Project description:LGG exposure of C. elegans protects C. elegans against pathogen infection and prolongs lifespan. In particular, it prolongs lifespan by up-regulating specific genes to pathogenic microorganisms. We used microarrays to elucidate miRNA expression to determine how LGG exposure in C. elegans affected miRNAs and identified miRNAs that were significantly regulated in this process.
Project description:YAMC (young adult mouse colon) cells are a conditionally immortalized mouse colonic intestinal epithelial cell line derived from the Immortimouse.Prior to each experiment, cells were plated at a density of 7.5 x 105 cells per p100mm dish.Cells were treated with LGG (the probiotic Lactobacillus GG ) conditioned media overnight, then harvested the following day. RNA were extracted with Trizol and purified with Qiagen kit. Affymetrix microarray chip Mouse 430A containing 19,000 murine genes was run in duplicate(experiment on 7-24-03 and 1-22-04 ). Data were analyzed using Affymetrix Genechip Operating Software (GCOS, Version 1.0) In each case; LGG treatment was compared to mock treatment controls. Results are expressed as fold change of treated cells as compared to controls, as calculated using GENESPRING software (Version 4.2.1, Silicon Genetics, Mountain View, CA), Statistical analysis was performed using D chip. DNA microarray experiments demonstrate that epithelial cell hsp70 is one of the most highly upregulated genes in response to LGG-CM treatment Keywords: repeat sample
Project description:Analysis of human primary macrophages after live Lactobacillus rhamnosus GG (LGG) or LC705 stimulation for 6h and 24h. The results reveal novel mechanisms for probiotics-induced activation of the healthy human innate immune system. Macrophages are phagocytic cells of the innate immune system that perform sentinel functions to initiate appropriate responses to surrounding stimuli. Macrophages that reside at mucosa encounter ingested bacteria. Our understanding of the responses elicited by nonpathogenic bacteria in human innate immune system is limited. Lactobacillus are nonpathogenic bacteria commonly used in food and as supplements with health-promoting probiotic potential. In this study, we have utilized global gene expression profiling to compare the responses of human primary macrophages to two closely related, well-characterized L. rhamnosus strains LGG and LC705. Our results demonstrate that live LGG and LC705 induced quantitatively different gene expression in macrophages. A gene ontology analysis revealed functional similarities and differences in responses to LGG and LC705. Both LGG and LC705 induced IL-1b production in macrophages that required caspase-1 activity. LC705 but not LGG induced a strong type I IFN-dependent gene activation that correlated with the ability of LC705 to prevent influenza A virus replication and production of viral proteins in macrophages. Differentiated 7d human primary macrophages from 18 healthy individuals were stimulated with either live L. rhamnosus GG (LGG) or LC705 for 6 h and 24 h in RPMI medium containing penicillin and streptomycin. As a control, macrophages were stimulated with the medium only. The experiment was performed three times, each time with cells from six different individuals (samples 1-3). For each experiment, macrophages were stimulated separately as described, and macrophages from different donors were pooled after each stimulation experiment. Extracted RNA was hybridized to AffymetrixM-BM-. U133-plus2.0 GeneChipM-BM-. arrays.
Project description:Low-grade glioma (LGG) is a primary, slow-growing brain tumor; however, its treatment and prognosis remain challenging. In this study, we analyzed cancer data from the TCGA database, focusing particularly on the expression of the CDKN3 gene in LGG. The results showed that high CDKN3 expression in LGG patients was significantly associated with poor survival outcomes. Further gene expression analysis revealed that 379 genes were significantly upregulated in LGG samples with high CDKN3 expression, and these genes were primarily involved in the mitotic cell cycle and extracellular matrix organization. Additionally, high CDKN3 expression was closely linked to key signaling pathways such as tumor inflammation, hypoxic response, and tumor proliferation. Immune microenvironment analysis showed that high CDKN3 expression significantly increased the expression of CD4+ T cells and specific immune checkpoint genes, suggesting a potentially poor response to immune checkpoint blockade therapy. Through in vitro and in vivo experiments, we confirmed that CDKN3 silencing significantly inhibited the proliferative capacity of LGG cells. Proteomics revealed that CDKN3 can bind ARG1 and upregulate intracellular arginine concentrations. These findings not only improve our understanding of LGG biology but also provide scientific evidence for developing potential therapeutic strategies targeting CDKN3.
Project description:Low-grade glioma (LGG) is a primary, slow-growing brain tumor; however, its treatment and prognosis remain challenging. In this study, we analyzed cancer data from the TCGA database, focusing particularly on the expression of the CDKN3 gene in LGG. The results showed that high CDKN3 expression in LGG patients was significantly associated with poor survival outcomes. Further gene expression analysis revealed that 379 genes were significantly upregulated in LGG samples with high CDKN3 expression, and these genes were primarily involved in the mitotic cell cycle and extracellular matrix organization. Additionally, high CDKN3 expression was closely linked to key signaling pathways such as tumor inflammation, hypoxic response, and tumor proliferation. Immune microenvironment analysis showed that high CDKN3 expression significantly increased the expression of CD4+ T cells and specific immune checkpoint genes, suggesting a potentially poor response to immune checkpoint blockade therapy. Through in vitro and in vivo experiments, we confirmed that CDKN3 silencing significantly inhibited the proliferative capacity of LGG cells. Proteomics revealed that CDKN3 can bind ARG1 and upregulate intracellular arginine concentrations. These findings not only improve our understanding of LGG biology but also provide scientific evidence for developing potential therapeutic strategies targeting CDKN3.
Project description:Analysis of human primary macrophages after live Lactobacillus rhamnosus GG (LGG) or LC705 stimulation for 6h and 24h. The results reveal novel mechanisms for probiotics-induced activation of the healthy human innate immune system. Macrophages are phagocytic cells of the innate immune system that perform sentinel functions to initiate appropriate responses to surrounding stimuli. Macrophages that reside at mucosa encounter ingested bacteria. Our understanding of the responses elicited by nonpathogenic bacteria in human innate immune system is limited. Lactobacillus are nonpathogenic bacteria commonly used in food and as supplements with health-promoting probiotic potential. In this study, we have utilized global gene expression profiling to compare the responses of human primary macrophages to two closely related, well-characterized L. rhamnosus strains LGG and LC705. Our results demonstrate that live LGG and LC705 induced quantitatively different gene expression in macrophages. A gene ontology analysis revealed functional similarities and differences in responses to LGG and LC705. Both LGG and LC705 induced IL-1b production in macrophages that required caspase-1 activity. LC705 but not LGG induced a strong type I IFN-dependent gene activation that correlated with the ability of LC705 to prevent influenza A virus replication and production of viral proteins in macrophages.
Project description:We treated WT or Myd88-knockout mice bearing MC38 tumors with oral administration of live LGG combined with anti-PD-1 antibody or not . CD11c+ DCs were isolated from MLN and then RNA-seq performed. The goals of this study are to identify the activated cellular pathways in DCs with oral administration of live LGG or the combination treatment. Samples were sequenced on an Illumina Hiseq instrument. The short sequence reads were aligned to mouse mm10 reference genome using STAR (2.4.0) and then assembled and quantified by HTSeq (0.6.1) with Gencode M8 as annotation. DESeq2 was used for differential gene expression analysis among subgroups. Hierarchical clustering and principal component analysis were plotted applying FactoMineR and gplots libraries in R. log2 [FPKM] was calculated for each gene. We found that cytokine-related pathways, response to interferon pathway and T cell activation pathway were activated in combination treatment group and Response to interferon beta pathways was activated when LGG alone treated.