Project description:<p>A mechanistic understanding of the health benefits conferred by consumption of probiotic bacteria has been limited by our knowledge of the resident gut microbiota and its interaction with the host. We used fecal samples from a study of twelve healthy individuals aged 65-80 to characterize the structure and functional dynamics of the gut microbiota associated with consumption of the single-organism probiotic, Lactobacillus rhamnosus GG ATCC 53103 (LGG). Samples were collected prior to probiotic consumption (day 0), on day 28 immediately after consuming 10^10 CFU of LGG twice daily for 28 days and day 56, one month after stopping LGG consumption. Our integrative approach incorporated bacterial 16S rRNA gene sequencing, whole-community expression profiling using RNA-seq, and metagenomic sequencing. We highlight the value of combinatorial 'omics methods and concomitant high-resolution informatics to probe the role that probiotics may play on the structure and function of the resident microbiota.</p>
Project description:Lacticaseibacillus rhamnosus GG (LGG) is a widely consumed probiotic whose potential beneficial effects in humans have been examined in over 250 clinical trials. However, the mechanisms by which LGG modulates host gut physiology remains unknown. R. gnavus is a pathobiont that is strongly associated with inflammatory bowel diseases. Germ free mice were mono-associated with L. rhamonosus (LGG) or R. gnavus (RG) for 21 days, ileum bulk RNA-seq were performed to compare the transcriptomic profiles of LGG or RG associated mice with the transcriptome of germ-free mouse ileum.
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
Project description:Lactobacillus rhamnosus GG (LGG) is the most widely used probiotic, but the mechanisms underlying its beneficial effects remain unresolved. Previous studies typically inoculated LGG in hosts with established gut microbiota, limiting the understanding of specific impacts of LGG on host due to numerous interactions among LGG, commensal microbes, and the host. There has been a scarcity of studies that used gnotobiotic animals to elucidate LGG-host interaction, in particular for gaining specific insights about how it modifies metabolome. To evaluate whether LGG affects the metabolite output of pathobionts, we inoculated LGG into gnotobiotic mice along with a gut-disruptive consortium containing Propionibacterium acnes, Turicibacter sanguinis, and Staphylococcus aureus (PTS).
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