolated from the 125 MSBB specimens were collected and profiled using Affymetrix Genechip microarrays. There were 50 to 60 subjects per brain region with varying degrees of AD pathological abnormalities.
Project description:Purpose:first,we want to find the genes revelant to curdlan synthesis and oxygen regulation, second, we want to research the function of fnrN gene in Agrobacterium sp. ATCC 31749. Method: samples of cell growth phase, curdlan-producing phase (normoxia) and curdlan-producing phase (micro-oxygen treated) in both Agrobacterium sp. ATCC 31749 wild strain and ΔfnrN strain were collectecd to extract mRNA. Each sample was treated in duplicate. The softwares we used include fastqc, trimmomatic, TopHat2 and Cufflinks. Illumina Hiseq4000 was used to complete the research. Overall design: 12 samples: samples of cell growth phase, curdlan-producing phase (normoxia) and curdlan-producing phase (micro-oxygen treated) in both Agrobacterium sp. ATCC 31749 wild strain and ΔfnrN strain were collectecd to extract mRNA. Each sample was treated in duplicate.