Project description:Our goal is to discriminate specific genes in live M.leprae-infected peritoneal macrophages in comparison to heat-killed M.leprae infected peritoneal macrophages using microarray. Two-condition experiment, Heat-killed M.leprae infected macrophage vs. Live M.leprae infected macrophage. Biological replicates: 16 mice (control), 8 heat-killed M. leprae mice (sample 1), and 8 live M. leprae infected mice (sample 2). Independently grown and harvested from isolator. One replicate per array.
Project description:Our goal is to discriminate specific genes in live M.leprae-infected peritoneal macrophages in comparison to heat-killed M.leprae infected peritoneal macrophages using microarray.
Project description:Bone marrow-derived dendritic cells were infected with live or heat-killed Bordetella and cells were analyzed on day 4 post-infection cells were >90% CD11c-positive
Project description:Background: Tuberculosis (TB) remains a major public health problem, especially in developing countries, with 1.5 million deaths annually worldwide. Macrophages (Mφs) are central to TB pathogenesis. Mφs play a key role in the outcome of the infection. More importantly, Mφs are the primary cell target of MTB, which has developed different strategies to multiply inside the Mφ phagosome. Here we aim to compare the transcriptome of Mycobacterium tuberculosis (MTB)-infected Mφs with that of cells stimulated with heat-killed MTB. Results: Here, we used live and heat-inactivated MTB to examine the impact of infection on the human macrophage response. Gene expression profiling has revealed about 3,259 genes differentially expressed in macrophages incubated with heat-inactivated MTB and 4,179 genes differentially expressed with live MTB. We found that the response to live and heat-inactivated MTB was strongly correlated (r>0.78). Conclusions: Our results show that macrophages respond in a similar manner to live and heat-inactivated MTB. We also identified some genes differentially expressed only in one condition.
Project description:In an attempt to understand M. leprae interaction with the human host, Stanford Genomics HEEBO Arrays were use to identify modulated genes in primary human Schwann cells infected with live M. leprae at a MOI of 100:1 for 24 hours. Dual channel competitive hybridizations between M. leprae infected and non infected Schwann cells including 3 independent biological replicates and a technical replicate in the form of dye swap.
Project description:Growing evidence suggests the importance of lipid metabolism in pathogenesis of tuberculosis. Neutral lipids form the majority of lipids in caseous granulomas characteristic of tuberculosis. Macrophage lipid droplets form the store house of these lipids, yet infection induced changes in the proteome of these dynamic organelles remains elusive. Here we employed quantitative proteomics to identify alterations induced upon infection with live Mycobacterium tuberculosis in comparison with heat killed bacilli and uninfected macrophages. Association of specific proteins coupled with lysosomal function was found to be increased upon infection with live M. tuberculosis. Biochemical and microscopy based evidence validated the enrichment of the small GTPase Arl8b, the guanine nucleotide effector LAMTOR1, and the scavenger receptor SCARB2 on lipid droplets during live Mtb infection. We validated the localization of Arl8b on lipid droplets using super-resolution microscopy. Increased abundance of these lysosomal proteins on lipid droplets upon infection with live Mtb suggests active manipulation of the host lipid droplets by the pathogen. Furthermore, depletion of lipid droplets upon stable knockdown of diacylglycerol O-acyltransferase led to reduction in lysosomal acidification of infected cells, suggesting an active role of lipid droplets in lysosomal function during infection.
Project description:In an attempt to understand M. leprae interaction with the human host, Stanford Genomics HEEBO Arrays were use to identify modulated genes in primary human Schwann cells infected with live M. leprae at a MOI of 100:1 for 24 hours.
Project description:Bovine tuberculosis, caused by Mycobacterium bovis, is a disease of considerable economic importance yet comparatively little is known about the bovine immune response to the disease. Alveolar macrophages are one of the first cells to encounter mycobacteria following infection. In this experiment we investigated the early transcriptional response of bovine alveolar macrophages following infection with M. bovis. The transcriptional response to heat-killed M. bovis was also investigated to look for genes that are only differentially transcribed in response to the live organism. Five-condition experiment, uninfected, live and heat-killed M. bovis-infected bovine alveolar macrophages from five cattle infected for two and four hours. Comparisons were within animal. Dye swaps were incorporated into the design.
Project description:The effects of Candida albicans on the metastatic activity of oral squamous cell carcinoma was observed in vitro and in vivo. In the in vitro experimental setup HO-1-N-1 and HSC-2 human oral squamous cell carcinoma cell lines were treated with zymosan, heat-killed Candida albicans, heat-killed C. parapsilosis, live C. albicans and live C. parapsilosis. Whole transcriptomics was performed of the human tumor cells. In the in vivo experiment human HSC-2 tumor cells were injected to the tongue of mice. Whole transcriptomic analysis was performed of the human HSC-2 derived tumor cells comparing control tumor and oral candidiasis treated tumor.
Project description:The goal was to examine gene expression after in vivo activation by different vaccine strains of Listeria monocytogenes. Heat killed LM, irradiated LM and live actA- LM were used to immunize mice after transfer of OT-I cells
