Project description:Trained immunity is the phenomenon whereby innate immune cells such as monocytes or macrophages undergo functional reprogramming after exposure to certain microbial components, altering their responses to future exposures. Since trained immunity was first discovered, and has been commonly studied with the BCG vaccine, studies confirming trained immunity have often focused on human PBMCs. Thus, to study the mechanism of the vaccine in a well-established trained immunity model, we used human macrophages differentiated from monocytes isolated from PBMCs. The macrophages were expose to A + M + MA for 24 h, and rested two or five days prior to exposure to bacteria, as an ex vivo model with a five-day rest period has been used in numerous studies to confirm trained immunity.
Project description:Trained immunity is the phenomenon whereby innate immune cells such as monocytes or macrophages undergo functional reprogramming after exposure to certain microbial components, altering their responses to future exposures. Since trained immunity was first discovered, and has been commonly studied with the BCG vaccine, studies confirming trained immunity have often focused on human PBMCs. Thus, to study the mechanism of the vaccine in a well-established trained immunity model, we used human macrophages differentiated from monocytes isolated from PBMCs. The macrophages were expose to A + M + MA for 24 h, and rested two or five days prior to exposure to bacteria, as an ex vivo model with a five-day rest period has been used in numerous studies to confirm trained immunity.
Project description:Transcriptomic analysis of the temporal changes induced in mouse bone marrow derived macrophages (BMDMs) by the cytokine Interferon-beta over a timecourse of 0 to 24 hours of treatment. We set out to study the transcriptional events in mouse macrophages over time following stimulation with Interferon-beta. Mouse bone marrow derived macrophages were stimulated for 1, 2, 4, 8 and 24 hours with 10U/mL mouse interferon-beta or left untreated.
Project description:Kupffer cells are the first line of defense in the liver against pathogens, yet several microbes successfully target the liver, bypass immune surveillance, and effectively develop in this tissue. Our current, albeit poor, understanding of Kupffer cell-pathogen interactions has been largely achieved through the study of primary cells, requiring isolation from a large numbers of animals. To facilitate the study of Kupffer cell biology, an immortalized rat Kupffer cell line, RKC1, was developed. We performed a comparative global proteomic analysis of RKC1 and primary rat Kupffer cells (PRKC) to characterize their respective responses to lipopolysaccharide (LPS)-mediated immune stimulation. We identified patent differences in the proteomic response profile of RKC1 and PRKC to LPS. We observed that PRKC upregulated more immune function pathways and exhibited marked changes in cellular morphology following stimulation. We consequently analyzed the cytoskeletal signaling pathways of these cells in light of the fact that macrophages are known to induce cytoskeletal changes in response to pathogens. Our findings suggest that Kupffer cells respond differently to inflammatory stimulus than do monocyte-derived macrophages, and such data may provide insight into how pathogens, such as the malaria parasite, may have evolved mechanisms of liver entry through Kupffer cells without detection.
Project description:This study evaluated transcriptomic changes in monocyte-derived macrophages from healthy donors after stimulation with recombinant SLAMF7 or anti-SLAMF7 antibody, compared to macrophages without additional stimulation.
Project description:Kupffer cells are the first line of defense in the liver against pathogens, yet several microbes successfully target the liver, bypass immune surveillance, and effectively develop in this tissue. Our current, albeit poor, understanding of Kupffer cell-pathogen interactions has been largely achieved through the study of primary cells, requiring isolation from a large numbers of animals. To facilitate the study of Kupffer cell biology, an immortalized rat Kupffer cell line, RKC1, was developed. We performed a comparative global proteomic analysis of RKC1 and primary rat Kupffer cells (PRKC) to characterize their respective responses to lipopolysaccharide (LPS)-mediated immune stimulation. We identified patent differences in the proteomic response profile of RKC1 and PRKC to LPS. We observed that PRKC upregulated more immune function pathways and exhibited marked changes in cellular morphology following stimulation. We consequently analyzed the cytoskeletal signaling pathways of these cells in light of the fact that macrophages are known to induce cytoskeletal changes in response to pathogens. Our findings suggest that Kupffer cells respond differently to inflammatory stimulus than do monocyte-derived macrophages, and such data may provide insight into how pathogens, such as the malaria parasite, may have evolved mechanisms of liver entry through Kupffer cells without detection.