Project description:SiglecF positive bone marrow leukocytes were submitted to SCRNAseq in order to investigate eosinophil development and amplification during eosinophilia.
Project description:Diesel exhaust particles (DEPs) are the main component of traffic-related air pollution and implicated in asthma pathogenesis and exacerbation. However, the mechanistic link between DEP exposure and worsening asthma symptoms remains unclear. Here, we characterized a unique subset of neutrophils that express a high level of Sialic acid-binding, Ig-like lectin (Siglec)-F. DEP exposure instructs SiglecF expression on neutrophils by increasing P2X1 receptors sensing damage-associated molecular patterns (DAMPs) molecules, ATP. SiglecFhigh neutrophils stimulated by DEP release neutrophil extracellular traps (NETs) to cause tissue damage and IL-17A production, and potentiated type 2 cytokine production by producing cysteinyl leukotrienes. As a result, SiglecFhigh neutrophils worsen airway hyperresponsiveness (AHR) as well as airway inflammations. We also confirmed that siglec8, corresponding to murine SiglecF, expression increased in neutrophils from patients with asthma, and they correlated with the severity of the disease. Together, these translational findings suggest a novel SiglecF expressing neutrophils in the development of severe asthma by linking type 2 and type 3 responses.
Project description:Parallel DNA and RNA profiling of EpCAM-positive cells in bone marrow and primary tumor tissue with positive disseminated tumor cell (DTC) count via immunomagnetic Enrichment/Flow Cytometry (IE/FC) of metastatic breast cancer (MBC) patients confirm their malignant nature We developed a novel approach to isolate tumor cells with high purity from bone marrow which was subjected to immunomagnetic enrichment using EpCAM beads followed by fluorescence activated cell sorting (IE/FACS) to isolate EpCAM-positive cells away from leukocytes (CD45+). For DNA profiling, sorted cells were subjected to BAC array comparative genomic hybridization analysis following whole genome amplification. For RNA profiling, QPCR analysis was performed on sixty four (64) cancer-related genes using Taqman® low density arrays. For non-tumor controls, RNA profiling was performed on matched leukocytes (CD45+) isolated from the same enriched bone marrow samples.
Project description:We developed a novel approach to isolate tumor cells with high purity from bone marrow which was subjected to immunomagnetic enrichment using EpCAM beads followed by fluorescence activated cell sorting (IE/FACS) to isolate EpCAM-positive cells away from leukocytes (CD45+). For RNA profiling, QPCR analysis was performed on sixty four (64) cancer-related genes using Taqman® low density arrays. For non-tumor controls, RNA profiling was performed on matched leukocytes (CD45+) isolated from the same enriched bone marrow samples from 17 of the 30 patients.
Project description:Genome-wide transcriptional profiling results were used to examine the effect of a psychological stress paradigm for rodents (chronic restraint) on indicies of biological aging in murine femoral bone marrow leukocytes and to assess the extent to which propranolol could block such effects.
Project description:We identified the genes regulated by hindlimb ischemia in leukocytes and hematopoietic stem and progenitor cells isolated from the bone marrow in mice. We selected some of these genes based on their expression and further evaluated their functions using a wise array of techniques.
Project description:Cell suspensions were prepared from lungs dissociated at baseline, at 14 days, and at 28 days after injury. Cells (siglecF+CD11C+ and MHCII-high or -low) were isolated with FACS. RNA-seq was performed with sorted cells.
Project description:Respiratory viral infections trigger a robust inflammatory response in the lung, producing cytokines, chemokines, and growth factors that promote infiltration of effector leukocytes. Whereas the role of chemokines and infiltrating leukocytes in antiviral immunity is well studied, the effect that lung cytokines have on leukocytes in distal hematopoietic and lymphoid tissues and their role in antiviral immunity is unknown. We show that, during infection with influenza or Sendai virus, the lung communicates with the sterile bone marrow, the primary site of hematopoiesis, through type I interferons. While in the bone marrow, leukocytes exposed to type I interferons activate an antiviral transcriptional program and become resistant to infection with different viruses. The protected bone marrow leukocytes are capable of migrating to the infected lung and contribute to virus clearance. These findings show that appropriate instruction of cells during their development in the bone marrow is needed for effective control of infection.